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CLAIMS 1. A compound of Formula (CY) (CY), or a pharmaceutically acceptable salt thereof, wherein: R1 is selected from the group consisting of -OH, -OAc, R1a, ; Z1 is optionally substituted C1-C6 alkyl; X1 is optionally substituted C2-C6 alkylenyl; X2 is selected from the group consisting of a bond, -CH2- and -CH2CH2-; X2’ is selected from the group consisting of a bond, -CH2- and -CH2CH2-; X3 is selected from the group consisting of a bond, -CH2- and -CH2CH2-; X3’ is selected from the group consisting of a bond, -CH2- and -CH2CH2-; X4 and X5 are independently optionally substituted C2-C14 alkylenyl or optionally substituted C2-C14 alkenylenyl; Y1 and Y2 are independently selected from the group consisting of , wherein the bond marked with an "*" is attached to X4 or X5; each Z2 is independently H or optionally substituted C1-C8 alkyl; each Z3 is indpendently optionally substituted C1-C6 alkylenyl; R2 is selected from the group consisting of optionally substituted C4-C20 alkyl, optionally substituted C2-C14 alkenyl, and –(CH2)pCH(OR6)(OR7); R3 is selected from the group consisting of optionally substituted C4-C20 alkyl, optionally substituted C2-C14 alkenyl, or -(CH2)qCH(OR8)(OR9); R1a is: ; R2a, R2b, and R2c are independently hydrogen and C1-C6 alkyl; R3a, R3b, and R3c are independently hydrogen and C1-C6 alkyl; R4a, R4b, and R4c are independently hydrogen and C1-C6 alkyl; R5a, R5b, and R5c are independently hydrogen and C1-C6 alkyl; R6, R7, R8, and R9 are independently optionally substituted C1-C14 alkyl, optionally substituted C2-C14 alkenyl, or -(CH2)m-A-(CH2)nH; each A is indepenently a C3-C8 cycloalkylenyl; each m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; p is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, and 7; and q is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, and 7. 2. The compound of claim 1, wherein the compound is of Formula (CY-I'), (CY-II), (CY- III), (CY-IV), or (CY-V): (CY-IV), and (CY-V), or a pharmaceutically acceptable salt thereof, wherein X2 and X3 are independently a bond, -CH2-, or -CH2CH2-; and X6 and X7 are independently -CH2- or -CH2CH2-. 3. The compound of claim 1 or 2, wherein R1 is selected from the group consisting of -OH, . 4. The compound of claim 1 or 2, wherein R1 is -OH. 5. The compound of any of claims 1-4, wherein Y1 and Y2 are independently: . 6. The compound of any of claims 1-5, wherein the compound is of Formula (CY-VI) or (CY-VII): (CY-VI) (CY-VII), or pharmaceutically acceptable salt thereof. 7. The compound of any of claims 1-5, wherein the compound is of Formula (CY-VIII) or (CY-IX): (CY- VIII) (CY- IX), or pharmaceutically acceptable salt thereof. 8. The compound of any of claims 1-5, wherein the compound is of Formula (CY-IV-a), (CY-IV-b), or (CY-IV-c) (CY-IV-a) (CY-IV-b) (CY-IV-c) ), or pharmaceutically acceptable salt thereof. 9. The compound of any of claims 1-5, wherein the compound is of Formula (CY-IV-d), (CY-IV-e), or (CY-IV-f) (CY-IV-d) (CY-IV-e) (CY-IV-f) ), or pharmaceutically acceptable salt thereof. 10. The compound of any of claims 1-9, wherein X4 is optionally substituted C2-C10 alkylenyl and X5 is optionally substituted C2-C10 alkylenyl. 11. The compound of any of claims 1-9, wherein X4 is optionally substituted C2-C6 alkylenyl and X5 is optionally substituted C2-C6 alkylenyl. 12. The compound of claim 11, wherein X4 is optionally substituted C2 alkylenyl and X5 is optionally substituted C2 alkylenyl. 13. The compound of claim 11, wherein X4 is optionally substituted C2 alkylenyl and X5 is optionally substituted C2 alkylenyl. 14. The compound of any of claims 1-13, wherein X1 is optionally substituted C2-C4 alkylenyl. 15. The compound of any of claims 1-13, wherein X1 is optionally substituted C2 alkylenyl. 16. The compound of any of claims 1-15, wherein R2 is –(CH2)pCH(OR6)(OR7). 17. The compound of claim 16, wherein p is selected from the group consisting of 0, 1, and 2. 18. The compound of claim 16, wherein p is 0. 19. The compound of any of claims 1-18, wherein R3 is -(CH2)qCH(OR8)(OR9). 20. The compound of claim 19, wherein q is selected from the group consisting of 0, 1, and 2. 21. The compound of claim 19, wherein q is 0. 22. The compound of any of claims 1-15 or 19-21, wherein R2 is optionally substituted C4- C20 alkyl. 23. The compound of claim 22, wherein R2 is optionally substituted C10-C20 alkyl. 24. The compound of claim 22, wherein R2 is optionally substituted C10-C15 alkyl. 25. The compound of claim 22, wherein R2 is optionally substituted C15-C20 alkyl. 26. The compound of any of claims 1-18, wherein R3 is optionally substituted C4-C20 alkyl. 27. The compound of claim 26, wherein R3 is optionally substituted C10-C20 alkyl. 28. The compound of claim 26, wherein R3 is optionally substituted C10-C15 alkyl. 29. The compound of claim 26, wherein R3 is optionally substituted C15-C20 alkyl. 30. The compound of any of claims 1-21, wherein R6 is optionally substituted C1-C14 alkyl; R7 is optionally substituted C1-C14 alkyl; R8 is optionally substituted C1-C14 alkyl; and R9 is optionally substituted C1-C14 alkyl. 31. The compound of any of claim 30, wherein R6 is optionally substituted C3-C10 alkyl; R7 is optionally substituted C3-C10 alkyl; R8 is optionally substituted C3-C10 alkyl; and R9 is optionally substituted C3-C10 alkyl. 32. The compound of any of claim 30, wherein R6 is optionally substituted C6-C10 alkyl; R7 is optionally substituted C6-C10 alkyl; R8 is optionally substituted C6-C10 alkyl; and R9 is optionally substituted C6-C10 alkyl. 33. The compound of any of claims 1-15, wherein R2 is selected from the group consisting of 34. The compound of any of claims 1-15 or 33, wherein R3 is selected from the group consisting of 35. A compound selected from the group consisting of or a pharmaceutically acceptable salt thereof. 36. A pharmaceutical composition comprising: a) a polynucleotide, and b) a delivery vehicle comprising the compound of any of claims 1-35. 37. The pharmaceutical composition of claim 36, wherein the polynucleotides are DNA. 38. The pharmaceutical composition of claim 37, wherein the polynucleotides are RNA. 39. The pharmaceutical composition of claim 38, wherein the RNA are short interfering RNA (siRNA). 40. The pharmaceutical composition of claim 39, wherein the siRNA inhibits or suppresses the expression of a target of interest in a cell. 41. The pharmaceutical composition of any one of claims 36-39, wherein the polynucleotide comprises at least one modification. 42. The pharmaceutical composition of any of claims 36-41, further comprises an additional cationic lipid. 43. The pharmaceutical composition of any of claims 36-42, further comprising a neutral lipid. 44. The pharmaceutical composition of any of claims 36-43, further comprising an anionic lipid. 45. The pharmaceutical composition of any of claims 36-44, further comprises a helper lipid. 46. The pharmaceutical composition of any of claims 36-45, further comprises a stealth lipid. 47. The pharmaceutical composition of any of claims 36-46, wherein the weight ratio of the lipids and the polynucleotide is from about 100:1 to about 1:1. 48. A vaccine formulation comprising the pharmaceutical composition of any of claims 36- 47. 49. A vaccine preparation comprising the pharmaceutical composition of any of claims 36- 47. 50. A method of vaccinating a subject against an infectious agent comprising: c) contacting a subject with the vaccine formulation of claim 48 or the vaccine preparation of claim 49, and d) eliciting an immune response. 51. A method of delivering a polynucleotide encoding at least one protein of interest to an immune cell of a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of any one of claims 36-47. 52. The method of claim 51, wherein the immune cell is a T cell. 53. The method of claim 52, wherein the T cell is a CD8+ T cell. 54. The method of claim 52, wherein the T cell is a T regulatory cell. 55. The method of claim 52, wherein the T cell is CD4+ T cell. 56. The method of claim 51, wherein the immune cell is a macrophage, dendritic cell, or liver immune cell. 57. A lipid nanoparticle (LNP) comprising a compound of any one of claims 1-35, or a pharmaceutically acceptable salt thereof. 58. The LNP of claim 57, further comprising: (a) a PEG-lipid; (b) a structural lipid; and (c) a non-ionizable lipid and/or a zwitterionic lipid. 59. The LNP of claim 58, wherein the ionizable lipid comprises an ionizable amino lipid. 60. The LNP of any one of claims 58-59, wherein the PEG-lipid is selected from the group consisting of PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, and PEG-DSPE. 61.. The LNP of any one of claims 58-60, wherein the structural lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, an alpha-tocopherol. 62. The LNP of any one of claims 58-61, wherein the non-ionizable lipid is a phospholipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn- glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1.2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1- palmitoyl-2-oleoyl-sn-glycero-3-phosphocho line (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuc cinoyl-sn- glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero- 3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2- diphytanoylsn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero- 3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2- dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3- phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2- dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sodium (S)-2- ammonio-3-((((R)-2-(oleoyloxy)-3- (stearoyloxy)propoxy)oxidophosphoryl)oxy)propanoate (L-α-phosphatidylserine; Brain PS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleoyl- phosphatidylethanolamine4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE- mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L- serine) (DOPS), acell-fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl-phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), 1,2-dioleoyl-sn-glycero-3-phosphate (18:1 PA; DOPA), ammonium bis((S)-2- hydroxy-3-(oleoyloxy)propyl) phosphate (18:1 DMP; LBPA), 1,2-dioleoyl-sn-glycero-3- phospho-(1’-myo-inositol) (DOPI; 18:1 PI), 1,2-distearoyl-sn-glycero-3-phospho-L- serine (18:0 PS), 1,2-dilinoleoyl-sn-glycero-3-phospho-L-serine (18:2 PS), 1-palmitoyl-2- oleoyl-sn-glycero-3-phospho-L-serine (16:0-18:1 PS; POPS), 1-stearoyl-2-oleoyl-sn- glycero-3-phospho-L-serine (18:0-18:1 PS), 1-stearoyl-2-linoleoyl-sn-glycero-3-phospho- L-serine (18:0-18:2 PS), 1-oleoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:1 Lyso PS), 1-stearoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:0 Lyso PS), and sphingomyelin. 63. The LNP of any one of claims 58-62, further comprising a targeting moiety. 64. The LNP of claim 63, wherein the targeting moiety is an antibody or a fragment thereof. 65. The LNP of any one of claims 58-64, further comprising an active agent. 66. The LNP of claim 65, wherein the active agent is a nucleic acid. 67. The LNP of claim 66, wherein the nucleic acid is a ribonucleic acid. 68. The LNP of claim 67, wherein the ribonucleic acid is at least one ribonucleic acid selected from the group consisting of a small interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a microRNA (miRNA), a Dicer-substrate RNA (dsRNA), a small hairpin RNA (shRNA), a messenger RNA (mRNA), and a long non-coding RNA (lncRNA). 69. The LNP of claim 66, wherein the nucleic acid is a messenger RNA (mRNA) or a circular RNA. 70. The LNP of claim 69, wherein the mRNA includes an open reading frame encoding a cancer antigen. 71. The LNP of claim 69, wherein the mRNA includes an open reading frame encoding an immune checkpoint modulator. 72. The LNP of any one of claims 69-71, wherein the mRNA includes at least one motif selected from the group consisting of a stem loop, a chain terminating nucleoside, a polyA sequence, a polyadenylation signal, and a 5' cap structure. 73. The LNP of claim 66, wherein, wherein the nucleic acid is a polynucleotide that encodes a protein selected from SEQ ID NOs: 1-54. 74. The LNP of claim 66, wherein the nucleic acid is suitable for a genome editing technique. 75. The LNP of claim 74, wherein the genome editing technique is clustered regularly interspaced short palindromic repeats (CRISPR) or transcription activator-like effector nuclease (TALEN). 76. The LNP of claim 66, wherein the nucleic acid is at least one nucleic acid suitable for a genome editing technique selected from the group consisting of a CRISPR RNA (crRNA), a trans-activating crRNA (tracrRNA), a single guide RNA (sgRNA), and a DNA repair template. 77. The LNP of any one of claims 69, wherein the mRNA is at least 30 nucleotides in length. 78. The LNP of any one of claims 69, wherein the mRNA is at least 300 nucleotides in length. 79. A pharmaceutical composition comprising an LNP of any one of claims 57-78, and a pharmaceutically acceptable carrier. 80. The pharmaceutical composition of claim 79, formulated for intravenous or intramuscular administration. 81. The pharmaceutical composition of claim 80, which is formulated for intravenous administration. 82. A method for delivering a nucleic acid to a cell comprising contacting the cell with an LNP of any one of claims 57-78 or a pharmaceutical composition of any of claims 79-82. 83. A method for treating a disease characterized by a deficiency of a functional protein, the method comprising administering to a subject having the disease, an LNP formulation comprising an LNP of any one of claims 82, wherein the mRNA encodes the functional protein or a protein having the same biological activity as the functional protein. 84. A method for treating a disease characterized by overexpression of a polypeptide, comprising administering to a subject having the disease an LNP formulation comprising an LNP of any one of claims 57-78 and an siRNA, wherein the siRNA targets expression of the overexpressed polypeptide. |
R 3 [0501] In some embodiments, R 3 is selected from the group consisting of optionally substituted C 4 -C 20 alkyl, optionally substituted C 2 -C 14 alkenyl, and –(CH 2 ) q CH(OR 6 )(OR 7 ). [0502] In some embodiments, R 3 is optionally substituted C4-C20 alkyl. In some embodiments, R 3 is optionally substituted C 8 -C 17 alkyl. In some embodiments, R 3 is optionally substituted C9-C16 alkyl. In some embodiments, R 3 is optionally substituted C8-C10 alkyl. In some embodiments, R 3 is optionally substituted C 11 -C 13 alkyl. In some embodiments, R 3 is optionally substituted C14-C16 alkyl. In some embodiments, R 3 is optionally substituted C9 alkyl. In some embodiments, R 3 is optionally substituted C10 alkyl. In some embodiments, R 3 is optionally substituted C 11 alkyl. In some embodiments, R 3 is optionally substituted C 12 alkyl. In some embodiments, R 3 is optionally substituted C13 alkyl. In some embodiments, R 3 is optionally substituted C 14 alkyl. In some embodiments, R 3 is optionally substituted C 15 alkyl. In some embodiments, R 3 is optionally substituted C16 alkyl. [0503] In some embodiments, R 3 is optionally substituted C 2 -C 14 alkenyl. In some embodiments, R 3 is optionally substituted C5-C14 alkenyl. In some embodiments, R 3 is optionally substituted C 7 -C 14 alkenyl. In some embodiments, R 3 is optionally substituted C 9 - C14 alkenyl. In some embodiments, R 3 is optionally substituted C10-C14 alkenyl. In some embodiments, R 3 is optionally substituted C 12 -C 14 alkenyl. [0504] In some embodiments, R 3 is -(CH2)qCH(OR 8 )(OR 9 ). In some embodiments, R 3 is -CH(OR 8 )(OR 9 ). In some embodiments, R 3 is -CH 2 CH(OR 8 )(OR 9 ). In some embodiments, R 3 is -(CH2)2CH(OR 8 )(OR 9 ). In some embodiments, R 3 is -(CH2)3CH(OR 8 )(OR 9 ). In some embodiments, R 3 is -(CH 2 ) 4 CH(OR 8 )(OR 9 ). [0505] In some embodiments, R 3 is selected from the group consisting of
R 6 , R 7 , R 8 , R 9 [0506] In some embodiments, R 6 , R 7 , R 8 , and R 9 are independently optionally substituted C1- C 14 alkyl, optionally substituted C 2 -C 14 alkenyl, or -(CH 2 ) m -A-(CH 2 ) n H. In some embodiments, R 6 , R 7 , R 8 , and R 9 are independently optionally substituted C1-C14 alkyl. In some embodiments, R 6 , R 7 , R 8 , and R 9 are independently optionally substituted C 2 -C 14 alkenyl. In some embodiments, R 6 , R 7 , R 8 , and R 9 are independently -(CH2)m-A-(CH2)nH. [0507] In some embodiments, R 6 is optionally substituted C 1 -C 14 alkyl, optionally substituted C2-C14 alkenyl, or -(CH2)m-A-(CH2)nH. In some embodiments, R 6 is optionally substituted C3- C10 alkyl. In some embodiments, R 6 is optionally substituted C4-C10 alkyl. In some embodiments, R 6 is independently optionally substituted C 5 -C 10 alkyl. In some embodiments, R 6 is optionally substituted C9-C10 alkyl. In some embodiments, R 6 is optionally substituted C 1 -C 14 alkyl. In some embodiments, R 6 is optionally substituted C 2 -C 14 alkenyl. In some embodiments, R 6 is –(CH2)m-A-(CH2)nH. [0508] In some embodiments, R 7 is optionally substituted C 1 -C 14 alkyl, optionally substituted C2-C14 alkenyl, or –(CH2)m-A-(CH2)nH. In some embodiments, R 7 is optionally substituted C3- C 10 alkyl. In some embodiments, R 7 is optionally substituted C 4 -C 10 alkyl. In some embodiments, R 7 is optionally substituted C5-C10 alkyl. In some embodiments, R 7 is optionally substituted C 9 -C 10 alkyl. In some embodiments, R 7 is optionally substituted C 1 -C 14 alkyl. In some embodiments, R 7 is optionally substituted optionally substituted C2-C14 alkenyl. In some embodiments, R 7 is –(CH 2 ) m -A-(CH 2 ) n H. [0509] In some embodiments, R 8 is optionally substituted C1-C14 alkyl, optionally substituted C 2 -C 14 alkenyl, or –(CH 2 ) m -A-(CH 2 ) n H. In some embodiments, R 8 is optionally substituted C 3 - C10 alkyl. In some embodiments, R 8 is optionally substituted C4-C10 alkyl. In some embodiments, R 8 is optionally substituted C 5 -C 10 alkyl. In some embodiments, R 8 is optionally substituted C9-C10 alkyl. In some embodiments, R 8 is optionally substituted C1-C14 alkyl. In some embodiments, R 8 is optionally substituted C2-C14 alkenyl. In some embodiments, R 8 is –(CH 2 ) m -A-(CH 2 ) n H. [0510] In some embodiments, R 9 is optionally substituted C1-C14 alkyl, optionally substituted C 2 -C 14 alkenyl, or –(CH 2 ) m -A-(CH 2 ) n H. In some embodiments, R 9 is optionally substituted C 3 - C10 alkyl. In some embodiments, R 9 is optionally substituted C4-C10 alkyl. In some embodiments, R 9 is optionally substituted C 5 -C 10 alkyl. In some embodiments, R 9 is optionally substituted C9-C10 alkyl. In some embodiments, R 9 is optionally substituted C1-C14 alkyl. In some embodiments, R 9 is optionally substituted C 2 -C 14 alkenyl. In some embodiments, R 9 is – (CH2)m-A-(CH2)nH. [0511] In some embodiments, each m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, each m is 0. In some embodiments, each m is 1. In some embodiments, each m is 2. In some embodiments, each m is 3. In some embodiments, each m is 4. In some embodiments, each m is 5. In some embodiments, each m is 6. In some embodiments, each m is 7. In some embodiments, each m is 8. In some embodiments, each m is 9. In some embodiments, each m is 10. In some embodiments, each m is 11. In some embodiments, each m is 12. [0512] In some embodiments, each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In some embodiments, each n is 0. In some embodiments, each n is 1. In some embodiments, each n is 2. In some embodiments, each n is 3. In some embodiments, each n is 4. In some embodiments, each n is 5. In some embodiments, each n is 6. In some embodiments, each n is 7. In some embodiments, each n is 8. In some embodiments, each n is 9. In some embodiments, each n is 10. In some embodiments, each n is 11. In some embodiments, each n is 12. [0513] In some embodiments, each A is independently a C3-C8 cycloalkylenyl. In some embodiments, each A is cyclopropylenyl. X 1 [0514] In some embodiments, X 1 is optionally substituted C 2 -C 6 alkylenyl. In some embodiments, X 1 is optionally substituted C2-C5 alkylenyl. In some embodiments, X 1 is optionally substituted C 2 -C 4 alkylenyl. In some embodiments, X 1 is optionally substituted C 2 - C3 alkylenyl. In some embodiments, X 1 is optionally substituted C2 alkylenyl. In some embodiments, X 1 is optionally substituted C 3 alkylenyl. In some embodiments, X 1 is optionally substituted C4 alkylenyl. In some embodiments, X 1 is optionally substituted C5 alkylenyl. In some embodiments, X 1 is optionally substituted C 6 alkylenyl. In some embodiments, X 1 is optionally substituted –(CH2)2-. In some embodiments, X 1 is optionally substituted –(CH2)3-. In some embodiments, X 1 is optionally substituted –(CH 2 ) 4 -. In some embodiments, X 1 is optionally substituted –(CH2)5-. In some embodiments, X 1 is optionally substituted –(CH2)6-. X 2 [0515] In some embodiments, X 2 is selected from the group consisting of a bond, -CH 2 - and - CH2CH2-. In some embodiments, X 2 is a bond. In some embodiments, X 2 is -CH2-. In some embodiments, X 2 is -CH 2 CH 2 -. X 2’ [0516] In some embodiments, X 2’ is selected from the group consisting of a bond, -CH 2 - and - CH2CH2-. In some embodiments, X 2’ is a bond. In some embodiments, X 2’ is -CH2-. In some embodiments, X 2’ is -CH 2 CH 2 -. X 3 [0517] In some embodiments, X 3 is selected from the group consisting of a bond, -CH2- and - CH2CH2-. In some embodiments, X 3 is a bond. In some embodiments, X 3 is -CH2-. In some embodiments, X 3 is -CH 2 CH 2 -. X 3’ [0518] In some embodiments, X 3’ is selected from the group consisting of a bond, -CH2- and - CH 2 CH 2 -. In some embodiments, X 3’ is a bond. In some embodiments, X 3’ is -CH 2 -. In some embodiments, X 3’ is -CH2CH2-. X 4 [0519] In some embodiments, X 4 is selected from the group consting of optionally substituted C 2 -C 14 alkylenyl and optionally substituted C 2 -C 14 alkenylenyl. In some embodiments, X 4 is optionally substituted C2-C14 alkylenyl. In some embodiments, X 4 is optionally substituted C2- C 10 alkylenyl. In some embodiments, X 4 is optionally substituted C 2 -C 8 alkylenyl. In some embodiments, X 4 is optionally substituted C2-C6 alkylenyl. In some embodiments, X 4 is optionally substituted C 3 -C 6 alkylenyl. In some embodiments, X 4 is optionally substituted C 3 alkylenyl. In some embodiments, X 4 is optionally substituted C4 alkylenyl. In some embodiments, X 4 is optionally substituted C 5 alkylenyl. In some embodiments, X 4 is optionally substituted C6 alkylenyl. In some embodiments, X 4 is optionally substituted –(CH2)2-. In some embodiments, X 4 is optionally substituted –(CH 2 ) 3 -. In some embodiments, X 4 is optionally substituted –(CH2)4-. In some embodiments, X 4 is optionally substituted –(CH2)5-. In some embodiments, X 4 is optionally substituted –(CH 2 ) 6 -. 5 [0520] In some embodiments, X 5 is selected from the group consting of optionally substituted C 2 -C 14 alkylenyl and optionally substituted C 2 -C 14 alkenylenyl. In some embodiments, X 5 is optionally substituted C2-C14 alkylenyl. In some embodiments, X 5 is optionally substituted C2- C 10 alkylenyl. In some embodiments, X 5 is optionally substituted C 2 -C 8 alkylenyl. In some embodiments, X 5 is optionally substituted C2-C6 alkylenyl. In some embodiments, X 5 is optionally substituted C3-C6 alkylenyl. In some embodiments, X 5 is optionally substituted C3 alkylenyl. In some embodiments, X 5 is optionally substituted C 4 alkylenyl. In some embodiments, X 5 is optionally substituted C5 alkylenyl. In some embodiments, X 5 is optionally substituted C 6 alkylenyl. In some embodiments, X 5 is optionally substituted –(CH 2 ) 2 -. In some embodiments, X 5 is optionally substituted –(CH2)3-. In some embodiments, X 5 is optionally substituted –(CH 2 ) 4 -. In some embodiments, X 5 is optionally substituted –(CH 2 ) 5 -. In some embodiments, X 5 is optionally substituted –(CH2)6-. Y 1 [0521] In some embodiments, Y 1 is selected from the group consisting of , [0522] In some embodiments, Y 1 is selected from the group consisting of . [0523] In some embodiments, Y 1 is . [0524] In some embodiments, Y 1 is . [0525] In some embodiments, Y 1 is . [0527] In some embodiments, Y 2 is selected from the group consisting of , [0528] In some embodiments, Y 2 is selected from the group consisting of . [0529] In some embodiments, Y 2 is . [0530] In some embodiments, Y 2 is . [0531] In some embodiments, Y 2 is . [0532] In some embodiments, Y 2 is . Formula (CY-I’) [0533] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’): , or a pharmaceutically acceptable salt thereof, wherein: R 1 is -OH, R 1a , ; Z 1 is optionally substituted C1-C6 alkyl; X 1 is optionally substituted C 2 -C 6 alkylenyl; X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl or optionally substituted C2-C14 alkenylenyl; Y 1 and Y 2 are independently , wherein the bond marked with an "*" is attached to X 4 or X 5 ; each Z 2 is independently H or optionally substituted C 1 -C 8 alkyl; each Z 3 is indpendently optionally substituted C1-C6 alkylenyl; R 2 is optionally substituted C 4 -C 20 alkyl, optionally substituted C 2 -C 14 alkenyl, or -CH(OR 6 )(OR 7 ); R 3 is optionally substituted C4-C20 alkyl, optionally substituted C2-C14 alkenyl, or -CH(OR 8 )(OR 9 ); R 1a is: ; R 2a , R 2b , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C1-C6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C 1 -C 6 alkyl; R 5a , R 5b , and R 5c are independently hydrogen and C1-C6 alkyl; R 6 , R 7 , R 8 , and R 9 are independently optionally substituted C 1 -C 14 alkyl, optionally substituted C2-C14 alkenyl, or -(CH2)m-A-(CH2)nH; A is a C 3 -C 8 cycloalkylenyl; each m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. [0534] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’), wherein: R 1 is -OH, R 1a , , wherein Z 1 is optionally substituted C1-C6 alkyl; X 1 is optionally substituted C 2 -C 6 alkylenyl; X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; Y 1 and Y 2 are independently ; R 2 and R 3 are independently optionally substituted C4-C20 alkyl; R 1a is: ; R 2a , R 2b , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C 1 -C 6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C1-C6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C 1 -C 6 alkyl. [0535] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein: R 1 is -OH, R 1a , , wherein Z 1 is optionally substituted C1-C6 alkyl; X 1 is optionally substituted C 2 -C 6 alkylenyl; X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; Y 1 and Y 2 are independently ; R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl; R 1a is: ; R 2a , R 2b , and R 2c are independently hydrogen and C 1 -C 6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C1-C6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C 1 -C 6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C1-C6 alkyl. [0536] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’), wherein R 1 is -OH, . [0537] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’), wherein Y 1 and Y 2 are independently: . [0538] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’), wherein R 2 is -CH(OR 6 )(OR 7 ). [0539] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-I’), wherein R 3 is -CH(OR 8 )(OR 9 ). [0540] Non-limiting examples of lipids having a structure of Formula (CY-I’) include compounds CY1, CY2, CY3, CY9, CY10, CY11, CY12, CY22, CY23, CY24, CY30, CY31, CY32, CY33, CY43, CY44, CY45, CY50, CY51, CY52, and CY53. Formula (CY-II’) [0541] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’): , or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I’). [0542] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein: R 1 is -OH, R 1a , , wherein Z 1 is optionally substituted C1-C6 alkyl; X 1 is optionally substituted C 2 -C 6 alkylenyl; X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; Y 1 and Y 2 are independently ; R 2 and R 3 are independently optionally substituted C4-C20 alkyl; R 1a is: R 2a , R 2b , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C 1 -C 6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C1-C6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C 1 -C 6 alkyl. [0543] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein R 1 is -OH, . [0544] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein Y 1 and Y 2 are independently: . [0545] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein R 2 is -CH(OR 6 )(OR 7 ). [0546] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-II’), wherein R 3 is -CH(OR 8 )(OR 9 ). [0547] Non-limiting examples of lipids having a structure of Formula (CY-II’) include compounds CY4, CY5, CY16, CY17, CY18, CY25, CY26, CY37, CY38, CY39, CY46, CY56, and CY57. Formula (CY-III’) [0548] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’): or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I’). [0549] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’), wherein R 1 is -OH, R 1a , , wherein Z 1 is optionally substituted C1-C6 alkyl; X 1 is optionally substituted C 2 -C 6 alkylenyl; X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; Y 1 and Y 2 are independently ; R 2 and R 3 are independently optionally substituted C4-C20 alkyl; R 1a is: ; R 2a , R 2b , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C 1 -C 6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C1-C6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C 1 -C 6 alkyl. [0550] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’), wherein R 1 is -OH, . [0551] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’), wherein Y 1 and Y 2 are independently: . [0552] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’), wherein R 2 is -CH(OR 6 )(OR 7 ). [0553] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-III’), wherein R 3 is -CH(OR 8 )(OR 9 ). [0554] Non-limiting examples of lipids having a structure of Formula (CY-III’) include CY6, CY14, CY27, CY35, CY47, and CY55. Formula (CY-IV) [0555] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’): , or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I’). [0556] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’), wherein: R 1 is -OH, R 1a , wherein Z 1 is optionally substituted C 1 -C 6 alkyl; X 1 is optionally substituted C2-C6 alkylenyl; X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; Y 1 and Y 2 are independently R 2 and R 3 are independently optionally substituted C4-C20 alkyl; R 1a is: ; R 2a , R 2b , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C 1 -C 6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C1-C6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C 1 -C 6 alkyl [0557] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’), wherein R 1 is -OH, . [0558] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’), wherein Y 1 and Y 2 are independently: . [0559] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’), wherein R 2 is -CH(OR 6 )(OR 7 ). [0560] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-IV’), wherein R 3 is -CH(OR 8 )(OR 9 ). [0561] Non-limiting examples of lipids having a structure of Formula (CY-IV’) include compounds CY7, CY8, CY19, CY20, CY21, CY28, CY29, CY40, CY41, CY42, CY48, CY49, CY58, CY59, and CY60. Formula (CY-V’) [0562] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-V’): , or a pharmaceutically acceptable salt thereof, wherein: X 6 and X 7 are independently -CH 2 - or -CH 2 CH 2 -; and R 1 , R 2 , R 3 , X 1 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I’). [0563] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-V’), wherein: R 1 is -OH, R 1a , , wherein Z 1 is optionally substituted C 1 -C 6 alkyl; X 1 is optionally substituted C2-C6 alkylenyl; X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; Y 1 and Y 2 are independently ; R 2 and R 3 are independently optionally substituted C4-C20 alkyl; R 1a is: ; , and R 2c are independently hydrogen and C1-C6 alkyl; R 3a , R 3b , and R 3c are independently hydrogen and C 1 -C 6 alkyl; R 4a , R 4b , and R 4c are independently hydrogen and C1-C6 alkyl; and R 5a , R 5b , and R 5c are independently hydrogen and C 1 -C 6 alkyl [0564] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-V’), wherein Y 1 and Y 2 are independently: . [0565] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-V’), wherein R 2 is -CH(OR 6 )(OR 7 ). [0566] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-V’), wherein R 3 is -CH(OR 8 )(OR 9 ). [0567] Non-limiting examples of lipids having a structure of Formula (CY-V’) include compounds CY13, CY15, CY34, CY36, and CY54. [0568] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’): , or a pharmaceutically acceptable salt thereof, wherein R 1 , R 6 , R 7 , R 8 , R 9 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I’). [0569] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 1 is -OH. [0570] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein X 1 is C 2 -C 6 alkylenyl. [0571] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein X 2 is -CH 2 CH 2 -. [0572] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein X 4 is C 2 -C 6 alkylenyl. [0573] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein X 5 is C 2 -C 6 alkylenyl. [0574] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein Y 1 is: . [0575] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein Y 2 is: . [0576] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein each Z 3 is independently optionally substituted C1-C6 alkylenyl. [0577] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein each Z 3 is -CH2CH2-. [0578] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 6 is C5-C14 alkyl. [0579] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 7 is C5-C14 alkyl. [0580] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 6 is C6-C14 alkenyl. [0581] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 7 is C6-C14 alkenyl. [0582] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 8 is C5-C16 alkyl. [0583] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 9 is C5-C14 alkyl. [0584] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 8 is C 6 -C 14 alkenyl. [0585] In some embodiments, Lipids of the Disclosure have a structure of Formula (CY-VI’), or a pharmaceutically acceptable salt thereof, wherein R 9 is C 6 -C 14 alkenyl. [0586] In some embodiments, Lipids of the Disclosure comprise a heterocyclic core, wherein the heteroatom is nitrogen. In some embodiments, the heterocyclic core comprises pyrrolidine or a derivative thereof. In some embodiments, the heterocyclic core comprises piperidine or a derivative thereof. In some embodiments, Lipids of the Disclosure are selected from any lipid in Table (I) below or a pharmaceutically acceptable salt thereof:
Table (I). Non-Limiting Examples of Ionizable Lipids with a Cyclic Core
IV. DELIVERY VEHICLES AND TRACKING SYSTEMS [0587] Originator constructs and benchmark constructs described herein may be formulated in a delivery vehicle. Non-limiting examples of delivery vehicles include lipid nanoparticles, non- lipid nanoparticles, exosomes, liposomes, micelles, viral particles, and polymeric delivery technology. [0588] In some embodiments, the delivery vehicle comprises at least one lipid in Table (I). [0589] In some embodiments, the delivery vehicle comprises at least two lipids in Table (I). [0590] In some embodiments, the delivery vehicle comprises at least three lipids in Table (I). [0591] In some embodiments, the delivery vehicle comprises at least four lipids in Table (I). [0592] The total weight percentage of the lipid(s) in Table (I) in the delivery vehicle is between about 10% to about 95%, such as between about 10% to about 20%, between about 21% to about 30%, between about 31% to about 40%, between about 41% to about 50%, between about 51% to about 60%, between about 61% to about 70%, between about 71% to about 80%, between about 81% to about 90%, or between about 91% to about 95%. [0593] The total mole percentage of the lipid(s) in Table (I) in the delivery vehicle is between about 10% to about 95%, such as between about 10% to about 20%, between about 21% to about 30%, between about 31% to about 40%, between about 41% to about 50%, between about 51% to about 60%, between about 61% to about 70%, between about 71% to about 80%, between about 81% to about 90%, or between about 91% to about 95%. [0594] In some embodiments, at least one lipid in the delivery vehicle has a structure of Formula (CY-I), (CY-II), (CY-III), or (CY-IV). [0595] In some embodiments, at least two lipids in the delivery vehicle have a structure of Formula (CY-I), (CY-II), (CY-III), or (CY-IV). [0596] In some embodiments, at least three lipids in the delivery vehicle have a structure of Formula (CY-I), (CY-II), (CY-III), or (CY-IV). [0597] In some embodiments, at least four lipids in the delivery vehicle have a structure of Formula (CY-I), (CY-II), (CY-III), or (CY-IV). [0598] The total weight percentage of the lipid(s) having a structure of Formula (CY-I), (CY- II), (CY-III), or (CY-IV) in the delivery vehicle is between 10%-95%, such as between about 10% to about 20%, between about 21% to about 30%, between about 31% to about 40%, between about 41% to about 50%, between about 51% to about 60%, between about 61% to about 70%, between about 71% to about 80%, between about 81% to about 90%, or between about 91% to about 95%. [0599] The total mole percentage of the lipid(s) having a structure of Formula (CY-I), (CY-II), (CY-III), or (CY-IV) in the delivery vehicle is between 10%-95%, such as between about 10% to about 20%, between about 21% to about 30%, between about 31% to about 40%, between about 41% to about 50%, between about 51% to about 60%, between about 61% to about 70%, between about 71% to about 80%, between about 81% to about 90%, or between about 91% to about 95%. [0600] In some embodiments, the delivery vehicle further comprises at lease one additional lipid. Non-limiting examples include an additional cationic lipid, a neutral lipid, an anionic lipid, a helper lipid, a stealth lipid, or a polyethylene glycol (PEG) lipid. [0601] "Helper lipids" are lipids that enhance transfection, such as transfection of the delivery vehicle including the payloads and cargos. The mechanism by which the helper lipid enhances transfection may include enhancing particle stability and/or enhancing membrane fusogenicity. Helper lipids include steroids and alkyl resorcinols. Helper lipids suitable for use in the present disclosure include, but are not limited to, cholesterol, 5-heptadecylresorcinol, and cholesterol hemisuccinate. [0602] "Stealth lipids" are lipids that extend the length of time for which the delivery vehicle can exist in vivo (e.g. in the blood). Stealth lipids suitable for use in a lipid composition of the present disclosure include, but are not limited to, stealth lipids having a hydrophilic head group linked to a lipid moiety. [0603] Non-limiting examples of cationic lipids suitable for use in the delivery vehicle of the present disclosure include, but are not limited to, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3- dioleoyloxy) propyl)-N,N,N-trimethylammonium chloride (DOTAP), 1,2-Dioleoyl-3- Dimethylammonium-propane (DODAP), N-(1-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTMA), 1,2-Dioleoylcarbamyl-3-Dimethylammonium- propane (DOCDAP), 1,2-Dilineoyl-3-Dimethylammonium-propane (DLINDAP), dilauryl(C12:0) trimethyl ammonium propane (DLTAP), Dioctadecylamidoglycyl spermine (DOGS), DC-Choi, Dioleoyloxy-N-[2-sperminecarboxamido)ethyl}-N,N-dimethyl-1- propanaminiumtrifluoroacetate (DOSPA), 1,2-Dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide (DMRIE), 3-Dimethylamino-2-(Cholest-5-en-3-beta-oxybutan-4-oxy)-1- (cis,cis-9,12-octadecadienoxy)propane (CLinDMA), N,N-dimethyl-2,3- dioleyloxy)propylamine (DODMA), 2-[5′-(cholest-5-en-3[beta]-oxy)-3′-oxapentoxy)-3- dimethyl-1-(cis,cis-9′,12′-octadecadienoxy) propane (CpLinDMA) and N,N-Dimethyl-3,4- dioleyloxybenzylamine (DMOBA), and 1,2-N,N′-Dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP). [0604] Non-limiting example of neutral lipids suitable for use in the delivery vehicle of the present disclosure include a variety of neutral, uncharged or zwitterionic lipids. Examples of neutral phospholipids suitable for use in the present disclosure include, but are not limited to: 5-heptadecylbenzene-1,3-diol (resorcinol), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), phosphocholine (DOPC), dimyristoylphosphatidylcholine (DMPC), phosphatidylcholine (PLPC), 1,2-distearoyl-sn- glycero-3-phosphocholine (DAPC), phosphatidylethanolamine (PE), egg phosphatidylcholine (EPC), dilauryloylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), 1- myristoyl-2-palmitoyl phosphatidylcholine (MPPC), 1-palmitoyl-2-myristoyl phosphatidylcholine (PMPC), 1-palmitoyl-2-stearoyl phosphatidylcholine (PSPC), 1,2- diarachidoyl-sn-glycero-3-phosphocholine (DBPC), 1-stearoyl-2-palmitoyl phosphatidylcholine (SPPC), 1,2-dieicosenoyl-sn-glycero-3-phosphocholine (DEPC), palmitoyloleoyl phosphatidylcholine (POPC), lysophosphatidyl choline, dioleoyl phosphatidylethanolamine (DOPE), dilinoleoylphosphatidylcholine distearoylphophatidylethanolamine (DSPE), dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), palmitoyloleoyl phosphatidylethanolamine (POPE), lysophosphatidylethanolamine and combinations thereof. [0605] Non-limiting examples of anionic lipids suitable for use in the delivery vehicle of the present disclosure include, but are not limited to, phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoyl phosphatidyl ethanoloamine, N-succinyl phosphatidylethanolamine, N-glutaryl phosphatidylethanolamine cholesterol hemisuccinate (CHEMS), and lysylphosphatidylglycerol. [0606] In some embodiments, the weight ratio of the delivery vehicle (including all the lipids) and the payload is between about 100:1 to about 1:1, such as between about 100:1 to about 90:1, between about 89:1 to about 80:1, between about 79:1 to about 70:1, between about 69:1 to about 60:1, between about 59:1 to about 50:1, between about 49:1 to about 40:1, between about 39:1 to about 30:1, between about 29:1 to about 20:1, between about 19:1 to about 10:1, and between about 9:1 to about 1:1. [0607] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload. The cargo or payload may be any DNA, RNA or polypeptide described herein. [0608] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is a coding RNA. [0609] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is a non-coding RNA. [0610] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is a oRNA. [0611] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is an mRNA. [0612] In some embodiments, the at least one RNA compound is comprised of a functional RNA where the RNA results in at least one change in a cell, tissue, organ and/or organism. Said changes in state may include, but are not limited to, altering the expression level of a polypeptide, altering the translation level of a nucleic acid, altering the expression level of a nucleic acid, altering the amount of a polypeptide present in a cell, tissue, organ and/or organism, changing a genetic sequence of a cell, tissue, organ and/or organism, adding nucleic acids to a target genome, subtracting nucleic acids from a target genome, altering physiological activity in a cell, tissue, organ and/or organism or any combination thereof. [0613] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is DNA. [0614] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads which are DNA. The DNA may be the same DNA or different DNA. As a non-limiting example, the DNA are the same. As a non-limiting example, the DNA are different. As a non-limiting example, the DNA are different but encode the same payload or cargo. As a non-limiting example, the DNA are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0615] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with three cargos or payloads which are DNA. The DNA may be the same DNA or different DNA. As a non-limiting example, the DNA are the same. As a non-limiting example, the DNA are different. As a non-limiting example, two DNA are the same and one is different. As a non-limiting example, the first DNA is different from the second and third DNA. As a non-limiting example, the first DNA, second DNA and third DNA are all different. As a non-limiting example, the first DNA is different from the second and third DNA but they all encode the same payload or cargo. As a non-limiting example, the first DNA is different from the second and third DNA but the second and third DNA encode the same payload or cargo. [0616] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is a polypeptide. [0617] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads which are polypeptide. The polypeptide may be the same polypeptide or different polypeptide As a non-limiting example, the polypeptide are the same. As a non-limiting example, the polypeptide are different. As a non-limiting example, the polypeptides are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0618] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with three cargos or payloads which are polypeptide. The polypeptide may be the same polypeptide or different polypeptide. As a non-limiting example, the polypeptide are the same. As a non-limiting example, the polypeptide are different. As a non- limiting example, two polypeptide are the same and one is different. As a non-limiting example, the first polypeptide is different from the second and third polypeptide. As a non-limiting example, the first polypeptide, second polypeptide and third polypeptide are all different. As a non-limiting example, the first polypeptide is different from the second and third polypeptide but they all encode the same payload or cargo. As a non-limiting example, the first polypeptide is different from the second and third polypeptide but the second and third polypeptide encode the same payload or cargo. [0619] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is a peptide. [0620] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads which are peptide. The peptide may be the same peptide or different peptide. As a non-limiting example, the peptide are the same. As a non-limiting example, the peptides are different. As a non-limiting example, the peptides are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0621] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with three cargos or payloads which are peptide. The peptide may be the same peptide or different peptide. As a non-limiting example, the peptides are the same. As a non-limiting example, the peptides are different. As a non-limiting example, two peptides are the same and one is different. As a non-limiting example, the first peptide is different from the second and third peptide. As a non-limiting example, the first peptide, second peptide and third peptide are all different. As a non-limiting example, the first peptide is different from the second and third peptide but they all encode the same payload or cargo. As a non-limiting example, the first peptide is different from the second and third peptide but the second and third peptide encode the same payload or cargo. [0622] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with at least one cargo or payload which is RNA. [0623] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads which are RNA. The RNA may be the same RNA or different RNA. As a non-limiting example, the RNAs are the same. As a non-limiting example, the RNAs are different. As a non-limiting example, the RNAs are different but encode the same payload or cargo. As a non-limiting example, the RNAs are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0624] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with three cargos or payloads which are RNA. The RNA may be the same RNA or different RNA. As a non-limiting example, the RNA are the same. As a non-limiting example, the RNA are different. As a non-limiting example, two RNA are the same and one is different. As a non-limiting example, the first RNA is different from the second and third RNA. As a non-limiting example, the first RNA, second RNA and third RNA are all different. As a non-limiting example, the first RNA is different from the second and third RNA but they all encode the same payload or cargo. As a non-limiting example, the first RNA is different from the second and third RNA but the second and third RNA encode the same payload or cargo. [0625] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads where one is RNA and one is DNA. The RNA and DNA may encode the same peptide or polypeptide or may encode different peptides or polypeptides. As a non-limiting example, the RNA and DNA may encode the same peptide or polypeptide. As a non-limiting example, the RNA and DNA may encode different peptides or polypeptides. As a non-limiting example, the RNA and DNA are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0626] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads where one is RNA and one is a peptide. The RNA may encode the same peptide as the peptide cargo/payload the RNA may encode a different peptide. As a non-limiting example, the RNA encodes the same peptide. As a non- limiting example, the RNA encodes a different peptides. As a non-limiting example, the RNA and peptide are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0627] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads where one is RNA and one is a polypeptide. The RNA may encode the same polypeptide as the polypeptide cargo/payload the RNA may encode a different polypeptide. As a non-limiting example, the RNA encodes the same polypeptide. As a non-limiting example, the RNA encodes a different polypeptide. As a non- limiting example, the RNA and polypeptide are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0628] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads where one is DNA and one is a peptide. The DNA may encode the same peptide as the peptide cargo/payload the DNA may encode a different peptide. As a non-limiting example, the DNA encodes the same peptide. As a non- limiting example, the DNA encodes a different peptide. As a non-limiting example, the DNA and peptide are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). [0629] In some embodiments, the delivery vehicle comprises an originator construct or a benchmark construct with two cargos or payloads where one is DNA and one is a polypeptide. The DNA may encode the same polypeptide as the polypeptide cargo/payload the DNA may encode a different polypeptide. As a non-limiting example, the DNA encodes the same polypeptide. As a non-limiting example, the DNA encodes a different polypeptide. As a non- limiting example, the DNA and polypeptide are different pieces of a larger payload or cargo (e.g., heavy chain or light chain of an antibody) that can come together using natural systems or synthetic methods known in the art to produce a functional polypeptide (e.g., antibody). Delivery Vehicles Nanoparticles [0630] In some embodiments, the delivery vehicle is a nanoparticle. The term "nanoparticle" as used herein refers to any particle ranging in size from 10-1000 nm. The nanoparticle may be 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, or 1000 nm. Lipid Nanoparticles [0631] In some embodiments, the nanoparticles may be a lipid nanoparticle (LNP). In general, LNPs can be characterized as small solid or semi-solid particles possessing an exterior lipid layer with a hydrophilic exterior surface that is exposed to the non-LNP environment, an interior space which may aqueous (vesicle like) or non-aqueous (micelle like), and at least one hydrophobic inter-membrane space. LNP membranes may be lamellar or non-lamellar and may be comprised of 1, 2, 3, 4, 5 or more layers. In some embodiments, LNPs may comprise a cargo or a payload into their interior space, into the inter membrane space, onto their exterior surface, or any combination thereof. [0632] LNPs useful herein are known in the art and generally comprise cholesterol (aids in stability and promotes membrane fusion), a phospholipid (which provides structure to the LNP bilayer and also may aid in endosomal escape), a polyethylene glycol (PEG) derivative (which reduces LNP aggregation and "shields" the LNP from non-specific endocytosis by immune cells), and an ionizable lipid (complexes negatively charged RNA and enhances endosomal escape), which form the LNP-forming composition. [0633] The components of the LNP may be selected based on the desired target, cargo, size, etc. As a non-limiting example, previous studies have shown that that polymeric nanoparticles made of low molecular weight polyamines and lipids can deliver nucleic acids to endothelial cells with high efficiency. (Dahlman, et al., In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight, Nat Nanotechnol. 2014 Aug; 9(8): 648-655; the contents of which is herein incorporated by reference in its entirety). [0634] In some embodiments, the originator constructs and benchmark constructs of the present disclosure may be incorporated into lipid nanoparticles (LNPs). In some embodiments a lipid nanoparticle may be comprised of at least one cationic lipid, at least one non-cationic lipid, at least one sterol, at least one particle-activity-modifying-agent, or any combination thereof. In some embodiments a lipid nanoparticle may be comprised of at least one cationic lipid, at least one non-cationic lipid, at least one sterol, and at least one particle-activity- modifying-agent. In some embodiments, the LNP may be comprised of at least one cationic lipid, at least one non-cationic lipid, and at least one sterol. In some embodiments, the LNP may be comprised of at least one cationic lipid, at least one non-cationic lipid, and at least one particle-activity-modifying-agent. In some embodiments, the LNP may be comprised of at least one non-cationic lipid, at least one sterol, and at least one particle-activity-modifying-agent. In some embodiments, the LNP may be comprised of at least one cationic lipid and at least one non-cationic lipid. In some embodiments, the LNP may be comprised of at least one cationic lipid and at least one sterol. In some embodiments, the LNP may be comprised of at least one cationic lipid and at least one particle-activity-modifying-agent. In some embodiments, the LNP may be comprised of at least one non-cationic lipid and at least one sterol. In some embodiments, the LNP may be comprised of at least one non-cationic lipid and at least one particle-activity-modifying-agent. In some embodiments, the LNP may be comprised of at least one sterol and at least one particle-activity-modifying-agent. In some embodiments, the LNP may be comprised of at least one cationic lipid. In some embodiments, the LNP may be comprised of at least one non-cationic lipid. In some embodiments, a LNP may be comprised of a sterol. In some embodiments, the LNP may be comprised of a particle-activity-modifying- agent. [0635] In some embodiments, the at least one cationic lipid may comprise any of at least one ionizable cationic lipid, at least one amino lipid, at least one saturated cationic lipid, at least one unsaturated cationic lipid, at least one zwitterionic lipid, at least one multivalent cationic lipid, or any combination thereof. In some embodiments, the LNP may be essentially devoid of the at least one cationic lipid. In some embodiments, the LNP may contain no amount of the at least one cationic lipid. [0636] In some embodiments, at least one cationic lipid may be selected from, but not limited to, at least one of 1,3-Bis-(l,2-bis-tetradecyloxy-propyl-3- dimethylethoxyammoniumbromide)-propan-2-ol ((R)-PLC-2), 2-(Dinonylamino)ethan-1-ol (17-10), 2-(Didodecylamino)ethan-1-ol (17-11), 3-(Didodecylamino)propan-1-ol (17-12), 4- (Didodecylamino)butan-1-ol (17-13), 2-(Hexyl((9Z,12Z)-octadeca-9,12-dien-1- yl)amino)ethan-1-ol (17-2), 2-(Nonyl((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)ethan-1-ol (17-3), 2-(Dodecyl((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)ethan-1-ol (17-4), 2-(((9Z,12Z)- Octadeca-9,12-dien-1-yl)(tetradecyl)amino)ethan-1-ol (17-5), 2-(((9Z,12Z)-Octadeca-9,12- dien-1-yl)(octadecyl)amino)ethan-1-ol (17-6), 2-(Ditetradecylamino)ethan-1-ol (17-7), 2- (Di((Z)-octadec-9-en-1-yl)amino)ethan-1-ol (17-8), (9Z,12Z)-N-(2-Methoxyethyl)-N- ((9Z,12Z)-octadeca-9,12-dien-1-yl)octadeca-9,12-dien-1-amine (17-9), N-Nonyl-N-(2- (piperazin-1-yl)ethyl)nonan-1-amine (19-1), N-Dodecyl-N-(2-(piperazin-1-yl)ethyl)dodecan- 1-amine (19-2), (9Z,12Z)-N-((9Z,12Z)-Octadeca-9,12-dien-1-yl)-N-(2-(piperazi n-1- yl)ethyl)octadeca-9,12-dien-1-amine (19-3), N-Dodecyl-N-(2-(4-methylpiperazin-1- yl)ethyl)dodecan-1-amine1ntermediate1:2-(Didodecylamino)etha n-1-ol (19-4), N-Dodecyl- N-(2-(4-(4-methoxybenzyl)piperazin-1-yl)ethyl)dodecan-1-amin e (19-5), (9Z,12Z)-N-(2-(4- Dodecylpiperazin-1-yl)ethyl)-N-((9Z,12Z)-octadeca-9,12-dien- 1-yl)octadeca-9,12-dien-1- amine (19-6), (3-((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ylox y)-N,N- dimethylpropan-1-amine) (1-Bl 1), N-(2-(Didodecylamino)ethyl)-N-dodecylglycine (20-1), Dinonyl8,8'-((2-(dodecyl(2-hydroxyethyl)amino)ethyl)azanediy l)dioctanoate (20-10), 3-((2- (Ditetradecylamino)ethyl)(dodecyl)amino)propan-1-ol (20-11), 2-((2- (Ditetradecylamino)ethyl)(tetradecyl)amino)ethan-1-ol (20-12), 2-((2-(Di((9Z,12Z)-octadeca- 9,12-dien-1-yl)amino)ethyl)(dodecyl)amino)ethan-1-ol (20-13), 2-((2-(Di((9Z,12Z)-octadeca- 9,12-dien-1-yl)amino)ethyl)((9Z,12Z)-octadeca-9,12-dien-1-yl )amino)ethan-1-ol (20-14), 2- ((2-(Didodecylamino)ethyl)(hexyl)amino)ethan-1-ol (20-15), 2-((2- (Dinonylamino)ethyl)(nonyl)amino)ethan-1-ol (20-16), 2-((2- (Didodecylamino)ethyl)(nonyl)amino)ethan-1-ol (20-17), 2-((2- (Dinonylamino)ethyl)(dodecyl)amino)ethan-1-ol (20-18), 2-((2- (Didodecylamino)ethyl)amino)ethan-1-ol (20-19), Pentyl6-(dodecyl(2-(dodecyl(2- hydroxyethyl)amino)ethyl)amino)hexanoate (20-2), 2-((2- (Didodecylamino)ethyl)(dodecyl)amino)ethan-1-ol (20-20), 3-((2- (Didodecylamino)ethyl)(dodecyl)amino)propan-1-ol (20-21), 4-((2- (Didodecylamino)ethyl)(dodecyl)amino)butan-1-ol (20-22), (Z)-2-((2- (Didodecylamino)ethyl)(dodec-6-en-1-yl)amino)ethan-1-ol (20-23), 2-((2- (Didodecylamino)ethyl)(tetradecyl)amino)ethan-1-ol (20-24), 2-((2- (Didodecylamino)ethyl)((9Z,12Z)-octadeca-9,12-dien-1-yl)amin o)ethan-1-ol (20-25), Pentyl6-((2-(didodecylamino)ethyl)(2-hydroxyethyl)amino)hexa noate (20-3), Dipentyl6,6'- ((2-(dodecyl(2-hydroxyethyl)amino)ethyl)azanediyl)dihexanoat e (20-4), Diheptyl6,6'-((2-((6- (heptyloxy)-6-oxohexyl)(2hydroxyethyl)amino)ethyl)azanediyl) dihexanoate (20-5), Pentyl6- ((2-(dinonylamino)ethyl)(2-hydroxyethyl)amino)hexanoate (20-6), Heptyl6-(dodecyl(2- (dodecyl(2-hydroxyethyl)amino)ethyl)amino)hexanoate (20-7), Nonyl8-((2- (didodecylamino)ethyl)(2-hydroxyethyl)amino)octanoate (20-8), Heptadecan-9-yl8-((2- (didodecylamino)ethyl)(2-hydroxyethyl)amino)octanoate (20-9), 1-(2,2-Di((9Z,12Z)- octadeca-9,12-dien-1-yl)cyclopropyl)-N,N-dimethylmethanamine (21-1), 3,3-Di((9Z,12Z)- octadeca-9,12-dien-1-yl)cyclobutyl4-(dimethylamino)butanoate (21-2), 3,3-Di((9Z,12Z)- octadeca-9,12-dien-1-yl)cyclopentyl3-(dimethylamino)propanoa te (21-3), 3,3-Di((9Z,12Z)- octadeca-9,12-dien-1-yl)cyclopentyl4-(dimethylamino)butanoat e (21-4), 1-(2,3-Di((8Z,11Z)- heptadeca-8,11-dien-1-yl)cyclopropyl)-N,N-dimethylmethanamin e (21-6), Unknown (75- 016B), poly{4-((2-(dimethylamino)ethyl)thio)tetrahydro-2H-pyran-2-o ne}-r-poly{4- (octylthio)tetrahydro-2H-pyran-2-one} (A7), (3aR5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)- octadeca-9,12-dienyl)tetrahydro-3aH-cyclopentad1,3dioxol-5-a mine (ALN100), (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dien yl)tetrahydro- 3aHcyclopenta[d][l,3]dioxol-5-amine (ALN1001), ((3aR,5s,6aS)-N,N-dimethyl-2,2- di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d ][1,3]dioxol-5-amine)) (ALNY-100), dimyristoyltrimethylammoniumpropane (Amino Lipid 6), Benzamiπdiπ- dialkyl-carboxylicacid (BADACA), N,N-dihydroxyethylmethyl-N-2- (cholesteryloxycarbonylamino)ethylammoniumbromide (BHEM-Chol), N,N-bis-(2- hydroxyethyl)-N-methyl-N-(2-cholesteryloxycarbonylamino-ethy l)ammoniumbromide (BHEM-Chol1), 2-{4-[(3β)-cholest-5-en-3-yloxy]butoxy}-iV?N-dimethyl-3-[(9 Z,12Z)- octadeca-9!12-dien-l-yloxy]propan-1-amine (Butyl-CLinDMA), (2JR)-2-{4-[(3β)-cholest-5- en-3-yloxy]butoxy}-Λr^dimethyl-3-[(9Z,12Z)-octadeca-9!12-di en-l-yloxyjpropan-1-amine (Butyl-CLinDMA (2R)), (25)-2-{4-[(3β)-cholest-5-en-3-y1oxy]butoxy}-iVy/V-dimethyl -3- [(9Z,12Z)-octadeca-9,12-dien-l-yloxy]propan-1-amine (Butyl-CLinDMA (2S)), 1,1'-(2-(4-(2- ((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amin o)ethyl)piperazin-l- yl)ethylazanediyl)didodecan-2-ol (C 12-200), 1,1'-((2-(4-(2-((2-(bis(2- hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)pip erazin-1- yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200), Cholesteryl-succinyl Silane (C2), (9Z,9'Z,12Z,12'Z)-2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -1,3-diylbis(octadeca- 9,12-dienoate) (Cationic Lipid A2), 9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyloctadeca-9,1 2-dienoate (Cationic Lipid A3), l-(3-cholesteryl)-oxycarbonyl-aminomethylimidazole (CHIM), [(2-Morpholine-4-yl- ethylcarbamoyl)methyl]-carbamicacidcholesterylester (Chol-C3N-Mo2), [(2-Morpholine-4- yl-ethylcarbamoyl)-ethyl]-carbamicacidcholesterylesterChol-D MC3N-Mo2[l-Methyl-2-(2- morpholine-4-yl-ethylcarbamoyl)-propyl]-carbamicacidcholeste rylester (Chol-C4N-Mo2), l,17-bis(2-octylcyclopropyl)heptadecan-9-yl4-(dimethylamino) butanoate (CL), heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)-but anoate (CL01), cholesteryl3- (dimethylamino)propanoate (CL06), cholesteryl2-(dimethylamino)acetate (CL08), N,N- dimethyl-2,3-bis(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)propa n-1-amine (CL-1), N-methyl- 2-(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)-N-(2-((((9Z,12Z)-o ctadeca-9,12-diene-1- yl)oxy)ethyl)ethan-1-amine (CL-11), (3R,4R)-3,4-bis(((Z)-hexadec-9-en-1-yl)oxy)-1- methylpyrrolidine(CompoundCL-12) (CL-12), 2-(Dimethylamino)-N-((6Z,9Z,28Z,31Z)- Heptatriconta-6,9,28,31-tetraen-19-yl)acetamide (CL-13), 3-(Dimethylamino)propane-1,2- diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) (CL-14), (9Z,12Z)-di((9Z,12Z)-octadeca- 9,12-dien-1-yl)amine (CL-15), 7-Hydroxy7-(4-((1-methylpiperidine-4- carbonyl)oxy)butyl)tridecane-1,13-diyldidodecanoate (CL15B6), 7-Hydroxy7-(4-((1- methylpiperidine-4-carbonyl)oxy)butyl)tridecane-1,13-diyldit etradecanoate (CL15C6), 7- Hydroxy7-(4-((1-methylpiperidine-4-carbonyl)oxy)butyl)tridec ane-1,13-diyldipalmitate (CL15D6), 7-Hydroxy7-(4-((1-methylpiperidine-4-carbonyl)oxy)butyl)trid ecane-1,13- diyldioleate (CL15H6), Bis(2-(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)ethyl)amine (CL-16), (9Z,12Z)-N-Methyl-N-(2-(((9Z,12Z)-octadeca-9,12-dien-1-yl)ox y)ethyl)octadeca-9,12-dien- 1-amine (CL-17), (9Z,12Z)-N-(3-(((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)propyl) octadeca- 9,12-dien-1-amine (CL-18), (1-Methylpiperidin-3-yl)methyldi((11Z,14Z)-icosa-11,14-dien- 1- yl)carbamate (CL-19), N-methyl-N,N-bis(2-((Z)-hexadec-9-enyloxy)ethyl)amine (CL-2), (13Z,16Z)-N,N-Dimethyl-4-((9Z,12Z)-octadeca-9,12-dien-1-yl)d ocosa-3,13,16-trien-1-amine (CL-20), (S)-2-Amino-3-hydroxy-N,N-bis(2-(((Z)-octadeca-9-en-1- yl)oxy)ethyl)propanamide (CL-21), C2:N,N-dihexadecyl-N'-(3- triethoxysilylpropyl)succinamide (CL3), trans-1-Methyl-3,4-bis((((Z)-octadec-9-en-1- yl)oxy)methyl)pyrrolidine (CL-3), trans-1-methylpyrrolidine-3,4- diyl)bis(methylene)(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dieno ate) (CL-4), 7-(4- (Diisopropylamino)butyl)-7-hydroxytridecane-1,13-diylditetra decanoate (CL4C6), 7-(4- (Diisopropylamino)butyl)-7-hydroxytridecane-1,13-diyldipalmi tate (CL4D6), 11-(4- (Diisopropylamino)butyl)-11-hydroxyhenicosane-1,21-diyldiole ate (CL4H10), 7-(4- (Diisopropylamino)butyl)-7-hydroxytridecane-1,13-diyldioleat e (CL4H6), 9-(4- (Diisopropylamino)butyl)-7-hydroxyheptadecane-1,17-diyldiole ate (CL4H8), (6Z,9Z,28Z,31Z)-Heptatriaconta-6,9,28,31-tetraen-19-yl4-(dim ethylamino)butanoate (CL-5), 2-(Dimethylamino)-N-(2-(((Z)-octadeca-9-en-1-yl)oxy)ethyl)-N -((9Z,12Z)-octadeca-9,12- diene-1-yl)acetamide (CL-53), 3-((2-(((Z)-octadeca-9-en-1-yl)oxy)ethyl)((9Z,12Z)-octadeca- 9,12-dien-1-yl)amino)propane-1-All (CL-54), 1-Methyl-3,3-bis((((9Z,12Z)-octadeca-9,12- dien-1-yl)oxy)methyl)azetidine (CL-55), 1-Methyl-3,3-bis(2-(((9Z,12Z)-octadeca-9,12-dien- 1-yl)oxy)ethyl)azetidine (CL-56), 1-Methyl-3,3-bis(2-(((9Z,12Z)-octadeca-9,12-dien-1- yl)oxy)propyl)azetidine (CL-57), 2-(3,3-di((9Z,12Z)-octadeca-9,12-dien-1-yl)azetidin-1- yl)ethan-1-ol (CL-58), 2-(3,3-di((9Z,12Z)-octadeca-9,12-dien-1-yl)azetidin-1-yl)pro pan-1-ol (CL-59), 3-(Di((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)propan-1-o (CL-6), 3- (Dimethylamino)propyl3,3-di((9Z,12Z)-octadeca-9,12-dien-1-yl )azetidine-1-carboxylate (CL-60), 2-(Di((Z)-octadeca-9-en-1-yl)amino)ethane-1-ol (CL-61), 3-(Di((Z)-octadeca-9-en- 1-yl)amino)propan-1-ol (CL-62), (11Z,14Z)-2-((Dimethylamino)methyl)-2-((9Z,12Z)- octadeca-9,12-dien-1-yl)icosa-11,14-dien-1-ol (CL-63), (11Z,14Z)-2-(Dimethylamino)-2- ((9Z,12Z)-octadeca-9,12-dien-1-yl)icosa-11,14-dien-1-ol (CL-64), 3-(Dimethylamino)-2,2- bis((((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)methyl)propan-1-o l (CL-65), (9Z,12Z)-N-(2- (((Z)-Octadeca-9-en-1-yl)oxy)ethyl)octadeca-9,12-dien-1-amin e (CL-7), 1-Methyl-3,3- di((9Z,12Z)-octadeca-9,12-dien-1-yl)azetidine (CL-8), N,2-Dimethyl-1,3-bis(((9Z,12Z)- octadeca-9,12-dien-1-yl)oxy)propan-2-amine (CL-9), 3-Dimethylamino-2-(Cholest-5-en-3B- oxybutan-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (CLinDMA), 2-[5′-(cholest-5-en- 3-oxy)-3′-oxapentoxy)-3-dimethy-1-(cis,cis-9′,12′-octa decadienoxy)propane (CpLinDMA), cetyltrimethylammoniumbromide (CTAB), l^-Diarachidonyloxy-^TV-dimethy^-propyl-S- amine (DAraDMA), 0,0'-ditetradecanoyl-N-(α- trimethylammonioacetyl)diethanolaminechloride (DC-6-14), 3β-[N-(N′,N′- dimethylaminoethane)carbamoyl]cholesterol (DC-Chol), dimethyldioctadecylammonium (DDA), dimethyldioctadecylammoniumbromide (DDA ), N,N-distearyl-N,N- dimethylammoniumbromide (DDAB), l,2-Didocosahexaenyloxy-(7V,N-dimethyl)-propyl-3- amine (DDocDMA), N-(2-(dimethylamino)ethyl)-4,5-bis(dodecylthio)pentanamide (DEDPA), 3-Dimethylamino-2-(Cholest-5-en-3β-oxypent-3-oxa-an-5-oxy)- 1-(cis,cis-9,12- octadecadienoxy)propane (DEG-CLinDMA), 1,6-DileoylTriethylenetetramide (dio-TETA), Nl,N19-bis((S,23E,25E,27E,29E)-16-((2E,4E,6E,8E)-3,7-dimethy l-9-(2,6,6-trimethylcyclo- hex-l-en-l-yl)nona-2,4,6,8-tetraenamido)-24,28-dimethyl-15,2 2-dioxo-30-(2,6,6- trimethylcyclohex-l-en-l-yl)-4,7,10-trioxa-14,21-diazatriaco nta-23,25,27,29-tetraen-l-yl)- 4,7,10,13,16-pentaoxanonadecane-1,19-diamide (diVA-PEG-diVA), DiLin-N- Methylpiperazine (DL-033), DiLin-N,N-DimethylGlycine (DL-036), Dioleyl-N,N- DimethylGlycine (DL-048), 3-((1,3-bis(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propan-2- yl)amino)propanoicacid (DLAPA), 1,2-dilinolenyloxy-3-dimethylaminopropane (DLenDMA), 1-Linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 3-(N,N- Dilinoleylamino)-1,2-propanediol (DLinAP), 1,2-N,N′-Dilinoleylcarbamyl-3- dimethylaminopropane (DLincarbDAP), 1,2-Dilinoleoylcarbamyl-3-dimethylaminopropane (DLinCDAP), 1,2-Dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2- Dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-Dilinoleoyl-3- dimethylaminopropane (DLinDAP), 1,2-DiLinoleyloxy-N,N-dimethylaminopropane (DLinDMA ), 1,2-dilinoleyloxy-3-dimethylaminopropane (DLinDMA 1), 1,2-Dilinoleyloxo- 3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), dilinoleoyl-4-aminobutyricacid (DLinFAB), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-K-C2-DMA), 2,2-Dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), 1,2-Dilinoleyoxy-3- morpholinopropane (DLin-MA), (6Z,9Z,28Z,31Z)-heptatriacont-6,9,28,31-tetraene-19-yl4- (dimethylamino)butanoate (DLin-MC3-DMA), l,2-Dilinoleyloxy-3-(N- methylpiperazino)propane (DLinMPZ), 1,2-Dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), Dilinoleyloxy3-piperidinopropylamine (DLinPip), 1.2Dilinoleyloxy3-(3'- hvdroxypiperidino)-propylamine (DLinPip-3OH), 1,2Dilinoleyloxy3-(4'-hvdroxypiperidino)- propylamine (DLinPip-4OH), 1,2-Dilinoleyloxy-3-hvdroxypropane (DLinPO), 1,2- Dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), l,2-Dilinoleoyl-3- trimethylaminopropane (DLinTAP), 1,2-Dilinoleoyl-3-trimethylaminopropanechloridesalt (DLin-TAP.Cl), l,2-Dilinoleyloxy-3-trimethylaminopropane (DLinTMA), 1,2-Dilinoleyloxy- 3-trimethylaminopropanechloridesalt (DLin-TMA.Cl), 3-((1,3-bis(((9Z,12Z.15Z)-octadeca- 9,12,15-trienoyl)oxy)propan-2-yl)amino)propanoicacid (DLLAPA), 1,2Dilinoleyloxy3- (N,NdimethyD-propylamme (DLmDEA), l,2-Dilauroyl-sn-Glicero-3-Phosphoethanolamine (DLPE), l,2-Dilauroyl-sn-Glicero-3-Glycerol (DLPG), N,N-Dimethyl-3,4- dioleyloxybenzylamine (DMOBA), dimyristoylphosphatidylserine (DMPS), N-[l-(2,3- dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl)ammoniu mbromide (DMRIE), 1,2- Dimyristyloxypropyl-3-dimethyl-hydroxyethylammoniumbromide (DMRIE1), l,2- dimyristoyl-3-trimethylammoniumpropane (DMTAP), 3-(N,N-Dioleylamino)-1,2-propanedio (DOAP), 3-((1,3-bis(oleoyloxy)propan-2-yl)amino)propanoicacid (DOAPA), 1,2-N,N′- dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 1,2-Dioleoylcarbamyl-3- Dimethylammonium-propane (DOCDAP), N,N-dioleyl-N,N-dimethylammoniumchloride (DODAC), 1,2-Dioleoyl-3-Dimethylammonium-propane (DODAP), N,N- dihydroxyethylΝ,Ν-dioctadecylammoniumchloride (DODEAC), N,N-dimethyl-2,3- dioleyloxypropylamine (DODMA), dioleoyl-4-aminobutyricacid (DOFAB), Dioctadecylamidoglycylspermine (DOGS), 1,2-Dioleoyl-3-methyl-(methoxycarbonyl- ethyl)ammonium-Propane (DOMCAP), 1,2-Dioleoyl-3-N-pyrrolidine-propane (DOP5P), 1,2- Dioleoyl-3-N-pyrridinium-propane,bromidesalt (DOP6P), 1,2-dioleoyl-3-dimethyl- hydroxyethylammoniumbromide (DORI), 1,2-dioleyloxypropyl-3-dimethyl- hydroxyethylammoniumbromide (DORIE), 1,2-dioleyloxypropyl-3-dimethyl- hydroxybutylammoniumbromide (DORIE-HB), 1,2-dioleyloxypropyl-3-dimetyl- hydroxypropylammoniumbromide (DORIE-HP), 1,2-dioleyloxypropyl-3-dimethyl- hydroxypentylammoniumbromide (DORIE-Hpe), 2,3-dioleyloxy-N-[2(spermine- carboxamido)ethyl]-N,N-dimethyl-1-propanaminiumtrifluoroacet ate (DOSPA), 1,3- dioleoyloxy-2-(6-carboxy-spermyl)-propylamide (DOSPER), N-(1-(2,3-dioleoyloxy)propyl)- N,N,N-trimethylammoniumchloride (DOTAP), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP1), N-[5'-(2',3'-dioleoyl)uridine]-Ν',Ν',Ν'-trimethylammonium tosylate (DOTAU), 1- [2-(9(Z)-octadecenoyloxy)ethyl]-2-(8(Z)-heptadecenyl-3-(2- hydroxyethyl)imidazoliniumchloride (DOTIM), N-(1-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammoniumchloride (DOTMA), dioleylphosphatidyluridinephosphatidylcholine (DOUPC), 1,2-Diphvtanyloxy-W.N-dimemyl)-butyl-4-amme (DPan-C2-DMA), l,2- Diphytanyloxy-3-(iV,7V-dimethy1)-propylamine (DPanDMA), 2,3-bis(dodecylthio)propyl(2- (dimethylamino)ethyl)carbamate (DPDEC), dipalmitoyl-4-aminobutyricacid (DPFAB), 1,2- dipalmityloxypropyl-3-dimethyl-hydroxyethylammoniumbromide (DPRIE), 1,2-dipalmitoyl- 3-trimethylammoniumpropane (DPTAP), 1-[2-(hexadecanoyloxy)ethyl]-2-pentadecyl-3-(2- hydroxyethyl)imidazoliniumchloride (DPTIM), 3-((1,3-bis(stearoyloxy)propan-2- yl)amino)propanoicacid (DSAPA), distearyldimethylammonium (DSDMA), 1,2-distearloxy- N,N-dimethylaminopropane (DSDMA1), 1,2-disteryloxypropyl-3-dimethyl- hydroxyethylammoniumbromide (DSRIE), l,2-disteroyl-3-trimethylammoniumpropane (DSTAP), ditetradecyltrimethylammonium (DTDTMA), 1,2-dioleoyl-sn-glycero-3- ethylphosphocholine (EDOPC), N2-[N2,N5-bis(3-aminopropyl)-L-ormithyl]-N,N- dioctadecyl-L-glutaminetetrahydrotrifluoroacetate (GC33), Cholest-5-en-3-ol(3P)-,3-[(3- aminopropyl)[4-[(3-aminopropyl)amino]butyl]carbamate] (GL67), glycerylmono-oleate (GMO), Guanidino-dialkyl-carboxylicacid (GUADACA), 2-(bis(2- (tetradecanoyloxy)ethyl)amino)-N-(2-hydroxyethyl)-N,N-dimeth yl-2-oxoethan- aminiumbromide (HEDC), 2,2'-(tert-butoxycarbonylazanediyl)bis(ethane-2,1- diyl)ditetradecanoate (HEDC-BOC-TN), 1-(2-(((3S,10R,13R)-10,13-dimethyl-17-((R)-6- methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetra decahydro-1H- cyclopenta[a]phenanthren-3-yldisulfanyl)ethyl)guanidine (HGT4002), (15Z,18Z)-N,N- dimethyl-6-(9Z,12Z)-octadeca-9,12-dien-l-yl)tetracosa-l5,18- dien-l-amine (HGT5000), (15Z,18Z)-N,N-dimethyl-6-((9Z,12Z)-octadeca-9,12-dien-l-yl)t etracosa-4,15,18-trien-l- amine (HGT5001), Histaminyl-Cholesterolhemisuccinat (HisChol), histidinylcholesterolhemisuccinate (Hist-Chol), HydroSoyPC (HSPC), imidazolecholesterolester (ICE), 3-(didodecylamino)-N1,N1,4-tridodecyl-1- piperazineethanamine (KL10), N1-[2-(didodecylamino)ethyl]-N1,N4,N4-tridodecyl-1,4- piperazinediethanamine (KL22), 14,25-ditridecyl-15,18,21,24-tetraaza-octatriacontane (KL25), N,N-di-n-lctradecyl,N-methyl-N-(2-guanidinyl)cthylammonium (Lipid 1), N,N-di-n- octadecyl,N-mcthyl-N-(2-guanidinyl)cthylammoniumchloride (Lipid 2), 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-oct adeca-9,12-dienoate (Lipid A), (9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyloctadeca-9,1 2-dienoate (Lipid A1), 2,2- Dilinoleyl-4-dimethylaminoethyl-[l,3]-dioxolane (Lipid A2), ((5-((dimethylamino)methyl)- l,3-phenylene)bis(oxy))bis(octane-8,l-diyl)bis(decanoate) (Lipid B), 2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -l,3-diyl9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) (Lipid C), 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)-13- (octanoyloxy)tridecyl3-octylundecanoate (Lipid D), (6Z,16Z)-12-((Z)-dec-4-en-1-yl)docosa- 6,16-dien-11-yl5-(dimethylamino)pentanoate (Lipid I), Dioctadecyl-(2-hydroxy1-3- propylamino)aminopolylysine (Lipid T), (3-((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31- tetraen-19-yloxy)-N,N-dimethylpropan-1-amine (MC3 Ether), describedinU.S.ProvisionalApplicationNo.61/384,050 (MC3 Thioester), (4- ((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yloxy)- N,N-dimethylbutan-1-amine (MC4 Ether), 3-((2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)ethyl)amino)propa noicacid (MLAPA), 3-((2-(((9Z,12Z,15Z)-octadeca-9,12,15-trienoyl)oxy)ethvnamin o)propanoicacid (MLLAPA), mon-omycolylglycerol (MMG), 3-((2-(oleoyloxy)ethyl)amino)propanoicacid (MOAPA), 4-(2-Aminoethyl)-Morpholino-Cholesterolhemisuccinat (MoChol), 1,2-Dioleoyl- 3-N-morpholine-propane (MoDO), Methylpyridiyl-dialkyl-carboxylicacid (MPDACA), monopalmitoylphosphatidylcholin (MPPC), 3-((2-(stearoyloxy)ethyl)amino)propanoicacid (MSAPA), N1-[2-((lS)-1-[(3-aminopropyl)amino]-4-[di(3-amino- propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzam ide (MVL5), 2-({8-[(3β)- cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-oct adeca-9,12-dien-1- yloxy]propan-1-amine (Octyl-CLinDMA), (2R)-2-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)- N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1 -amine (Octyl-CLinDMA (2R)), phosphatidylcholines (PC), l,3-Bis-(l,2-bis-tetradecyloxy-propyl-3- dimemylethoxyammoniumbromide)-propane-2-ol (PCL-2), palmitoyi-oieoyl-nor-arginine (PONA), stearylamine (STA), 2-(((tert-Butyldimethylsilyl)oxy)methyl)-2- (hydroxymethyl)propane-1,3-diol (Synthesis Example 1 (A)), 3-((tert- Butyl(dimethyl)silyl)oxy)-2,2-bis(((9Z)-tetradec-9-enoyloxy) methyl)propyl(9Z)-tetradec-9- enoate (Synthesis Example 1 (B)), 3-Hydroxy-2,2-bis(((9Z)-tetradec-9- enoyloxy)methyl)propyl(9Z)-tetradec-9-enoate (Synthesis Example 1 (C)), 3-((4- (Dimethylamino)butanoyl)oxy)-2,2-bis(((9Z)-tetradec-9-enoylo xy)methyl)propyl(9Z)- tetradec-9-enoate (Synthesis Example 1 (D)), 3-(5-(bis(2-hydroxydodecyl)amino)pentan-2- yl)-6-(5-((2-hydroxydodecyl)(2-hydroxyundecyl)amino)pentan-2 -yl)-l,4-dioxane-2,5-dione) (Target 24), trehalose6'6'-dibehenate (TDB), 1,1'-(2-(4-(2-((2- (bis(2hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)eth yl)piperazin-1- yl)ethylazanediyl)didodecan-2-ol (Tech G1), 3-((1,3-bis(((9Z,12Z)-octadeca-9,12- dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl) propan-2- yl)amino)propanoicacid (TLAPA), (l-(2,3-linoleyloxypropoxy)-2-(linoleyloxy)-(7V,Λ/- dimethyl)-propyl-3-amine) (TLinDMA), 3-((1,3-bis(((9Z.12Z.15Z)-octadeca-9.12.15- trienoyl)oxy)-2-((((9Z.12Z.15E)-octadeca-9,12,15-trienoyl)ox y)methyl)propan-2- yl)amino)propanoicacid (TLLAPA), N-(α-trimethylammonioacetyl)-didodecyl-D- glutamatechloride (TMAG), 3-((1,3-bis(((Z)-octadec-9-enoyl)oxy)-2-((((Z)-octadec-9- enoyl)oxy)methyl)propan-2-yl)amino)propanoicacid (TOAPA), 3-((1,3-bis(stearoyloxy)-2- ((stearoyloxy)methyl)propan-2-yl)amino)propanoicacid (TSAPA), 1,N19- bis((16E,18E,20E,22E)-17,21-dimethyl-15-oxo-23-(2,6,6-trimet hylcyclohex-1-en-1-yl)- 4,7,10-trioxa-14-azatricosa-16,18,20,22-tetraen-1-yl)-4,7,10 ,13,16-pentaoxanonadecane- 1,19-diamide (VA-PEG-VA), 2,2-Dillinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (XTC), disclosedinNon-PatentLiterature11 (YSK05), 1,2-di-γ-linolenyloxy-N,N- dimethylaminopropane (γ-DLenDMA), a-D-Tocopherolhemisuccinoyl, (9Z,9,Z,12Z,12,Z)-2- ((2-(((3-(dimethylamino)propoxy)carbonyl)oxy)tetradecanoyl)o xy)propane-1,3- diylbis(octadeca-9,12-dienoate), 2-(((13Z,16Z)-4-(((3- (diethylamino)propoxy)carbonyl)oxy)docosa-13,16-dienoyl)oxy) propane-1,3- diyldioctanoate, 2-(((13Z,16Z)-4-(((3-(dimethylamino)propoxy)carbonyl)oxy)doc osa-13,16- dienoyl)oxy)propane-1,3-diyldioctanoate, 2-((4-(((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)pr opane-1,3-diyldioctanoate, 2-((4-(((3-(ethyl(methyl)amino)propoxy)carbonyl)oxy)hexadeca noyl)oxy)propane-1,3- diylbis(decanoate), 2-((4-(((3- (diethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane- 1,3-diylbis(decanoate), 2- (10-dodecyl-3-ethyl-8,14-dioxo-7,9,13-trioxa-3-azaicosan-20- yl)propane-1,3-diyldioctanoate, 2-(((4-(dimethylamino)butanoyl)oxy)methyl)-2-((octanoyloxy)m ethyl)propane-1,3- diyl(9Z,9′Z)bis-tetradec-9-enoate, (9Z,9'Z,12Z,12'Z)-2-(((1-(cyclopropylmethyl)piperidine-4- carbonyl)oxy)methyl)propane-1,3-diylbis(octadeca-9,12-dienoa te), ((2-(((1- isopropylpiperidine-4-carbonyl)oxy)methyl)-1,4-phenylene)bis (oxy))bis(octane-8,1- diyl)bis(decanoate), 2-((4-(((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)pr opane-1,3- diyldidodecanoate, 2-((4-(((3- (diethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane- 1,3-diyldidodecanoate, 2- ((4-(((3-(dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)ox y)propane-1,3- diyldidodecanoate, 2-((4-(((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)pr opane-1,3- diylditetradecanoate, 2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -1,3-diylditetradecanoate, 2-((4-(((3-(diethylamino)propoxy)carbonyl)oxy)hexadecanoyl)o xy)propane-1,3- diylditetradecanoate, (Z)-2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -1,3-diyldioleate, (9Z,9,Z,12Z,12,Z,15Z,15,Z)-2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -1,3-diylbis(octadeca- 9,12,15-trienoate), (9Z,9,Z,12Z,12,Z)-2-((4-(((3- (diethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane- 1,3-diylbis(octadeca-9,12- dienoate), (9Z,9,Z,12Z,12,Z)-2-((4-(((3- (dimethylamino)propoxy)carbonyl)oxy)hexadecanoyl)oxy)propane -1,3-diylbis(octadeca- 9,12-dienoate), N,N,N-trimethyl-5-oxo-5-(3-((3-pentyloctanoyl)oxy)-2,2-bis(( (3- pentyloctanoyl)oxy)methyl)propoxy)pentane-1-Aminiumiodide3-( (5- (dimethylamino)pentanoyl)oxy)-2,2-bis(((3-pentyloctanoyl)oxy )methyl)propyl3- pentyloctanoate, 3-dimethylaminopropylcarbonate(9Z,12Z)-octacosa-19,22-dien-1 1-yl, 2- (((N,N-dimethyl-β-alanyl)oxy]methyl}-2-[(octanoyloxy)methyl )propane-1,3-diyl(9Z,9′Z)bis- tetradec-9-enoate, ΟΊ,O1-(2-(7-dodecyl-14-methyl-3,9-dioxo-2,4,8,10-tetraoxa- 14- azapentadecyl)propane-1,3-diyl)8-dimethyldioctanedioate, 8-dimethylΟΊ,01-(2-(((1- methylpyrrolidine-3-carbonyl)oxy)methyl)propane-1,3-diyl)dio ctanedioate, 1-(3-((6,6-bis((2- propylpentyl)oxy)hexanoyl)oxy)-2-(((1,4-dimethylpiperidine-4 - carbonyl)oxy)methyl)propyl)8-methyloctanedioate, (9Z,12Z)-5-(((3- (dimethylamino)propoxy)carbonyl)oxy)-7-octylpentadecyloctade ca-9,12-dienoate, 5-(((3- (dimethylamino)propoxy)carbonyl)oxy)-7-octylpentadecyloctano ate, 1-(3-((6,6-bis((2- propylpentyl)oxy)hexanoyl)oxy)-2-(((1,4-dimethylpiperidine-4 - carbonyl)oxy)methyl)propyl)10-octyldecanedioate, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)-5-octyltridecyldecanoat e, 1-(16-(((4,4- bis(octyloxy)butanoyl)oxy)methyl)-9-dodecyl-2-methyl-7,13-di oxo-6,8,12,14-tetraoxa-2- azaheptadecan-17-yl)8-methyloctanedioate, 3-((5-(dimethylamino)pentanoyl)oxy)-2,2- bis(((9Z)-tetradec-9-enoyloxy)methyl)propyl(9Z,12Z)-octadec- 9,12-dienoate, 3-((5- (Dimethylamino)pentanoyl)oxy)-2,2-bis(((3-pentyloctanoyl)oxy )methyl)propyl3- pentyloctanoate, (9Z,9'Z,12Z,12'Z)-2-(((3-(diethylamino)propanoyl)oxy)methyl) propane-1,3- diylbis(octadeca-9,12-dienoate), ((2-(((4-(dimethylamino)butanoyl)oxy)methyl)-1,4- phenylene)bis(oxy))bis(octane-8,1-diyl)bis(decanoate), 1-(3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((1-methylpyrrolidine-3-carbon yl)oxy)methyl)propyl)8- methyloctanedioate, 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((palmitoyloxy)methyl)p ropyl1- methylpyrrolidine-3-carboxylate, 3-((4,4-bis(octyloxy)butanoyl)oxy)-2- ((tetradecanoyloxy)methyl)propyl1-methylpyrrolidine-3-carbox ylate, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)-13-(octanoyloxy)tridecy l9-pentyltetradecanoate, 3- ((4,4-bis(octyloxy)butanoyl)oxy)-2-((dodecanoyloxy)methyl)pr opyl1-methylpyrrolidine-3- carboxylate, 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)-13-hydroxytridec yl9- pentyltetradecanoate, 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)-13- (octanoyloxy)tridecyl7-hexyltridecanoate, 2-(5-(3-((1-methylpyrrolidine-3-carbonyl)oxy)-2- ((tetradecanoyloxy)methyl)propoxy)-5-oxopentyl)propane-1,3-d iyldioctanoate, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)-13-(octanoyloxy)tridecy l5-heptyldodecanoate, 2-(5- (3-((1-methylpyrrolidine-3-carbonyl)oxy)-2-((palmitoyloxy)me thyl)propoxy)-5- oxopentyl)propane-1,3-diyldioctanoate, 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)-13- hydroxytridecyl5-heptyldodecanoate, 2-(((1-methylpyrrolidine-3- carbonyl)oxy)methyl)propane-1,3-diylbis(6,6-bis(octyloxy)hex anoate), (9Z,12Z)-3-(((3- dimethylamino)propoxy)carbonyl)oxy)-13-(octanoyloxy)tridecyl octadeca-9,12-dienoate, 3- ((5-(dimethylamino)pentanoyl)oxy)-2,2-bis(((9Z)-tetradec-9-e noyloxy)methyl)propyl(9Z)- octadec-9-enoate, 2-(10-dodecyl-3-ethyl-8,14-dioxo-7,9,13-trioxa-3-azanonadeca n-19- yl)propane-1,3-diyldioctanoate, ((2-(((1-methylpiperidine-4-carbonyl)oxy)methyl)-1,4- phenylene)bis(oxy))bis(octane-8,1-diyl)bis(decanoate), 2-(((3- (dimethylamino)propanoyl)oxy)methyl)propane-1,3-diylbis(4,4- bis(octyloxy)butanoate), (9Z,12Z)-2-(((11Z,14Z)-2-((3-(dimethylamino)propanoyl)oxy)ic osa-11,14-dien-1- yl)oxy)ethyloctadeca-9,12-dienoate, 2-(((1,3-dimethylpyrrolidine-3- carbonyl)oxy)methyl)propane-1,3-diylbis(4,4-bis(octyloxy)but anoate), (13Z,16Z)-4-(((3- (dimethylamino)propoxy)carbonyl)oxy)docosa-13,16-dien-1-ylhe ptadecan-9-ylsuccinate, 2,2- bis(heptyloxy)ethyl3-((3-ethyl-10-((9Z,12Z)-octadeca-9,12-di en-1-yl)-8,15-dioxo-7,9,14- trioxa-3-azaheptadecan-17-yl)disulfanyl)propanoate, 2-(((1-methylpyrrolidine-3- carbonyl)oxy)methyl)propane-1,3-diylbis(4,4-bis(octyloxy)but a, 1-(3-((1,3- dimethylpyrrolidine-3-carbonyl)oxy)-2-(((9Z,12Z)-octadeca-9, 12- dienoyloxy)methyl)propyl)10-octyldecanedioate, (13Z,16Z)-4-(((3- (diethylamino)propoxy)carbonyl)oxy)docosa-13,16-dien-1-yl2,2 -bis(heptyloxy)acetate, (13Z,16Z)-4-(((2-(dimethylamino)ethoxy)carbonyl)oxy)docosa-1 3,16-dien-1-yl2,2- bis(heptyloxy)acetate, Aceticacid(20,23R)-2-methyl-9-[(9Z,12Z)-octadeca-9,12-dien-1 -yl]-7- oxo-6,8,11-trioxa-2-azanonacosa-20-En-23-yl3-(dimethylamino) propylcarbonate(11Z,14Z)- 1-{[(9Z,12R)-12-hydroxyoctadec-9-en-1-yl], (12Z,15Z)-1-((((9Z,12Z)-octadeca-9,12-dien-1- yloxy)carbonyl)oxy)henicosa-12,15-dien-3-yl3-(dimethylamino) propanoate, (9Z,12Z)-3- ((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (dimethylamino)propyl)carbamoyl)oxy)methyl)propyloctadeca-9, 12-dienoate, (12Z,15Z)-3- ((4-(dimethylamino)butanoyl)oxy)henicosa-12,15-dien-1-yl9-pe ntyltetradecanoate, (9Z,12Z)- 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((((1,2,2,6,6-pentamet hylpiperidin-4- yl)oxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (12Z,15Z)-3-((4- (dimethylamino)butanoyl)oxy)henicosa-12,15-dien-1-yl7-hexylt ridecanoate, (9Z,12Z)-3- ((4,4-bis(octyloxy)butanoyl)oxy)-2-(((((1-methylpiperidin-4- yl)methoxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (12Z,15Z)-3-((4- (dimethylamino)butanoyl)oxy)henicosa-12,15-dien-1-yl5-heptyl dodecanoate, (9Z,12Z)-3- ((4,4-bis(octyloxy)butanoyl)oxy)-2-(((((1-ethylpiperidin-4- yl)oxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (12Z,15Z)-3-((4- (dimethylamino)butanoyl)oxy)henicosa-12,15-dien-1-yl3-octylu ndecanoate,formatesalt, 3- ((5-(dimethylamino)pentanoyl)oxy)-2,2-bis(((9Z)-tetradec-9-e noyloxy)methyl)propyl(9Z)- hexadec-9-enoate, (9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((((1-methyla zetidin-3- yl)oxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (9Z,12Z)-(12Z,15Z)-3-((3- (dimethylamino)propanoyl)oxy)henicosa-12,15-dien-1-yloctadec a-9,12-dienoate, 2-(((3- (diethylamino)propoxy)carbonyl)oxy)tetradecyl4,4-bis((2-ethy lhexyl)oxy)butanoate, (9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((((1-methylp iperidin-4- yl)oxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((((1-methylpyrrolidin-3- yl)oxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (9Z,12Z)-3-(((2- (dimethylamino)ethoxy)carbonyl)oxy)pentadecyloctadeca-9,12-d ienoate, (9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(4-methylpiperazin-1- yl)propoxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, 3- (Dimethylamino)propyltriacontan-11-ylcarbonateTriacontan-11- ol, (9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(pyrrolidin-1- yl)propoxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, (9Z,12Z)-3-(((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)pentadecyloctadeca- 9,12-dienoate, 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl4- ((diethylamino)methyl)benzoate, (9Z,12Z)-3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyloctadeca-9,12-d ienoate, 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl3- ((dimethylamino)methyl)benzoate, (9Z,12Z)-3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyloctadeca-9,12- dienoate, 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl1- methylpiperidine-3-carboxylate, 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca- 9,12-dienoyloxy)methyl)propyl1-methylpiperidine-4-carboxylat e, 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl1,4- dimethylpiperidine-4-carboxylate, 3-((4-(dimethylamino)butanoyl)oxy)-2,2-bis(((9Z)- tetradec-9-enoyloxy)methyl)propyl(9Z)-hexadec-9-enoate, 2-(10-dodecyl-3-ethyl-8,14-dioxo- 7,9,13-trioxa-3-azahexadecan-16-yl)propane-1,3-diyldioctanoa te, (9Z,9'Z,12Z,12'Z)-2-(((4- (piperidin-1-yl)butanoyl)oxy)methyl)propane-1,3-diylbis(octa deca-9,12-dienoate), 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl4- methylmorpholine-2-carboxylate, (2R)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)- octadeca-9,12-dienoyloxy)methyl)propyl1-methylpyrrolidine-2- carboxylate, (2S)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl1- methylpyrrolidine-2-carboxylate, (9Z,9'Z,12Z,12'Z)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)-2-(((9Z,12Z)-octa deca-9,12- dienoyloxy)methyl)propane-1,3-diylbis(octadeca-9,12-dienoate ), (9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((((1-ethylpiperidin-3- yl)methoxy)carbonyl)oxy)methyl)propyloctadeca-9,12-dienoate, 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoy loxy)methyl)propyl1- (cyclopropylmethyl)piperidine-4-carboxylate, 3-((4,4-bis(octyloxy)butanoyl)oxy)-2- (((9Z,12Z)-octadeca-9,12-dienoyloxy)methyl)propyl1-isopropyl piperidine-4-carboxylate, (9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(((3- (dimethylamino)propanoyl)oxy)methyl)propyloctadeca-9,12-dien oate, 4- (dimethylamino)butylcarbonate(6Z,9Z,26Z,29Z)-pentatriaconta- 6,9,26,29-tetraen-18-yl, 3- ((6-(dimethylamino)hexanoyl)oxy)-2,2-bis(((9Z)-tetradec-9-en oyloxy)methyl)propyl(9Z)- tetradec-9-enoate, 2,5-bis((9Z,12Z)-octadeca-9,12-dienyloxy)benzyl3- (dimethylamino)propylcarbonate, (9Z,9'Z,12Z,12'Z)-2-(((4-(pyrrolidin-1- yl)butanoyl)oxy)methyl)propane-1,3-diylbis(octadeca-9,12-die noate), 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl5-heptyldodeca noate, Aceticacid(7R,9Z)- 18-({[3-(dimethylamino)propyloxy]carbonyl}oxy)octacosa-9-en- 7-yl, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl9-pentyltetrad ecanoate, (9Z,12Z)-3-((6,6- bis(octyloxy)hexanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyloctadeca-9,1 2-dienoate, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl7-hexyltridec- 6-enoate, (9Z,12Z)-3-(2,2- bis(heptyloxy)acetoxy)-2-((((2-(dimethylamino)ethoxy)carbony l)oxy)methyl)propyloctadeca- 9,12-dienoate, 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)pentadecyl3-octyl undec-2- enoate, (9Z,12Z)-3-(((3-(diethylamino)propoxy)carbonyl)oxy)-2-(((5- heptyldodecanoyl)oxy)methyl)propyloctadeca-9,12-dienoate, 3-(((3- dimethylamino)propoxy)carbonyl)oxy)pentadecyl3octylundecanoa te, (9Z,12Z)-3-(((3- (diethylamino)propoxy)carbonyl)oxy)-2-(((9- pentyltetradecanoyl)oxy)methyl)propyloctadeca-9,12-dienoate, Diaceticacid(7R,9Z,26Z,29R)-18-({[3- (dimethylamino)propoxy]carbonyl}oxy)pentatriaconta-9,26-dien e-7,29-diyl, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl8,8-bis((2-pro pylpentyl)oxy)octanoate, (9Z,12Z)-3-(((3-(diethylamino)propoxy)carbonyl)oxy)-2-(((7- hexyltridecanoyl)oxy)methyl)propyloctadeca-9,12-dienoate, 3-(((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)pentadecyl8,8-bis(( 2- propylpentyl)oxy)octanoate, (9Z,12Z)-3-(((3-(diethylamino)propoxy)carbonyl)oxy)-2-(((3- octylundecanoyl)oxy)methyl)propyloctadeca-9,12-dienoate, 3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyl8,8-bis((2-prop ylpentyl)oxy)octanoate, 3- (((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl8,8-dibuto xyoctanoate, 3-((5- (dimethylamino)pentanoyl)oxy)-2,2-bis(((9Z)-tetradec-9-enoyl oxy)methyl)propyl(9Z)- tetradec-9-enoate, 3-(Dimethylamino)propylcarbonate(6Z,9Z,26Z,29Z)-pentatriacon tour- 6,9,26,29-tetraen-18-yl, 2,5-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)benzyl3- (dimethylamino)propanoate, (9Z,9'Z,12Z,12'Z)-2-(((3-(4-methylpiperazin-1- yl)propanoyl)oxy)methyl)propane-1,3-diylbis(octadeca-9,12-di enoate), 3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyl8,8-bis(octylox y)octanoate, 3- (Dimethylamino)propyloctacosane-11-ylcarbonate, 2,4-bis((9Z,12Z)-octadeca-9,12- dienyloxy)benzyl4-(dimethylamino)butanoate, (9Z,12Z)-3-(((3- (diethylamino)propoxy)carbonyl)oxy)-2-(((2-heptylundecanoyl) oxy)methyl)propyloctadeca- 9,12-dienoate, 3-(((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl6,6-bis( (2- ethylhexyl)oxy)hexanoate, 2-((((3-(dimethylamino)propoxy)carbonyl)oxy)methyl)propane- 1,3-diylbis(2-heptylundecanoate), 3-(((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl6,6- bis(hexyloxy)hexanoate, 4-methyl-2,5-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)benzyl4 - (dimethylamino)butanoate, 3-(((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl6,6- bis(octyloxy)hexanoate, 4-(dimethylamino)butyl4-methyl-2,5-bis((9Z,12Z)-octadeca-9,1 2- dienyloxy)benzylcarbonate, 3-(((3-(dimethylamino)propoxy)carbonyl)oxy)pendadecyl4,4- bis((2-propylpentyl)oxy)butanoate, 2-(12-dodecyl-3-ethyl-8,14-dioxo-7,9,13-trioxa-3- azaoctadecan-18-yl)propane-1,3-diyldioctanoate, 2-(5-oxo-5-((3-(((3-(piperidin-1- yl)propoxy)carbonyl)oxy)pentadecyl)oxy)pentyl)propane-1,3-di yldioctanoate, 3- (dimethylamino)propyl4-methyl-2,5-bis((9Z,12Z)-octadeca-9,12 -dien-1- yloxy)benzylcarbonate, 3-(((3-(ethyl(methyl)amino)propoxy)carbonyl)oxy)pentadecyl4, 4- bis((2-propylpentyl)oxy)butanoate, 2-(11-dodecyl-3-ethyl-9,15-dioxo-8,10,14-trioxa-3- azanonadecan-19-yl)propane-1,3-diyldioctanoate, 2-(10-dodecyl-3-ethyl-8,15-dioxo-7,9,14- trioxa-3-azanonadecan-19-yl)propane-1,3-diyldioctanoate, 2-(5-((4-((((1-methylpiperidin-4- yl)oxy)carbonyl)oxy)hexadecyl)oxy)-5-oxopentyl)propane-1,3-d iyldioctanoate, 2-(5-((4- ((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)hexadecyl)oxy )-5-oxopentyl)propane-1,3- diyldioctanoate, 2-(5-((4-(((((R)-1-methylpyrrolidin-3-yl)oxy)carbonyl)oxy)he xadecyl)oxy)- 5-oxopentyl)propane-1,3-diyldioctanoate, 2-(5-((4-(((((S)-1-methylpyrrolidin-3- yl)oxy)carbonyl)oxy)hexadecyl)oxy)-5-oxopentyl)propane-1,3-d iyldioctanoate, 2-(5-oxo-5- ((4-(((S)-pyrrolidine-2-carbonyl)oxy)hexadecyl)oxy)pentyl)pr opane-1,3-diyldioctanoate, 2- (5-((4-((1,3-dimethylpyrrolidine-3-carbonyl)oxy)hexadecyl)ox y)-5-oxopentyl)propane-1,3- diyldioctanoate, 2-(5-((4-((1,4-dimethylpiperidine-4-carbonyl)oxy)hexadecyl)o xy)-5- oxopentyl)propane-1,3-diyldioctanoate, 4,4-bis(octyloxy)butyl(3- (diethylamino)propyl)pentadecane-1,3-diyldicarbonate, 3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyl4,4-bis((2-prop ylpentyl)oxy)butanoate, ((2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-1,4-phenyle ne)bis(oxy))bis(octane-8,1- diyl)bis(decanoate), 4,4-bis(octyloxy)butyl5-(((3- (diethylamino)propoxy)carbonyl)oxy)heptadecanoate, 6-((6,6-bis(octyloxy)hexanoyl)oxy)-4- (((3-(diethylamino)propoxy)carbonyl)oxy)hexyloctanoate, (12Z,15Z)-3-(((3- (diethylamino)propoxy)carbonyl)oxy)henicosa-12,15-dien-1-yl6 ,6-bis(octyloxy)hexanoate, 3- (((3-(diethylamino)propoxy)carbonyl)oxy)tridecyl6,6-bis(octy loxy)hexanoate, 3-(((3- (diethylamino)propoxy)carbonyl)oxy)undecyl6,6-bis(octyloxy)h exanoate, 3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyl5-(4,6-diheptyl -1,3-dioxan-2-yl)pentanoate, 3-((5-(diethylamino)pentanoyl)oxy)pentadecyl6,6-bis(octyloxy )hexanoate, 1-((6,6- bis(octyloxy)hexanoyl)oxy)pentadecan-3-yl1,4-dimethylpiperid ine-4-carboxylate, 3-((3-(1- methylpiperidin-4-yl)propanoyl)oxy)pentadecyl6,6-bis(octylox y)hexanoate, 1-((6,6- bis(octyloxy)hexanoyl)oxy)pentadecan-3-yl1,3-dimethylpyrroli dine-3-carboxylate, 3-(((3- (diethylamino)propoxy)carbonyl)oxy)pentadecyl4,4-bis((2-ethy lhexyl)oxy)butanoate, 2- (((1,3-dimethylpyrrolidine-3-carbonyl)oxy)methyl)propane-1,3 -diylbis(8- (octanoyloxy)octanoate), ((2-((((3-(dimethylamino)propoxy)carbonyl)oxy)methyl)-1,4- phenylene)bis(oxy))bis(octane-8,1-diyl)bis(decanoate), (2R)-1-((6,6- bis(octyloxy)hexanoyl)oxy)pentadecan-3-ylpyrrolidine-2-carbo xylate, (2S)-1-((6,6- bis(octyloxy)hexanoyl)oxy)pentadecan-3-yl1-methylpyrrolidine -2-carboxylate, (2R)-1-((6,6- bis(octyloxy)hexanoyl)oxy)pentadecan-3-yl1-methylpyrrolidine -2-carboxylate, 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl6,6-bis((3-eth ylpentyl)oxy)hexanote, 3- (((3-(dimethylamino)propoxy)carbonyl)oxy)pentadecyl6,6-bis(( 2- propylpentyl)oxy)hexanoate, 3-(((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl6,6- bis((2-propylpentyl)oxy)hexanoate, 3-(((2- (diethylamino)ethoxy)carbonyl)oxy)pentadecyl6,6-bis(octyloxy )hexanoate, 3-(((3- morpholinoproproxy)carbonyl)oxy)pentadecyl6,6-bis(octyloxy)h exanoate, 3-((((1- methylpiperidin-4-yl)methoxy)carbonyl)oxy)pentadecyl6,6-bis( octyloxy)hexanoate, 3-(((3- (4-methylpiperazin-1-yl)propoxy)carbonyl)oxy)pentadecyl6,6-b is(octyloxy)hexanoate, 3- (((3-(diethylamino)propoxy)carbonyl)oxy)pentadecyl4,4-bis(oc tyloxy)butanoate, 2-(((4- (dimethylamino)butanoyl)oxy)methyl)-2-((dodecanoyloxy)methyl )propane-1,3- diyl(9Z,9′Z)bis-tetradec-9-enoate, (9Z,9'Z,12Z,12'Z)-2-(((4- (dimethylamino)butanoyl)oxy)methyl)propane-1,3-diylbis(octad eca-9,12-dienoate), 3-(((4- (diethylamino)butoxy)carbonyl)oxy)pentadecyl6,6-bis(octyloxy )hexanote, 3-(((3-(piperazin- 1-yl)propoxy)carbonyl)oxy)pentadecyl6,6-bis(octyloxy)hexanoa te, 3-(((3-piperidin-1- yl)propoxy)carbonyl)oxy)pentadecyl6.6-bis(octyloxy)hexanoate , 3-(((3- (dimethylamino)propoxy)carbonyl)oxy)pentadecyl4,4-bis(octylo xy)butanoate, (9Z,9'Z,12Z,12'Z)-2-(9-dodecyl-2-methyl-7,12-dioxo-6,8,13-tr ioxa-2-azatetradecan-14- yl)propane-1,3-diylbis(octadeca-9,12-dienoate), (9Z,12Z)-10-dodecyl-3-ethyl-14-(2- ((9Z,12Z)-octadeca-9,12-dienoyloxy)ethyl)-8,13-dioxo-7,9-dio xa-3,14-diazahexadecan-16- yloctadeca-9,12-dienoate, 2-((2-(((3- (diethylamino)propoxy)carbonyl)oxy)tetradecanoyl)oxy)propane -1,3-diyldioctanoate, 2-(9- dodecyl-2-methyl-7,13-dioxo-6,8,12-trioxa-2-azanonadecan-19- yl)propane-1,3- diyldioctanoate, 2-((decanoyloxy)methyl)-2-(((4- (dimethylamino)butanoyl)oxy)methyl)propane-1,3-diyl(9Z,9′Z )bis-tetradec-9-enoate, (9Z,9'Z,12Z,12'Z)-2-(((3-morpholinopropanoyl)oxy)methyl)prop ane-1,3-diylbis(octadeca- 9,12-dienoate), 3-(Dimethylamino)propylcarbonate(6Z,9Z,28Z,31Z)-heptatricont a-6,9,28,31- tetraen-19-yl, 2,5-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)benzyl4- (dimethylamino)butanoate, 2-(10-dodecyl-3-ethyl-8,14-dioxo-7,9,13-trioxa-3-azaoctadeca n- 18-yl)propane-1,3-diyldioctanoate, (9Z,9'Z,12Z,12'Z)-2-(((1,3-dimethylpyrrolidine-3- carbonyl)oxy)methyl)propane-1,3-diylbis(octadeca-9,12-dienoa te), ((5- ((dimethylamino)methyl)benzene-1,2,3-triyl)tris(oxy))tris(de cane10,1-diyl)trioctanoate, 0',0- (((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(prop ane-3,1-diyl))9- dioctyldinonanedioate, (9Z,12Z)-3-(3-((dimethylamino)methyl)-5-(3-((3- octylundecanoyl)oxy)propoxy)phenoxy)propyloctadeca-9,12-dien oate, ((((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(propane-3 ,1-diyl))bis(oxy))bis(4- oxobutane-4,1-diyl)bis(decanoate), (R)-4-(3-((R)-3,4-bis(octanoyloxy)butoxy)-5- ((dimethylamino)methyl)phenoxy)butane-1,2-diyldioctanoate, (S)-4-(3-((S)-3,4- bis(octanoyloxv)butoxv)-5-((dimethylamino)methyl)phenoxy)but ane-1,2-diyldioctanoate, (R)-4-(3-((S)-3,4-bis(octanoyloxy)butoxy)-5-((dimethylamino) methyl)phenoxy)butane-1,2- diyldioctanoate, 4,4'-((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis( butane1,2- diyl)tetraoctanoate, didodecyl6,6'-((5-((dimethylamino)methyl)-1,3- phenylene)bis(oxy))dihexanoate, di((9Z,12Z)-octadeca-9,12-dien-1-yl)5,5'-((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))dipentanoate, (((5- ((dimethylamino)methyl)-1,3-phenylene)bis(methylene))bis(oxy ))bis(6-oxohexane-6,1- diyl)bis(decanoate), (5-((dimethylamino)methyl)-1,3-phenylene)bis(methylene)bis(8 - (octanoyloxy)octanoate), (5-((dimethylamino)methyl)-1,3-phenylene)bis(methylene)bis(1 0- (octanoyloxy)decanoate), (((5-((dimethylamino)methyl)-1,3- phenylene)bis(methylene))bis(oxy))bis(6-oxohexane-6,1-diyl)d ioctanoate, (((5- ((dimethylamino)methyl)-1,3-phenylene)bis(methylene))bis(oxy ))bis(8-oxooctane-8,1- diyl)bis(decanoate), (9Z,9'Z,12Z,12'Z)-(((5-((dimethylamino)methyl)-1,3- phenylene)bis(methylene))bis(oxy))bis(4-oxobutane-4,1-diyl)b is(octadeca-9,12-dienoate), 0',0-((5-((dimethylamino)methyl)-1,3-phenylene)bis(methylene ))8-dinonyldioctanedioate, 0,0'-((5-((dimethylamino)methyl)-1,3-phenylene)bis(methylene ))bis(10- (octanoyloxy)decyl)disuccinate, 0,0'-((5-((dimethylamino)methyl)-1,3- phenylene)bis(methylene))di((9Z,12Z)-octadeca-9,12-dien-1-yl )disuccinate, (9Z,9'Z,12Z,12'Z)-(5-((((3-(diethylamino)propoxy)carbonyl)ox y)methyl)-1,3- phenylene)bis(methylene)bis(octadeca-9,12-dienoate), (9Z,12Z)-4-(3- ((dimethylarnino)methyl)-5-(4-(oleoyloxy)butoxy)phenoxy)buty loctadeca-9,12-dienoate, (9Z,9'Z,12Z,12'Z,15Z,15'Z)-((5-((dimethvlamino)methyl)-1,3- phenylene)bis(oxy))bis(butane-4,1-diyl)bis(octadeca-9,12,15- trienoate), ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butane-4, 1-diyl)ditetradecanoate, (Z)- ((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butan e-4,1-diyl)dioleate, ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(hexane-6, 1-diyl)didodecanoate, (9Z,9'Z,12Z,12'Z)-((((5-((diethylamino)methyl)-1,3-phenylene )bis(oxy))bis(ethane-2,1- diyl))bis(oxy))bis(ethane-2,1-diyl)bis(octadeca-9,12-dienoat e), didecyl8,8'-((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))dioctanoate, ((5-((dimethylamino)methyl)- 1,3-phenylene)bis(oxy))bis(propane-3,1-diyl)bis(3-octylundec anoate), (9Z.9'Z.12Z.12'Z)-((5- ((diethvlamino)methvn-2-methvl-1.3-phenylene)bis(oxy))bis(bu tane-4,1-diyl)bis(octadeca- 9,12-dienoate), ((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(octan e-8,1- diyl)didodecanoate, ((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(octan e-8,1- diyl)bis(decanoate), (9Z.9'Z.12Z.12'Z)-((5-((dimethvlarnino)methvn-2-methvl-1.3- phenylene)bis(oxy))bis(butane-4,1-diyl)bis(octadeca-9,12-die noate), (8Z,8'Z)-((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(hexane-bi s(dodec-8-enoate), (9Z,9'Z,12Z,12'Z)-((5-((3-hydroxyazetidin-1-yl)methyl)-1,3-p henylene)bis(oxy))bis(butane- 4,1-diyl)bis(octadeca-9,12-dienoate), ((5-((dimethylamino)methyl)-1,3- phenylene)bis(oxy))bis(hexane-6,1-diyl)dioctanoate, ((5-((dimethylamino)methyl)-1,3- phenylene)bis(oxy))bis(hexane-6,1-diyl)bis(decanoate), (9Z.9'Z.12Z.12'Z)-((5- ((dimethvlamino)methvn-1.3-phenylene)bis(oxy))bis(octane-8,1 -diyl)bis(octadeca-9,12- dienoate), (9Z,9'Z,12Z,12'Z)-((5-((dimethvlamino)methyl)-1,3- phenylene)bis(oxy))bis(hexane-6,1-diyl)bis(octadeca-9,12-die noate), ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(decane-10 ,1-diyl)dihexanoate, ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(decane-10 ,1-diyl)dioctanoate, ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(octane-8, 1-diyl)dioctanoate, ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(octane-8, 1-diyl)dihexanoate, (9Z,9'Z,12Z,12'Z)-((5-((dimethvlamino)methyl)-1,3-phenylene) bis(oxy))bis(ethane-2,1- diyl)bis(octadeca-9,12-dienoate), (9Z,9'Z,12Z,12'Z)-((5-((dimethylamino)methyl)-1,3- phenylene)bis(oxy))bis(propane-3,1-diyl)bis(octadeca-9,12-di enoate), (9Z,9'Z,12Z,12'Z)-((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butane-4, 1-diyl)bis(octadeca-9,12- dienoate), (5-((dimethylamino)methyl)-1,3-phenylene)bis(methylene)ditri decanoate, (9Z,9'Z,12Z,12'Z)-(5-((dimethylamino)methyl)-1,3-phenylene)b is(methylene)bis(octadeca- 9,12-dienoate), (2,6-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)pyridin-4-yl)me thyl3- (dimethylamino)propanoate, (9Z,9'Z,12Z,12'Z)-5-(((3- (dimethylamino)propanoyl)oxy)methyl)-1,3-phenylenebis(octade ca-9,12-dienoate), 1-(3,5- bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)phenyl)-N,Ndimethylm ethanamine, 3,5- bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)benzyl3-(dimethylami no)propanoate, 1-(3,5- bis(4,4-bis(octyloxy)butoxy)phenyl)-N,N-dimethylmethanamine, ((((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butane-4, 1-diyl))bis(oxy))bis(propane- 3,2,1-triyl)tetraoctanoate, ((5-(((4-(dimethylamino)butanoyl)oxy)methyl)-1,3- phenylene)bis(oxy))bis(octane-8,1-diyl)bis(decanoate), ((5-(((3- (dimethylamino)propanoyl)oxy)methyl)-1,3-phenylene)bis(oxy)) bis(octane-8,1- diyl)bis(decanoate), (9Z,9'Z,12Z,12'Z)-((5-(3-morpholinopropyl)-1,3- phenylene)bis(oxy))bis(butane4,1-diyl)bis(octadeca-9,12-dien oate), (9Z,9'Z,12Z,12'Z)-((5-(3- (dimethvlamino)propyl)-1,3-phenylene)bis(oxy))bis(butane-4,1 -diyl)bis(octadeca-9,12- dienoate), (9Z,9'Z,12Z,12'Z)-((5-(3-(piperidin-1-yl)propyl)-1,3- phenylene)bis(oxy))bis(butane-4,1-diyl)bis(octadeca-9,12-die noate), (5- ((dimethylamino)methyl)-1,3-phenylene)bis(methylene)bis(9-pe ntyltetradecanoate), (5- ((dimethylamino)methyl)-1,3-phenylene)bis(methylene)bis(7-he xyltridecanoate), (5- ((dimethylamino)methyl)-1,3-phenylene)bis(methylene)bis(5-he ptyldodecanoate), ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butane-4, 1-diyl)bis(3- octylundecanoate), ((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butan e-4,1- diyl)bis(5-heptyldodecanoate), ((5-((dimethylamino)methyl)-1,3- phenylene)bis(oxy))bis(butane-4,1-diyl)bis(9-pentyltetradeca noate), ((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(butane-4, 1-diyl)bis(7- hexyltridecanoate), (9Z,9'Z,12Z,12'Z)-((5-(pyrrolidin-1-ylmethyl)-1,3- phenylene)bis(oxy))bis(butan4,1-diyl)bis(octadeca-9,12-dieno ate), (((5- ((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(methylene ))bis(propane-3,2,1- triyl)tetraoctanoate, (((5-((dimethylamino)methyl)-1,3-phenylene)bis(oxy))bis(buta ne-4,1- diyl))bis(propane-3,2,1-triyl)tetraoctanoate, (9Z.12Z)-4-(3-((dimethvlamino)methvn-5-(4-((3- octylundecanoyl)oxy)butoxy)phenoxy)butyloctadeca-9,12-dienoa te, bis(1,3- bis(octanoyloxy)propan-2-yl)0,0'-((5-((dimethylamino)methyl) -1,3- phenylene)bis(methylene))disuccinate, (5-((dimethylamino)methyl)-1,3- phenylene)bis(methylene)bis(6-(((nonyloxy)carbonyl)oxy)hexan oate), 2-(3-(4-(5- ((dimethylamino)methyl)-2-methyl-3-((9Z,12Z)-octadeca9,12-di en-1- yloxy)phenoxy)butoxy)-3-oxopropyl)propane-1,3-diyldihexanoat e, 3- ((dimethylamino)methyl)-5-(((8-(octanoyloxy)octanoyl)oxy)met hyl)benzyl3- octylundecanoate, ((5-((diethylamino)methyl)benzene-1,2,3-triyl)tris(oxy))tris (decane-10,1- diyl)trioctanoate, 1-(3,5-bis((Z)-octadec-9-en-1-yloxy)phenyl)-N,N-dimethylmeth anamine, N'-methyl-N',N".N"-tris((2E.6E)-3.7.11-trimethyldodeca-2.6.1 0-trien-1-vnpropane-1,3- diamine, l,17-bis(2-((2-pentylcyclopropyl)methyl)cyclopropyl)heptadec an-9-yl4- (dimethylamino)butanoate, ethyl(7Z)-17-{[4-(dimethylamino)butanoyl]oxy}hexacos-7- enoate, (Z)-methyl6-(2-(dimethylamino)-3-(octadec-9-en-1-yloxy)propo xy)hexanoate, 2- (Didodecylamino)-1-(4-(N-(2-(dinonylamino)ethyl)-N-dodecylgl ycyl)piperazin-1-yl)ethan-1- one, 3-((3-(1-(3-((2-(Dinonylamino)ethyl)(nonyl)amino)propanoyl)p iperidin-4- yl)propyl)(nonyl)amino)propylhexanoate, 3-((3-(4-(3-((2- (Dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-1-yl)-3 - oxopropyl)(nonyl)amino)propylhexanoate, 3-((2-(Dinonylamino)ethyl)(nonyl)amino)-1-(4- (3-(dinonylamino)propyl)piperidin-1-yl)propan-1-one, Pentyl4-((3-(1-(3-((2- (dinonylamino)ethyl)(nonyl)amino)propanoyl)piperidin-4-yl)pr opyl)(nonyl)amino)butano, Pentyl4-((2-(1-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)pipe ridin-4- yl)ethyl)(nonyl)amino)butanoate, Pentyl4-(((1-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)pyrrolidin-3-yl)methyl)(nonyl)amino)butanoate, Pentyl4-((2-(1-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3-yl)ethyl)(no nyl)amino)butanoate, Pentyl4- ((2-(1-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperidin-3- yl)ethyl)(nonyl)amino)butanoate, 2-(Didodecylamino)-1-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-1-yl)ethan-1-one, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-1-(3-(2- (dinonylamino)ethyl)piperidin-1-yl)ethan-1-one, Dipentyl4,4'-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-1-yl)-2-oxoethy l)azanediyl)dibutyrate, Pentyl4-(nonyl(2-(4-(N-nonyl-N-(2-(nonyl(4-oxo-4- (pen1yloxy)buryl)amino)ethyl)glycyl)piperazin-1-yl)-2-oxoeth yl)amino)butanoate, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)-1-(3-((dinonylamino)methyl )pyrrolidin-1-yl)ethan-1- one, 2-((2-(Didodecylamino)ethyl)(dodecyl)amino)-1-(4-(dinonylgly cyl)piperazin-1-yl)ethan- 1-one, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-1-(3-(2-(dinonylamin o)ethyl)pyrrolidin-1- yl)ethan-1-one, Pentyl4-((3-(4-(3-((2- (dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-1-yl)-3 - oxopropyl)(nonyl)amino)butanoate, 3-((2-(1-(N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)piperidin-4-yl)ethyl)(nonyl)amino)propylhexanoat e, Butyl5-((2-(1-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)pentanoate, 2-((2- (Didodecylamino)ethyl)(nonyl)amino)-1-(4-(dinonylglycyl)pipe razin-1-yl)ethan-1-one, Propyl6-((2-(1-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)pipe ridin-4- yl)ethyl)(nonyl)amino)hexanoate, Ethyl7-((2-(1-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperidin-4-yl)ethyl)(nonyl)amino)heptanoate, Methyl8-((2-(1-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)octanoate, 3-((2-(4- (N-(2-(Dinonylamino)ethyl)-N-nonylglycyl)piperazin-1-yl)-2- oxoethyl)(nonyl)amino)propylhexanoate, Butyl5-((2-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-1-yl)-2-oxoethyl)(nonyl)amino)pentanoa te, Propyl6-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-2-oxoethyl)(non yl)amino)hexanoate, Ethyl7- ((2-(4-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperazin-1- yl)-2- oxoethyl)(nonyl)amino)heptanoate, 3-(Dinonylamino)-1-(4-(3-((2- (dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-1-yl)pr opan-1-one, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)-1-(4-(ditetradecylglycyl)p iperazin-1-yl)ethan-1-one, 2- (Dinonylamino)-1-(4-(2-((2-(dinonylamino)ethyl)(nonyl)amino) ethyl)piperidin-1-yl)ethan-1- one, 2-(Dinonylamino)-l-(4-(N-(2-(dinonylamino)ethyl)-N-dodecylgl ycyl)piperazin-1- yl)ethan-1-one, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-1-(4-(2- (dinonylamino)ethyl)piperidin-1-yl)ethan-1-one, Methyl8-((2-(4-(dinonylglycyl)piperazin-1- yl)-2-oxoethyl)(2-((8-methoxy-8-oxooctyl)(nonyl)amino)ethyl) amino)octanoate, Methyl8- ((2-(dinonylamino)ethyl)(2-(4-(dinonylglycyl)piperazin-1-yl) -2-oxoethyl)amino)octanoate, Methyl8-((2-((2-(4-(dinonylglycyl)piperazin-1-yl)-2- oxoethyl)(nonyl)amino)ethyl)(nonyl)amino)octanoate, Pentyl4-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-2-oxoethyl)(non yl)amino)butanoate, Methyl8- ((2-(4-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperazin-1- yl)-2- oxoethyl)(nonyl)amino)octanoate, 2-((2-(Didodecylamino)ethyl)(dodecyl)amino)-1-(5- (dinonylglycyl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)ethan-1-o ne3, 2-(Dinonylamino)-1-(5-(N- (2-(dinonylamino)ethyl)-N-nonylglycyl)-2,5-diazabicyclo[2.2. 1]heptan-2-yl)ethan-1-one, N1,N1,N2-Tri((9Z,12Z)-octadeca-9,12-dien-1-yl)-N2-(2-(pipera zin-1-yl)ethyl)ethane-1,2- diamine, N1,N1,N2-Tri((Z)-octadec-9-en-1-yl)-N2-(2-(piperazin-1-yl)et hyl)ethane-1,2- diamine, 2-(Dinonylamino)-l-(4-(N-(2-(dinonylamino)ethyl)-N-nonylglyc yl)piperazin-l- yl)ethan-l-one, N1,N1,N2-Tridodecyl-N2-(2-(piperazin-1-yl)ethyl)ethane-1,2-d iamine, N1,N1,N2-Trinonyl-N2-(2-(piperazin-1-yl)ethyl)ethane-1,2-dia mine, N1,N1,N2-Trihexyl- N2-(2-(piperazin-1-yl)ethyl)ethane-1,2-diamine, N1-(2-(4-(2- (Didodecylamino)ethyl)piperazin-1-yl)ethyl)-N1,N2,N2-tri((9Z ,12Z)-octadeca-9,12-dien-1- yl)ethane-1,2-diamine, N1-(2-(4-(2-(Didodecylamino)ethyl)piperazin-1-yl)ethyl)-N1,N 2,N2- tri((Z)-octadec-9-en-1-yl)ethane-1,2-diamine, N1-(2-(4-(2- (Ditetradecylamino)ethyl)piperazin-1-yl)ethyl)-N1,N2,N2-trit etradecylethane-1,2-diamine, N1-(2-(4-(2-(Didodecylamino)ethyl)piperazin-1-yl)ethyl)-N1,N 2,N2-tritetradecylethane-1,2- diamine, N1-(2-(4-(2-(Dinonylamino)ethyl)piperazin-1-yl)ethyl)-N1,N2, N2- tritetradecylethane-1,2-diamine, 2-(Didodecylamino)-l-(4-(2-((2- (didodecylamino)ethyl)(dodecyl)amino)ethyl)piperazin-1-yl)et han-1-one, N1-(2-(4-(2- (Di((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)ethyl)piperazin-1 -yl)ethyl)-N1,N2,N2- tridodecylethane-1,2-diamine, N1-(2-(4-(2-(Di((Z)-octadec-9-en-1-yl)amino)ethyl)piperazin- 1-yl)ethyl)-N1,N2,N2-tridodecylethane-1,2-diamine, N1,N1,N2-Tridodecyl-N2-(2-(4-(2- (dodecyl((9Z,12Z)-octadeca-9,12-dien-1-yl)amino)ethyl)pipera zin-1-yl)ethyl)ethane-1,2- diamine, N1-(2-(4-(2-(Ditetradecylamino)ethyl)piperazin-1-yl)ethyl)-N 1,N2,N2- tridodecylethane-1,2-diamine, N1-(2-(4-(2-(Di((Z)-dodec-6-en-l-yl)amino)ethyl)piperazin-l- yl)ethyl)-N1,N2,N^tridodecylethane-1,2-diamine, (Z)-N1-(2-(4-(2-(Dodec-6-en-l- yl(dodecyl)amino)ethyl)piperazin-l-yl)ethyl)-N,N2,N2-tridode cylethane-1,2-diamine, N1-(2- (4-(2-(Dinonylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-tri dodecylethane-1,2-diamine, N1-(2-(4-(2-(Dioctylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2, N2-tridodecylethane-1,2- diamine, N1-(2-(4-(2-(Dihexylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2, N2-tridodecylethan- 1,2-diamine, N1-(2-(4-(2-(Ditetradecylamino)ethyl)piperazin-l-yl)ethyl)-N 1,N2,N2- trinonylethane-1,2-diamine, 2-((2-(Didodecylamino)ethyl)(dodecyl)amino)-l-(4-(2- (didodecylamino)ethyl)piperazin-l-yl)ethan-l-one, N1-(2-(4-(2- (Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-trinony lethane-1,2-diamine, N1-(2- (4-(2-(Dinonylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-tri nonylethane-1,2-diamine, N1- (2-(4-(2-(Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N 2-trihexylethane-1,2-diamine, Dimethyl12,12'-((2-(4-(2-((2-(didodecylamino)ethyl)(dodecyl) amino)ethyl)piperazin-l- yl)ethyl)azanediyl)didodecanoate, Methyl12-((2-(4-(2-((2- (didodecylamino)ethyl)(dodecyl)amino)ethyl)piperazin-l- yl)ethyl)(dodecyl)amino)dodecanoate, Dipentyl6,6'-((2-(4-(2-((2- (didodecylamino)ethyl)(dodecyl)amino)ethyl)piperazin-l-yl)et hyl)azanediyl)dihexanoate, Pentyl6-((2-(4-(2-((2-(ditetradecylamino)ethyl)(tetradecyl)a mino)ethyl)piperazin-1- yl)ethyl)(dodecyl)amino)hexanoate, Pentyl6-((2-(4-(2-((2- (didodecylamino)ethyl)(dodecyl)amino)ethyl)piperazin-l-yl)et hyl)(dodecyl)amino)hexanoate, 2-(Didodecylamino)-l-(4-(N-(2-(didodecylamino)ethyl)-N-dodec ylglycyl)piperazin-1- yl)ethan-1-one, 2-(Didodecylamino)-1-(4-(N-(2-(didodecylamino)ethyl)-N- nonylglycyl)piperazin-1-yl)ethan-1-one, 2-(Didodecylamino)-N-(2-(4-(2- (didodecylamino)ethyl)piperazin-l-yl)ethyl)-N-dodecylacetami de, ((2-((3,S',4R)-3,4- dihydroxypyrrolidin-l-yl)acetyl)azanediyl)bis(ethane-2,1-diy l)(9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate), 2-amino-N,N-dihexadecyl-3-(1H-imidazol-5-yl)propanamide, (2-amino-N,N-dihexadecyl-3-(1H-imidazol-5-yl)propanamide, methyl(9Z)-19-[2- (dimethylamino)ethyl]heptacos-9-enoate, methyl8-(2-{9-[2- (dimethylamino)ethyl]octadecyl}cyclopropyl)octanoate, methyl(9Z)-19-[2- (dimethylamino)ethyl]octacos-9-enoate, ethyl8-(2-{ll- [(dimethylamino)methyl]heptadecyl}cyclopropyl)octanoate, ethyl8-(2-{ll- [(dimethylamino)methyl]octadecyl}cyclopropyl)octanoate, di((9Z,12Z)-octadeca-9,12-dien-l- yl)3-(((2-(dimethylamino)ethoxy)carbonyl)amino)pentanedioate , Heptyl6-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(tet radecyl)amino)hexanoate, ethyl8-(2-{ll-[(dimethylamino)methyl]nonadecyl}cyclopropyl)o ctanoate, Pentyl8-((2-(l-(N- (2-(dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)( tetradecyl)amino)octanoate, ethyl8-(2-{ll-[(dimethylamino)methyl]icosyl}cyclopropyl)octa noate, ethyl8-(2-{9- [(dimethylamino)methyl]pentadecyl}cyclopropyl)octanoate, 3-((2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperidin-4- yl)ethyl)(tetradecyl)amino)propyldecanoate, Heptyl6-((2-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(tetradecyl)amino)hex anoate, ethyl8-(2-{9- [(dimethylamino)methyl]hexadecyl}cyclopropyl)octanoate, Pentyl8-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-2-oxoethyl)(tet radecyl)amino)octanoate, ethyl8-(2-{9-[(dimethylamino)methyl]heptadecyl}cyclopropyl)o ctanoate, methyl6-(2-(8-(2- (dimethylamino)-3-(nonyloxy)propoxy)octyl)cyclopropyl)hexano ate, methyl(9Z)-21- (dimethylamino)heptacos-9-enoate, methyl(9Z)-21-{[4- (dimethylamino)butanoyl]oxy}heptacos-9-enoate, (2R)-N,N-dimethyl-1-[(9Z,12Z)-octadeca- 9,12-dien-1-yloxy]dodecan-2-amine, (15Ζ,18Ζ)-Ν,Ν-dimethyltetracoda-15,18-dien-5-amine, ethyl8-(2-{9-[(dimethylamino)methyl]octadecyl}cyclopropyl)oc tanoate, 3-((2-(4-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2- oxoethyl)(tetradecyl)amino)propyldecanoate, ethyl4-(2-{ll- [(dimethylamino)methyl]icosyl}cyclopropyl)butanoate, ethyl8-(2-{7- [(dimethylamino)methyl]hexadecyl}cyclopropyl)octanoate, 3-((3-(l-(3-((2- (Dinonylamino)ethyl)(nonyl)amino)propanoyl)piperidin-4- yl)propyl)(nonyl)amino)propylhexanoate, ethyl6-(2-{9- [(dimethylamino)methyl]pentadecyl}cyclopropyl)hexanoate, 3-((3-(4-(3-((2- (Dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-l-yl)-3 - oxopropyl)(nonyl)amino)propylhexanoate, ethyl6-(2-{9- [(dimethylamino)methyl]hexadecyl}cyclopropyl)hexanoate, 3-((2- (Dinonylamino)ethyl)(nonyl)amino)-l-(4-(3-(dinonylamino)prop yl)piperidin-1-yl)propan-1- one, Pentyl4-((3-(l-(3-((2-(dinonylamino)ethyl)(nonyl)amino)propa noyl)piperidin-4- yl)propyl)(nonyl)amino)buta^, ethyl6-(2-{9- [(dimethylamino)methyl]heptadecyl}cyclopropyl)hexanoate, Pentyl4-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)butanoate, ethyl6-(2- {9-[(dimethylamino)methyl]octadecyl}cyclopropyl)hexanoate, Pentyl4-(((l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3-yl)methyl)(n onyl)amino)butanoate, ethyl(9Z)-21-[(dimethylamino)methyl]heptacos-9-enoate, Pentyl4-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3-yl)ethyl)(no nyl)amino)butanoate, ethyl(9Z)-21-[(dimethylamino)methyl]octacos-9-enoate, ((2-((3,S',4R)-3,4- dihydroxypyrrolidin-l-yl)acetyl)azanediyl)bis(ethane-2,l-diy l)(9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate), Pentyl4-((2-(l-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperidin-3-yl)ethyl)(nonyl)amino)butanoate, ethyl(9Z)-21- [(dimethylamino)methyl]nonacos-9-enoate, methyl6-(2-(8-(2-(dimethylamino)-3- (heptyloxy)propoxy)octyl)cyclopropyl)hexanoate, methyl(9Z)-21-{[4- (dimethylamino)butanoyl]oxy}octacos-9-enoate, methyl(9Z)-21-(dimethylamino)octacos-9- enoate, 2-(Didodecylamino)-1-(4-(N-(2-(dinonylamino)ethyl)-N-nonylgl ycyl)piperazin-1- yl)ethan-1-, (2S)-N.N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]non an-2-amine, (18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-10-amine, ethyl(9Z)-21- [(dimethylamino)methyl]triacont-9-enoate, ethyl(9Z)-19-[(dimethylamino)methyl]pentacos- 9-enoate, ethyl(9Z)-19-[(dimethylamino)methyl]hexacos-9-enoate, ethyl(9Z)-19- [(dimethylamino)methyl]heptacos-9-enoate, ethyl(9Z)-19-[(dimethylamino)methyl]octacos- 9-enoate, ethyl(5Z)-17-[(dimethylamino)methyl]hexacos-5-enoate, ethyl(9Z)-17- [(dimethylamino)methyl]hexacos-9-enoate, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-l-(3- (2-(dinonylamino)ethyl)piperidin-l-yl)ethan-l-one, ethyl(7Z)-17- [(dimethylamino)methyl]tricos-7-enoate, Dipentyl4,4'-((2-(4-(N-(2-(dinonylarnino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)azanediyl)dibutyrate, Pentyl4-(nonyl(2-(4-(N-nonyl- N-(2-(nonyl(4-oxo-4-(pentyloxy)butyl)amino)ethyl)glycyl)pipe razin-l-yl)-2- oxoethyl)amino)butanoate, ethyl(7Z)-17-[(dimethylamino)methyl]tetracos-7-enoate, ethyl(7Z)-17-[(dimethylamino)methyl]pentacos-7-enoate, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)-l-(3-((dinonylamino)methyl )pyrrolidin-1-yl)ethan-1- one, trans-3-[(3}7-dimethyloctyl)oxy]-1-methyl-4~[(9Z,12Z)-octade ca-9512-dien-1- yloxyjpyrrolidine, methyl6-(2-(8-(2-(dimethylamino)-3- (hexyloxy)propoxy)octyl)cyclopropyl)hexanoate, methyl(9Z)-21-{[4- (dimethylamino)butanoyl]oxy}nonacos-9-enoate, methyl(9Z)-21-(dimethylamino)nonacos-9- enoate, (2S)-N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]tri decan-2-amine, (15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-7-amine, ethyl(7Z)-17- [(dimethylamino)methyl]hexacos-7-enoate, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-l-(3- (2-(dinonylamino)ethyl)pyrrolidin-1-yl)ethan-1-one, methyl6-(2-{ll- [(dimethylamino)methyl]icosyl}cyclopropyl)hexanoate, methyl10-(2-{7- [(dimethylamino)methyl]hexadecyl}cyclopropyl)decanoate, methyl8-(2-{ll- [(dimethylamino)methyl]heptadecyl}cyclopropyl)octanoate, methyl8-(2-{ll- [(dimethylamino)methyl]octadecyl}cyclopropyl)octanoate, methyl8-(2-{ll- [(dimethylamino)methyl]nonadecyl}cyclopropyl)octanoate, methyl8-(2-{ll- [(dimethylamino)methyl]icosyl}cyclopropyl)octanoate, Pentyl4-((3-(4-(3-((2- (dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-l-yl)-3 - oxopropyl)(nonyl)amino)butanoate, methyl8-(2-{9- [(dimethylamino)methyl]pentadecyl}cyclopropyl)octanoate, methyl8-(2-{9- [(dimethylamino)methyl]hexadecyl}cyclopropyl)octanoate, 3-((2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)propylhexanoate, methyl8-(2-{9-[(dimethylamino)methyl]heptadecyl}cyclopropyl) octanoate, methyl8-(2- (dimethylamino)-3-((6-((2-octylcyclopropyl)methoxy)-6-oxohex yl)oxy)propoxy)octanoate, Butyl5-((2-(l-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piper idin-4- yl)ethyl)(nonyl)amino)pentanoate, trans-1-methyl-3-[(12Z)-octadec-12-en-1-yloxy]-4- (octyloxy)pyrrolidine, methyl(9Z)-21-{[4-(dimethylamino)butanoyl]oxy}triacont-9-eno ate, methyl(9Z)-21-(dimethylamino)triacont-9-enoate, 2-((2- (Didodecylamino)ethyl)(nonyl)amino)-1-(4-(dinonylglycyl)pipe razin-1-yl)ethan-1- oneStep1:MethylN-(2-(didodecylamino)ethyl)-N-nonylglycinate, 1-((2R,3S,5R)-3- (bis(hexadecyloxy)methoxy)-5-(5-methyl-2,4-dioxo-3,4-dihydro pyrimidin-1(2H)- yl)tetrahydrofumethanesulfonate, (Z)-methyl16-(3-(decyloxy)-2- (dimethylamino)propoxy)hexadec-7-enoate, (2S)-1-[(9Z,12Z)-octadeca-9,12-dien-1- yloxy]nonan-2-amine, (14Z,17Z)-N,N-dimethyltricosa-14,17-dien-6-amine, Propyl6-((2-(l- (N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethy l)(nonyl)amino)hexanoate, methyl7-(2-(dimethylamino)-3-((6-((2-octylcyclopropyl)methox y)-6- oxohexyl)oxy)propoxy)heptanoate, methyl(7Z)-19-[(dimethylamino)methyl]octacos-7- enoate, methyl(HZ)-19-[(dimethylamino)methyl]octacos-ll-enoate, Ethyl7-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)heptanoate, (2- octylcyclopropyl)methyl6-(2-(dimethylamino)-3-((5-methoxy-5- oxopentyl)oxy)propoxy)hexanoate, Methyl8-((2-(l-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperidin-4-yl)ethyl)(nonyl)amino)octanoate, methyl(9Z)-21- [(dimethylamino)methyl]heptacos-9-enoate, (2-octylcyclopropyl)methyl6-(2- (dimethylamino)-3-(4-methoxy-4-oxobutoxy)propoxy)hexanoate, methyl(9Z)-21- [(dimethylamino)methyl]octacos-9-enoate, 3-((2-(4-(N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(nonyl)amino)propylhe xanoate, (Z)-methyl8-(2- (dimethylamino)-3-((6-oxo-6-(undec-2-en-l-yloxy)hexyl)oxy)pr opoxy)octanoate, methyl(9Z)-21-[(dimethylamino)methyl]nonacos-9-enoate, Butyl5-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoethy l)(nonyl)amino)pentanoate, (Z)-methyl7-(2-(dimethylamino)-3-((6-oxo-6-(undec-2-en-l- yloxy)hexyl)oxy)propoxy)heptanoate, Propyl6-((2-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(nonyl)amino)hexanoat e, methyl(9Z)-21- [(dimethylamino)methyl]triacont-9-enoate, (Z)-undec-2-en-1-yl6-(2-(dimethylamino)-3-((5- methoxy-5-oxopentyl)oxy)propoxy)hexanoate, methyl(9Z)-19- [(dimethylamino)methyl]pentacos-9-enoate, Ethyl7-((2-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(nonyl)amino)heptanoa te, (Z)-undec-2-en-1-yl6-(2- (dimethylamino)-3-(4-methoxy-4-oxobutoxy)propoxy)hexanoate, methyl6-(2- (dimethylamino)-3-((6-((2-octylcyclopropyl)methoxy)-6-oxohex yl)oxy)propoxy)hexanoate, methyl(9Z)-19-[(dimethylamino)methyl]hexacos-9-enoate, 3-(Dinonylamino)-l-(4-(3-((2- (dinonylamino)ethyl)(nonyl)amino)propanoyl)piperazin-1-yl)pr opan-1-one, methyl(9Z)-19- [(dimethylamino)methyl]heptacos-9-enoate, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-l-(4- (ditetradecylglycyl)piperazin-1-yl)ethan-1-one, (Z)-methyl6-(2-(dimethylamino)-3-((6-oxo-6- (undec-2-en-l-yloxy)hexyl)oxy)propoxy)hexanoate, methyl8-(2-(dimethylamino)-3-((8-(2-(6- methoxy-6-oxohexyl)cyclopropyl)octyl)oxy)propoxy)octanoate, methyl8-(2-{9- [(dimethylamino)methyl]octadecyl}cyclopropyl)octanoate, 2-(Dinonylamino)-l-(4-(2-((2- (dinonylamino)ethyl)(nonyl)amino)ethyl)piperidin-1-yl)ethan- 1-one, trans-l-methyl-3-[(9Z)- octadec-9-en-l-yloxy]-4-(octyloxy)pyrrolidine, methyl(9Z)-19-{[4- (dimethylamino)butanoyl]oxy}pentacos-9-enoate, methyl(9Z)-19-(dimethylamino)pentacos- 9-enoate, (Z)-methyl16-(2-(dimethylamino)-3-(nonyloxy)propoxy)hexadec- 7-enoate, (2S)-1- [(9Z,12Z)-octadeca-9,12-dien-1-yloxy]decan-2-amine, (12Z,15Z)-N,N-dimethylhenicosa- 12,15-dien-4-amine, methyl7-(2-(dimethylamino)-3-((8-(2-(6-methoxy-6- oxohexyl)cyclopropyl)octyl)oxy)propoxy)heptanoate, methyl(9Z)-19- [(dimethylamino)methyl]octacos-9-enoate, 2-((2-(Dinonylamino)ethyl)(nonyl)amino)-l-(4-(2- (dinonylamino)ethyl)piperidin-l-yl)ethan-l-one, Methyl8-((2-(4-(dinonylglycyl)piperazin-l- yl)-2-oxoethyl)(2-((8-methoxy-8-oxooctyl)(nonyl)amino)ethyl) amino)octanoate, methyl6-(2- (8-(2-(dimethylamino)-3-((5-methoxy-5- oxopentyl)oxy)propoxy)octyl)cyclopropyl)hexanoate, ethyl8-{2-[ll- (dimethylamino)heptadecyl]cyclopropyl}octanoate, Methyl8-((2-(dinonylamino)ethyl)(2-(4- (dinonylglycyl)piperazin-l-yl)-2-oxoethyl)amino)octanoate, methyl6-(2-(8-(2- (dimethylamino)-3-(4-methoxy-4-oxobutoxy)propoxy)octyl)cyclo propyl)hexanoate, ethyl8- {2-[ll-(dimethylamino)octadecyl]cyclopropyl}octanoate, Methyl8-((2-((2-(4- (dinonylglycyl)piperazin-l-yl)-2-oxoethyl)(nonyl)amino)ethyl )(nonyl)amino)octanoate, ethyl8-{2-[ll-(dimethylamino)nonadecyl]cyclopropyl}octanoate , (Z)-methyl16-(2- (dimethylamino)-3-((8-methoxy-8-oxooctyl)oxy)propoxy)hexadec -7-enoate, Pentyl4-((2-(4- (N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2- oxoethyl)(nonyl)amino)butanoate, ethyl8-{2-[ll- (dimethylamino)icosyl]cyclopropyl}octanoate, (Z)-methyl16-(2-(dimethylamino)-3-((7- methoxy-7-oxoheptyl)oxy)propoxy)hexadec-7-enoate, Methyl8-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoethy l)(nonyl)amino)octanoate, ethyl8-{2-[9-(dimethylamino)pentadecyl]cyclopropyl}octanoate , (Z)-methyl16-(2- (dimethylamino)-3-((5-methoxy-5-oxopentyl)oxy)propoxy)hexade c-7-enoate, (11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,23-trien-10-amine, N,N-dimethyl-1-[(1S,2R)-2- octylcyclopropyl]pentadecan-8-amine, ethyl8-{2-[9- (dimethylamino)hexadecyl]cyclopropyl}octanoate, 2-((2- (Didodecylamino)ethyl)(dodecyl)amino)-l-(5-(dinonylglycyl)-2 ,5-diazabicyclo[2.2.1]heptan- 2-yl)ethan-1-one3, (Z)-methyl16-(2-(dimethylamino)-3-(4-methoxy-4- oxobutoxy)propoxy)hexadec-7-enoate, methyl6-(2-(8-(2-(dimethylamino)-3-((6-methoxy-6- oxohexyl)oxy)propoxy)octyl)cyclopropyl)hexanoate, ethyl8-{2-[9- (dimethylamino)heptadecyl]cyclopropyl}octanoate, 2-(Dinonylamino)-l-(5-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)-2,5-diazabicyclo[2.2.1]h eptan-2-yl)ethan-l-one, 1- [(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine, N1,N1,N2-Tri((9Z,12Z)- octadeca-9,12-dien-l-yl)-N2-(2-(piperazin-l-yl)ethyl)ethane- 1,2-diamine, ethyl8-{2-[9- (dimethylamino)octadecyl]cyclopropyl}octanoate, 1-[(1R,2S)-2-heptylcyclopropyl]-Ν,Ν- dimethyloctadecan-9-amine, (Z)-methyl16-(2-(dimethylamino)-3-((6-methoxy-6- oxohexyl)oxy)propoxy)hexadec-7-enoate, N1,N1,N2-Tri((Z)-octadec-9-en-l-yl)-N2-(2- (piperazin-l-yl)ethyl)ethane-l,2-diamine, N,N-dimethyl-3-{7-[(1S,2R)-2- octylcyclopropyl]heptyl}dodecan-1-amine, methyl8-(2-(dimethylamino)-3-((8-(2-((2- pentylcyclopropyl)methyl)cyclopropyl)octyl)oxy)propoxy)octan oate, ethyl4-{2-[ll- (dimethylamino)icosyl]cyclopropyl}butanoate, trans-1-Methyl-3-[((9Z,12Z)-octadeca-9,12- dienyl)oxy]-4-octyloxy-pyrrolidine, methyl(9Z)-19-(dimethylamino)hexacos-9-enoate, methyl(9Z)-19-{[4-(dimethylamino)butanoyl]oxy}hexacos-9-enoa te, (Z)-methyl16-(2- (dimethylamino)-3-(heptyloxy)propoxy)hexadec-7-enoate, (2R)-1-[(9Z,12Z)-octadeca-9,12- dien-1-yloxy]dodecan-2-amine, (13Z,16Z)-N,N-dimethyldocosa-13,16-dien-5-amine, Ν,Ν- dimethyl-1-[(1R,2S)-2-undecylcyclopropyl]tetradecan-5-amine, methyl7-(2- (dimethylamino)-3-((8-(2-((2- pentylcyclopropyl)methyl)cyclopropyl)octyl)oxy)propoxy)hepta noate, ethyl8-{2-[7- (dimethylamino)hexadecyl]cyclopropyl}octanoate, 2-(Didodecylamino)-N-dodecyl-N-(2- (piperazin-l-yl)ethyl)acetamide, Ν,Ν-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8- amine, N1-(2-(Piperazin-l-yl)ethyl)-N1,N2,N2-tritetradecylethane-l, 2-diamine, methyl6-(2- (dimethylamino)-3-((8-(2-((2- pentylcyclopropyl)methyl)cyclopropyl)octyl)oxy)propoxy)hexan oate, ethyl6-{2-[9- (dimethylamino)pentadecyl]cyclopropyl}hexanoate, Ν,Ν-dimethyl-1-[(1S,2S)-2-{[(1R,2R)- 2-pentylcyclopropyl]methyl}cyclopropyl]nonadecan-10-amine, NN1,N2-Tridodecyl-N2-(2- (piperazin-l-yl)ethyl)ethane-l,2-diamine, methyl5-(2-(dimethylamino)-3-((8-(2-((2- pentylcyclopropyl)methyl)cyclopropyl)octyl)oxy)propoxy)penta noate, ethyl6-{2-[9- (dimethylamino)hexadecyl]cyclopropyl}hexanoate, N,N-dimethyl-21-[(1S,2R)-2- octylcyclopropyl]henicosan-10-amine, NNN2-Trinonyl-N2-(2-(piperazin-l-yl)ethyl)ethane- l,2-diamine, methyl4-(2-(dimethylamino)-3-((8-(2-((2- pentylcyclopropyl)methyl)cyclopropyl)octyl)oxy)propoxy)butan oate, ethyl6-{2-[9- (dimethylamino)heptadecyl]cyclopropyl}hexanoate, Ν,Ν-dimethyl-1-[(1S,2R)-2- octylcyclopropyl]nonadecan-10-amine, N1,N1,N2-Trihexyl-N2-(2-(piperazin-l- yl)ethyl)ethane-l,2-diamine, methyl8-(2-(dimethylamino)-3-((9Z,12Z)-octadeca-9,12-dien-l- yloxy)propoxy)octanoate, ethyl6-{2-[9-(dimethylamino)octadecyl]cyclopropyl}hexanoate, N1-(2-(4-(2-(Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N 2,N2-tri((9Z,12Z)-octadeca- 9,12-dien-l-yl)ethane-1,2-diamine, methyl7-(2-(dimethylamino)-3-((9Z,12Z)-octadeca-9,12- dien-1-yloxy)propoxy)heptanoate, ethyl(9Z)-21-(dimethylamino)heptacos-9-enoate, 1- [(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine, 1-methyl18-[(2Z)-non-2- en-1-yl]9-{[4-(dimethylamino)butanoyl]oxy}octadecanedioate, N1-(2-(4-(2- (Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-tri((Z) -octadec-9-en-l-yl)ethane-l,2- diamine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amin e, methyl6-(2- (dimethylamino)-3-((9Z,12Z)-octadeca-9,12-dien-1-yloxy)propo xy)hexanoate, ethyl(9Z)-21- (dimethylamino)octacos-9-enoate, dimethyl(9Z)-19-{[4- (dimethylamino)butanoyl]oxy}heptacos-9-enedioate, N1-(2-(4-(2- (Ditetradecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-trit etradecylethane-1,2-diamine, methyl5-(2-(dimethylamino)-3-((9Z,12Z)-octadeca-9,l2-dien-l- yloxy)propoxy)pentanoate, ethyl8-{[4-(dimethylamino)butanoyl]oxy}-15-(2-octylcycloprop yl)pentadecanoate, ethyl(9Z)-21-(dimethylamino)nonacos-9-enoate, (13Z,16Z)-N,N-dimethyl-3-nonyldocosa- 13,16-dien-1-amine, N1-(2-(4-(2-(Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N 2,N2- tritetradecylethane-1,2-diamine, methyl9-{[4-(dimethylamino)butanoyl]oxy}-16-(2- octylcyclopropyl)hexadecanoate, methyl4-(2-(dimethylamino)-3-((9Z,12Z)-octadeca-9,12- dien-1-yloxy)propoxy)butanoate, ethyl(9Z)-21-(dimethylamino)triacont-9-enoate, (12Z,15Z)- N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine, methyl8-(2-(dimethylamino)-3-((8-(2- octylcyclopropyl)octyl)oxy)propoxy)octanoate, ethyl(9Z)-19-(dimethylamino)pentacos-9- enoate, ethyl(18Z,21Z)-8-{[4-(dimethylamino)butanoyl]oxy}heptacosa-1 8,21-dienoate, (16Z)-N,N-dimethylpentacos-16-en-8-amine, methyl(9Z)-19-{[4- (dimethylamino)butanoyl]oxy}heptacos-9-enoate, methyl(9Z)-19-(dimethylamino)heptacos- 9-enoate, 2-(Didodecylamino)-l-(4-(2-((2- (didodecylamino)ethyl)(dodecyl)amino)ethyl)piperazin-l-yl)et han-l-one, (Z)-methyl16-(2- (dimethylamino)-3-(hexyloxy)propoxy)hexadec-7-enoate, (2S)-1-[(9Z,12Z)-octadeca-9,12- dien-1-yloxy]dodecan-2-amine, (16Z,19Z)-N,N-dimethylpentacosa~16,19-dien-8-amine, N1- (2-(4-(2-(Dinonylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2^V2- tritetradecylethane-l,2- diamine, methyl7-(2-(dimethylamino)-3-((8-(2- octylcyclopropyl)octyl)oxy)propoxy)heptanoate, methyl(19Z,22Z)-9-{[4- (dimethylamino)butanoyl]oxy}octacosa-19,22-dienoate, ethyl(9Z)-19- (dimethylamino)hexacos-9-enoate, (22Z)-N,N-dimethylhentriacont-22-en-10-amine, N1-(2- (4-(2-(Di((Z)-octadec-9-en-l-yl)amino)ethyl)piperazin-l-yl)e thyl)- !^^-tridodecylethane-1,2- diamine, methyl5-(2-(dimethylamino)-3-((8-(2- octylcyclopropyl)octyl)oxy)propoxy)pentanoate, ethyl(9Z)-19-(dimethylamino)heptacos-9- enoate, (2-butylcyclopropyl)methyl12-{[4-(dimethylamino)butanoyl]oxy }henicosanoate, (20Z)-N,N-dimethylnonacos-20-en-10-amine, N1,N1,N2-Tridodecyl-N2-(2-(4-(2- (dodecyl((9Z,12Z)-octadeca-9,12-dien--yl)amino)ethyl)piperaz in-1-yl)ethyl)ethane-1,2- diamine, methyl4-(2-(dimethylamino)-3-((8-(2- octylcyclopropyl)octyl)oxy)propoxy)butanoate, ethyl(9Z)-19-(dimethylamino)octacos-9- enoate, (2-octylcyclopropyl)methyl8-{[4-(dimethylamino)butanoyl]oxy} heptadecanoate, (24Z)-N,N-dimethyltritriacont-24-en-10-amine, N1-(2-(4-(2- (Ditetradecylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2,N2-trid odecylethane-1,2-diamine, ethyl(5Z)-17-(dimethylamino)hexacos-5-enoate, (Z)-methyl8-(2-(dimethylamino)-3-(octadec- 9-en-l-yloxy)propoxy)octanoate, (2Z)-hept-2-en-l-yl12-{[4- (dimethylamino)butanoyl]oxy}henicosanoate, (17Z)-N,N-dimethylnonacos-17-en-10-amine, N1-(2-(4-(2-(Di((Z)-dodec-6-en-l-yl)amino)ethyl)piperazin-l- yl)ethyl)-N1,N2,N2- tridodecylethane-1,2,-diamine, ethyl(9Z)-17-(dimethylamino)hexacos-9-enoate, (Z)-methyl7- (2-(dimethylamino)-3-(octadec-9-en-1-yloxy)propoxy)heptanoat e, (2Z)-undec-2-en-l-yl8-{[4- (dimethylamino)butanoyl]oxy}heptadecanoate, (14Z)-N,N-dimethylnonacos-14-en-10-amine, ethyl(7Z)-17-(dimethylamino)tricos-7-enoate, (Z)-N1-(2-(4-(2-(Dodec-6-en-l- yl(dodecyl)amino)ethyl)piperazin-N!^^-tridodecylethane-1,2-d iamine, (Z)-methyl5-(2- (dimethylamino)-3-(octadec-9-en-1-yloxy)propoxy)pentanoate, (2- hexylcyclopropyl)methyl10-{[4-(dimethylamino)butanoyl]oxy}no nadecanoate, (15Z)-N,N- dimethylheptacos-15-en-10-amine, ethyl(7Z)-17-(dimethylamino)tetracos-7-enoate, (Z)- methyl4-(2-(dimethylamino)-3-(octadec-9-en-1-yloxy)propoxy)b utanoate, (2Z)-non-2-en-l- yl10-{[4-(dimethylamino)butanoyl]oxy}nonadecanoate, (20Z)-N,N-dimethylheptacos-20-en- 10-amine, N1-(2-(4-(2-(Dioctylamino)ethyl)piperazin-l-yl)ethyl)-N1,N2^ V2- tridodecylethane-l,2-diamine, methyl6-(2-(dimethylamino)-3-((8-(2- octylcyclopropyl)octyl)oxy)propoxy)hexanoate, ethyl6-[2-(9-{[4- (dimethylamino)butanoyl]oxy}octadecyl)cyclopropyl]hexanoate, ethyl(7Z)-17- (dimethylamino)pentacos-7-enoate, 1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine, ethyl(7Z)-17-(dimethylamino)hexacos-7-enoate, (20Z,23Z)-N-ethyl-N-methylnonacosa- 20,23-dien-10-amine, N,N-dimethylheptacosan-10-amine, methyl6-{2-[ll- (dimethylamino)icosyl]cyclopropyl}hexanoate, methyl6-[2-(ll-{[4- (dimethylamino)butanoyl]oxy}icosyl)cyclopropyl]hexanoate, (2-octylcyclopropyl)methyl6- (3-(decyloxy)-2-(dimethylamino)propoxy)hexanoate, methyl8-{2-[9- (dimethylamino)octadecyl]cyclopropyl}octanoate, methyl8-[2-(9-{[4- (dimethylamino)butanoyl]oxy}octadecyl)cyclopropyl]octanoate, methyl7-(2-(8-(2- (dimethylamino)-3-(octyloxy)propoxy)octyl)cyclopropyl)heptan oate, Heptadecan-9-yl8-((2- hydroxyethyl)(tetradecyl)amino)octanoateRepresentative, 2-((2- (Didodecylamino)ethyl)(dodecyl)amino)-l-(4-(2-(didodecylamin o)ethyl)piperazin-1-yl)ethan- 1-one, (2S)-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]undecan-2-amine, (17Z,20Z)-N,N- dimemylhexacosa-17,20-dien-9-amine, (18Z)-heptacos-18-en-10-yl4- (dimethylamino)butanoate, (2S)-1-({6-[3B))-cholest-5-en-3-yloxy]hexyl}oxy)-N,N-dimethy l- 3-[(9Z)-octadec-9-en-1-yloxy]propan-2-amine, methyl10-{2-[7- (dimethylamino)hexadecyl]cyclopropyl}decanoate, methyl10-[2-(7-{[4- (dimethylamino)butanoyl]oxy}hexadecyl)cyclopropyl]decanoate, (2S)-N,N-dimethyl-1-({8- [(lR,2R)-2-{[(lS,2S)-2-pentylcyclopropyl]methyl}cyclopropyl] octyl}oxy)tridecan-2-amine, (2-octylcyclopropyl)methyl6-(2-(dimethylamino)-3-(nonyloxy)p ropoxy)hexanoate, (19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-7-amine, 4-((N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)oxy)pentan-2-yldinonylglycinate, 3-Hydroxybutan-2-ylN-(2- (dinonylamino)ethyl)-N-nonyl, Di(heptadecan-9-yl)8,8'-(26,28-dimethyl-ll,24,30,43-tetraoxo - 10,25,29,44-tetraoxa-19,35-diazatripentacontane-19,35-diyl)d ioctanoate, Di(heptadecan-9- yl)8,8'-(26,27-dimethyl-ll,24,29,42-tetraoxo-10,25,28,43-tet raoxa-19,34- diazadopentacontane-19,34-diyl)dioctanoate, Di(heptadecan-9-yl)8,8'-(ll,24,29,42-tetraoxo- 10,25,28,43-tetraoxa-19,34-diazadopentacontane-19,34-diyl)di octanoate, Di(heptadecan-9- yl)8,8'-((piperazine-l,4-diylbis(5-oxopentane-5,l-diyl))bis( (8-(nonyloxy)-8- oxooctyl)azanediyl))dioctanoate, Di(heptadecan-9-yl)15,18-dimethyl-9,24-bis(8-(nonyloxy)- 8-oxooctyl)-14,19-dioxo-9,15,18,24-tetraazadotriacontanedioa te, Di(heptadecan-9-yl)15,19- dimethyl-9,25-bis(8-(nonyloxy)-8-oxooctyl)-14,20-dioxo-9,15, 19,25- tetraazatritriacontanedioate, Di(heptadecan-9-yl)15,18-diethyl-9,24-bis(8-(nonyloxy)-8- oxooctyl)-14,19-dioxo-9,15,18,24-tetraazadotriacontanedioate , N,N-dimethyl-3-{[(9Z,12Z)- octadeca-9,12-dien-1-yloxy]methyl}dodecan-1-amine, methyl8-[2-(ll-{[4- (dimethylamino)butanoyl]oxy}octadecyl)cyclopropyl]octanoate, methyl8-{2-[ll- (dimethylamino)heptadecyl]cyclopropyl}octanoate(Compound18); , Heptadecan-9-yl8-((2- hydroxyethyl)(8-(nonyloxy)-8-oxooctyl)amino)octanoate, (2-octylcyclopropyl)methyl6-(2- (dimethylamino)-3-(heptyloxy)propoxy)hexanoate, (17Z)-N,N-dimethylhexacos-17-en-9- amine, N1-(2-(4-(2-(Didodecylamino)ethyl)piperazin-l-yl)ethyl)-N1^V 2,N2-trihexylethane- 1,2-diamine, N,N-dimethyl-2-{[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]methyl} undecan-1- amine, methyl8-{2-[ll-(dimethylamino)octadecyl]cyclopropyl}octanoat e, (2- octylcyclopropyl)methyl6-(2-(dimethylamino)-3-(hexyloxy)prop oxy)hexanoate, (18Z)-N,N- dimethylheptacos-18-en-10-amine, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)ethyltetradecanoate, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)ethylnonanoate, TetradecylN-(2-(dinonylamino)ethyl)-N- nonylglycinate, NonylN-(2-(dinonylamino)ethyl)-N-nonylglycinate, 4-(2-((2- (dinonylamino)ethyl)(nonyl)amino)acetamido)butylpentanoate, l,l'-(Piperazine-l,4-diyl)bis(5- (didecylamino)pentan-l-one, 2-((2-(dinonylamino)ethyl)(nonyl)armno)-N- tetradecylacetamide, N-decyl-2-((2-(dinonylamino)ethyl)(nonyl)amino), N1-(3-(3- (dinonylamino)propoxy)propyl)-N1,N2,N2-trinonylethane-l,2-di amine, N1-(2- (dinonylamino)ethyl)-N\N8,N8-trinonyloctane-l,8-diamine, methyl8-[2-(ll-{[4- (dimethylamino)butanoyl]oxy}nonadecyl)cyclopropyl]octanoate, methyl8-{2-[ll- (dimethylamino)nonadecyl]cyclopropyl}octanoate, (Z)-undec-2-en-l-yl6-(3-(decyloxy)-2- (dimethylamino)propoxy)hexanoate, (2R,12Z,15Z)-N,N-dimethyl-1-(undecyloxy)henicosa- 12,15-dien-2-amine, (21Z,24Z)-N,N-dimethyltriaconta-21,24-dien-9-amine, 2- (dinonylamino)-N-(4-(2-((2-(dinonylamino)ethyl)(nonyl)amino) -N-methylacetamido)butyl)- N-methylacetamide, 7,10-dimethyl-13,16-dinonyl-6,ll-dioxo-4-tetradecyl-4,7,10,1 3,16- pentaazapentacosyldecanoate, 2-(dinonylamino)-N-(2-(2-((2- (dinonylamino)ethyl)(nonyl)amino)-N-ethylacetamido)ethyl)-N- ethylacetamide, 2- (dinonylamino)-N-(3-(2-((2-(dinonylamino)ethyl)(nonyl)amino) -N- methylacetamido)propyl)-N-methylacetamide, 2-((2-(di((Z)-non-3-en-l-yl)amino)ethyl)((Z)- non-3-en-l-yl)amino)-N-(2-(2-(dinonylamino)-N-methylacetamid o)ethyl)-N- methylacetamide, 2-(dinonylamino)-N-(2-(2-((2- (dinonylamino)ethyl)(nonyl)amino)acetamido)ethyl)acetamide, Pentyl8,1l-dimethyl-5,14,17- trinonyl-7,12-dioxo-5,8,l1,14,17-pentaazahexacosanoate2-((2- (Dinonylamino)ethyl)(nonyl)aniino)-N-methyl-N-(2-(methylandn o)ethyl)acetami, 2- (Dinonylamino)-N-(2-(2-((2-(dinonylamino)ethyl)(nonyl)amino) -N-methylacetamido)ethyl)- N-methylacetamide2-(Dinonylamino)-N-methyl-N-(2-(methylamino )ethyl)acetamide, 2-((N- (2-(Dinonylamino)ethyl)-N-nonylglycyl)oxy)ethyldinonylglycin ate2- Hydroxyethyldinonylglycinate, methyl8-[2-(ll-{[4- (dimethylamino)butanoyl]oxy}icosyl)cyclopropyl]octanoate, methyl8-{2-[ll- (dimethylamino)icosyl]cyclopropyl}octanoate, (Z)-undec-2-en-l-yl6-(2-(dimethylamino)-3- (nonyloxy)propoxy)hexanoate, (2R,12Z,15Z)-1-(hexadecyloxy)-N,N-dimethylhenicosa- 12,15-dien-2-amine, (22Z,25Z)-N,N-dimethylhentriaconta-22,25-dien-10-amine, l,l- (Piperazine-l,4-diyl)bis(4-(didecylamino)butan-l-one)fert-Bu tyl4-(didecylaminobutanoate, Heptyl5-(4-(N-(2-(dinonylamino)ethyl)-N-nonylglycyl)piperazi n-l-yl)-5-oxopentanoate5- (Heptloxy)-5-oxopentanoicacid, Heptyl5-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-5-oxopentanoate5-(Heptloxy)-5-ox opentanoic, (Z)-4-((2-(4-(N- (2-(dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoe thyl)(tetradecyl)amino)but-2- en-1-y1nonanoate(Z)-4-Hydroxybut-2-en-l-ylnonanoate, (Z)-3-((2-(4-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoethy l)(tetradec-9-en-1- yl)amino)propy1decanoate(Z)-Tetradec-9-en-l-ylmethanesulfona te, methyl8-[2-(9-{[4- (dimethylamino)butanoyl]oxy}pentadecyl)cyclopropyl]octanoate , methyl8-{2-[9- (dimethylamino)pentadecyl]cyclopropyl}octanoate, (Z)-undec-2-en-l-yl6-(2- (dimethylamino)-3-(heptyloxy)propoxy)hexanoate, (2R,12Z,15Z)-1-(hexyloxy)-N,N- dimethylhenicosa-12,15-dien-2-amine, (16Z,19Z)-N,N-dimethylpentacosa-16,19-dien-6- amine, Methyl8-((2-(4-(N-(2-(Di((Z)-non-3-en-l-yl)amino)ethyl)-N-(( Z)-non-3-en-l- yl)glycyl)piperazin-l-yl)-2-oxoethyl)(nonyl)amino)octanoatef ert-Butyl4- (nonylglycyl)piperazine-1-carboxylate, 3-((2-(4-(N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(tetradecyl)amino)pro pyl(Z)-dec-3-enoate(Z)-Dec-3- en-l-ol, 2-((2-(Di((Z)-non-3-en-l-yl)amino)ethyl)((Z)-non-3-en-l-yl)a mino)-l-(4- (dinonylglycyl)piperazin-1-yl)ethan-1-one(Z)-1-Bromonon-4-en e, 3-((2-(4-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperazin- oxoethyl)(dodecyl)amino)propyloctanoatetot-Butyldodecylglyci nate, S-Pentyl4-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoethy l)(nonyl)amino)butanethioate, 3-((2-(l-(N-(2-(Dinonylamino)ethyl)-N-nonylglycyl)piperidin- ^yl)ethyl)(nonyl)amino)propyl3-methylhexanoatefert-Butyl4-(2 -((3-((3- methylhexanoyl)oxy)propyl)(nonyl)amino)ethyl)piperidine-l-, 3-((2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(non yl)amino)-2- methylpropylhexanoate, 3-((2-(4-(N-(2-(Dinonylamino)ethyl)-N-nonylglycyl)piperazin- oxoethyl)(nonyl)amino)propyl3-methylhexanoate, 3-((2-(4-(N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)piperazin-oxoethyl)(nonyl)amino)-2-methylpropylh exanoate, methyl8-[2-(9-{[4- (dimethylamino)butanoyl]oxy}hexadecyl)cyclopropyl]octanoate, methyl8-{2-[9- (dimethylamino)hexadecyl]cyclopropyl}octanoate, (Z)-undec-2-en-l-yl6-(2-(dimethylamino)- 3-(hexyloxy)propoxy)hexanoate, (2R,12Z,15Z)-1-(decyloxy)-N,N-dimethylhenicosa-l2,15- dien-2-amine, (17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-7-amine, 2-((2- (Dinonylamino)ethyl)(nonyl)amino)ethyl1-(dinonylglycyl)piper idine-4-carboxylate, l-(2- (Dinonylamino)ethyl)4-(2-((2-(dinonylamino)ethyl)(nonyl)amin o)ethyl)cyclohexane-1,4- dicarboxylate2-(Dinonylamino)ethan-1-ol, Methyl12-((2-(l-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)pyrrolidin-3-yl)ethyl)(tetradecyl)amino)dodecano atefert-Butyl3-(2-((12- methoxy-12-oxododecyl)(tetradecyl)amino)ethyl)pyrrolidine-l- carboxylate, 3-((2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3- yl)ethyl)(tetradecyl)amino)propyldecanoateter/-Butyl3-(2-((3 - (decanoyloxy)propyl)(tetradecyl)amino)ethyl)pyrrolidine-l-ca rboxylate, "Heptyl6-((2-(l-(N- (2-(dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3- yl)ethyl)(tetradecyl)amino)hexanoatetot-Butyl3-(2-((6-(hepty loxy)-6- oxohexyl)(tetradecyl)amino)ethyl)pyrrolidine-1-carboxylate, ", Pentyl8-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3-yl)ethyl)(te tradecyl)amino)octanoate/er/- Butyl3-(2-(tetradecylamino)ethyl)pyrrolidine-l-carboxylate, Methyl12-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-3-yl)ethyl)(tet radecyl)amino)dodecanoate- Butyl3-(2-((12-methoxy-12-oxododecyl)(tetradecyl)amino)ethyl )piperidine-l-carboxylate, 3- ((2-(l-(N-(2-(Dinonylamino)ethyl)-N-nonylglycyl)piperidin-3- yl)ethyl)(tetradecyl)amino)propyldecanoate, Heptyl6-((2-(l-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperidin-3-yl)ethyl)(tetradecyl)amino)hexanoate , Pentyl8-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-3-yl)ethyl)(tet radecyl)amino)octanoate, Pentyl6-((2-(4-(2-((2-(didodecylamino)ethyl)(dodecyl)amino)e thyl)piperazin-l- yl)ethyl)(dodecyl)amino)hexanoateStep1:Pentyl6-bromohexanoat e, methyl8-[2-(9-{[4- (dimethylamino)butanoyl]oxy}heptadecyl)cyclopropyl]octanoate , methyl8-{2-[9- (dimethylamino)heptadecyl]cyclopropyl}octanoate, (2S,12Z,15Z)-N,N-dimethyl-1- (octyloxy)henicosa-12,15-dien-2-amine, (2-octylcyclopropyl)methyl6-(2-(dimethylamino)-3- (octyloxy)propoxy)hexanoate, (18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-8-amine, trans- 1-methyl-3,4-bis(((Z)-hexadec-9-enoyloxy)methyl)pyrrolidine, (Z)-Non-2-en-l-yl4-((2-(4-(N- (2-(dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2- oxoethyl)(tetradecyl)amino)butanoate, trans-1-methyl-3,4-bis(((9Z,12Z)-octadeca-9,12- dienoyloxy)methyl)pyrrolidine, Methyl12-((2-(4-(N-(2-(dinonylamino)ethyl)-N- nonylglycyl)piperazin-l-yl)-2-oxoethyl)(tetradecyl)amino)dod ecanoate, ethyl(7Z)-17-[2- (dimethylamino)ethyl]hexacos-7-enoate, trans-1-methyl-3,4-bis(((Z)-octadeca-9- enoyloxy)methyl)pyrrolidine, methyl6-(2-{]11-^2- (dimethylamino)ethyl]icosyl}cyclopropyl)hexanoate, Methyl12-((2-(l-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyl)(tet radecyl)amino)dodecanoate, methyl10-(2-V-^2-(dimethylamino)ethyl]hexadecyl}cyclopropyl) decanoate, methyl8-(2- {111-;2-(dimethylamino)ethyl]heptadecyl}cyclopropyl)octanoat e, 2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperidin-4-yl)ethyldinon ylglycinatefert-Butyl4-(2- ((dinonylglycyl)oxy)ethyl)piperidine-1-carboxylate, methyl8-(2-{lLl-;2- (dimethylamino)ethyl]octadecyl}cyclopropyl)octanoate, methyl8-(2-{l11- "2- (dimethylamino)ethyl]nonadecyl}cyclopropyl)octanoate, l,-(piperazine-l,4-diyl)bis(2- (dinonylamino)ethan-l-one), methyl8-[2-{]11-^2- (dimethylamino)ethyl]icosyl}cyclopropyl)octanoate, methyl8-(2-{9-[2- (dimethylamino)ethyl]pentadecyl}cyclopropyl)octanoate, methyl(7Z)-19-{[4- (dimethylamino)butanoyl]oxy}octacos-7-enoate, methyl(7Z)-19-(dimethylamino)octacos-7- enoate, cis-1-methyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-4-(octy loxy)pyrrolidine, 2- (Didodecylamino)-l-(4-(N-(2-(didodecylamino)ethyl)-N-dodecyl glycyl)piperazin-l-yl)ethan- 1-one, (Z)-undec-2-en-l-yl6-(2-(dimethylamino)-3-(octyloxy)propoxy) hexanoate, (2SN,N- dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]decan-2-amin e(Compound11), (19Z,22Z)- N,N-dimeihyloctacosa-19,22-dien-9-amine, methyl8-(2-{9-[2- (dimethylamino)ethyl]hexadecyl}cyclopropyl)octanoate, 5-((2-(4-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)piperazin- oxoethyl)(nonyl)amino)pentylmethylcarbonate, methyl8-(2-{9-[2- (dimethylamino)ethyl]heptadecyl}cyclopropyl)octanoate, methyl(7Z)-19-[2- (dimethylamino)ethyl]octacos-7-enoate, (Z)-Pent-2-en-l-yl4-((2-(4-(N-(2- (dinonylamino)ethyl)-N-nonylglycyl)piperazin-l-yl)-2-oxoethy l)(nonyl)amino)butanoate, methyl(1lZ)-19-[2-(dimethylamino)ethyl]octacos-l1-enoate, methyl(9Z)-21-[2- (dimethylamino)ethyl]heptacos-9-enoate, methyl(9Z)-21-[2-(dimethylamino)ethyl]octacos-9- enoate, methyl(9Z)-21-[2-(dimethylamino)ethyl]nonacos-9-enoate, 2-(l-(N-(2- (Dinonylamino)ethyl)-N-nonylglycyl)pyrrolidin-3-yl)ethyldino nylglycinate, methyl(9Z)-21- [2-(dimethylamino)ethyl]triacont-9-enoate, (l-(N-(2-(Dinonylamino)ethyl)-N- nonylglycyl)pyrrolidin-3-yl)methyldinonylglycinate, methyl(9Z)-19-[2- (dimethylamino)ethyl]pentacos-9-enoate, methyl(9Z)-19-[2-(dimethylamino)ethyl]hexacos-9- enoate, methyl6-(2-(8-(3-(decyloxy)-2- (dimethylamino)propoxy)octyl)cyclopropyl)hexanoate, methyl(1lZ)-19-{[4- (dimethylamino)butanoyl]oxy}octacos-l1-enoate, methyl(1lZ)-19-(dimethylamino)octacos- l1-enoate, (2S)-N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]dod ecan-2-amine, (14Z,17Z)-N,N-dimethyltricosa-14,17-dien-4-amine, Methyldi((9Z,12Z)-octadeca-9,12- dienyl)amine, methyl(9Z)-19-{[4-(dimethylamino)butanoyl]oxy}octacos-9-enoa te, methyl(9Z)-19-(dimethylamino)octacos-9-enoate, (Z)-methyl17-(2-(dimethylamino)-3- (octyloxy)propoxy)heptadec-8-enoate, (3R,4R)-3,4-bis((Z)-hexadec-9-enyloxy)-1- methylpyrrolidine, (2S)-N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]und ecan-2- amine, (20Z,23Z)-nonacosa-20,23-dien-10-yl4-(dimethylamino)butanoat e, (20Z,23Z)-N,N- dimethylnonacosa-20,23-dien-10-amine, 3-((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31- tetraen-19-yloxy)-N,N-dimethylpropan-l-amine, 3-((6Z,9Z,28Z,31Z)-heptatriaconta- 6,9,28,31-tetraen-19-yloxy)-N,N-dimethylpropan-1-amine, (6Z,9Z,28Z,31Z)-heptatriaconta- 6,9,28,31-tetraen-19-yl4-(dimethylamino)butanoate), (6Z,16Z)-12-((Z)-dec-4-enyl)docosa- 6,16-dien-l1-yl5-(dimethylamino)pentanoate, (6Z,16Z)-12-((Z)-dec-4-enyl)docosa-6,16-dien- l1-yl5-(dimethylamino)pentanoat, (6Z,16Z)-12-((Z)-dec-4-enyl)docosa-6,16-dien-11-yl5- (dimethylamino)pentanoate, L-arginine-alpha-(2,3-dilauryloxy)propylamide, L-lysine-alpha- (2,3-dilauryloxy)propylamide, 2,3-dioleyloxypropylamine, 2,3-distearyloxypropylamine, 2,3- dilauryloxypropylamine, dilinoleylmethyl4-(dimethylamino)propylether), dilinoleylmethyl4- (dimethylamino)butylether), and 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[l,3]-dioxolane. [0637] In some embodiments, the at least one non-cationic lipid comprises at least one phospholipid, at least one fusogenic lipid, at least one anionic lipid, at least one helper lipid, at least one neutral lipid, or any combination thereof. In some embodiments, the LNP may be essentially devoid of the at least one non-cationic lipid. In some embodiments, the LNP may contain no amount of the at least one non-cationic lipid. [0638] In some embodiments, at least one non-cationic lipid may be selected from, but is not limited to, at least one of 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), DSPC but with 3 unsaturated double bonds pertail (18:3 PC), Acylcarnosine (AC), 1- hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), N-oleoyl-SPM (C18:l), N- lignocerylSPM (C24:0), N-nervacylC (C24:l), carbamoyl]cholesterol (Cet-P), cholesterolhemisuccinate (CHEMS), cholesterol (Chol), Cholesterolhemidodecanedicarboxylic acid (Chol-C12), 12- Cholesteryloxycarbonylaminododecanoic acid (Chol-C13N), Cholesterolhemioxalate (Chol- C2), Cholesterolhemimalonate (Chol-C3), N-(Cholesteryl-oxycarbonyl)glycine (Chol-C3N), Cholesterolhemiglutarate (Chol-C5), Cholesterolhemiadipate (Chol-C6), Cholesterolhemipimelate (Chol-C7), Cholesterolhemisuberate (Chol-C8), Cardiolipid (CL), l,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC8-9PC), dicetylphosphate (DCP), dihexadecylphosphate (DCP1), 1,2-Dipalmitoyglycerol-3-hemisuccinate (DGSucc), short-chainbis-n-heptadecanoylphosphatidylcholine (DHPC), dihexadecoylphosphoethanolamine (DHPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), l,2-dilauroyl-sn-glycero-3-PE (DLPE), Dimyristoylglycerolhemisuccinate (DMGS), dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleyloxybenzylalcohol (DOBA), 1,2- dioleoylglyceryl-3-hemisuccinate (DOGHEMS), N-[2~(2-{2-[2-(2,3-Bis-octadec-9-enyloxy- propoxy)-ethoxy]-ethoxy}-ethoxy)-ethyl]-3-(3,4,5-1rihydroxy- 6-hydroxymethyl-1etrahydro- pyran-2-ylsulfanyl)-propionamide (DOGP4αMan), dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylethanolamine (DOPE), dioleoyl-phosphatidylethanolamine4-(N- maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell-fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl-phosphatidyl-ethanolamine (DSPE), distearoylphosphoethanolamineimidazole (DSPEI), 1,2-diundecanoyl-sn-glycero- phosphocholine (DUPC), eggphosphatidylcholine (EPC), N-histidinylcholesterolcarbamate (HCChol), histaminedistearoylglycerol (HDSG), N-histidinylcholesterolhemisuccinate (HistChol), 1,2-Dipalmitoylglycerol-hemisuccinate-Nα-Histidinyl-Hemisuc cinate (HistSuccDG), N-(5'-hydroxy-3'-oxypentyl)-10-12-pentacosadiynamide (h-Pegi-PCDA), 2-[l- hexyloxyethyl]-2-devinylpyropheophorbide-a (HPPH), hydrogenatedsoybeanphosphatidylcholine (HSPC), 1,2-Dipalmitoylglycerol-Oα-histidinyl- Nα-hemisuccinate (IsohistsuccDG), mannosialized dipalmitoylphosphatidylethanolamine (ManDOG), l,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine-N-[4-(p- maleimidomethyl)cyclohexane-carboxamide] (MCC-PE), 1,2-diphytanoyl-sn-glycero-3- phosphoethanolamine (ME 16.0 PE), 1-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC), a thiol-reactive maleimide head group lipid, e.g., 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine-N-[4-(p-maleimidophenyl)but-yramid (MPB-PE), Nervonic Acid (NA), sodiumcholate (NaChol), l,2-dioleoyl-sn-glycero-3-[phosphoethanolamine-N-dodecanoyl (NC12-DOPE), defined by synthesis example in WO2008042973A2 (ND98), "N- glutarylphosphatidylethanolamine(s) of Formula 1" (NG-PE), N-hydroxysulfosuccinimide (NHS-'x'), "N~(co)-dicarboxylicacid-derivatized phosphatidylethanolamines encompassed by Formula 1" (NωPE-'x'), OleicAcid (OA), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3- phosphocholine (OChemsPC), phosphatidicacid (PA), phosphatidylethanolamine lipid (PE), PE lipid conjugated with polyethyleneglycol (PEG). One example of PEG-PE can be polyethyleneglycol-distearoylphosphatidylethanolamine lipid (PEG-PE), phosphatidylglycerol (PG), partially hydrogenated soy phosphatidylchloline (PHSPC), phosphatidylinositol lipid (PI), phosphotidylinositol-4-phosphate (PIP), palmitoyloleoylphosphatidylcholine (POPC), phosphatidylethanolamine (POPE), palmitoyloleyolphosphatidylglycerol (POPG), phosphatidylserine (PS), lissaminerhodamineB-phosphatidylethanolamine lipid (Rh-PE), purifiedsoy-derived mixture of phospholipids (SIOO), phosphatidylcholine (SM), 18-1-transPE,1-stearoyl-2-oleoyl- phosphatidyethanolamine (SOPE), soybeanphosphatidylcholine (SPC), sphingomyelins (SPM), alpha.alpha'-trehalose6,6'-dibehenate (TDB), l,2-dielaidoyl-sn-glycero-3- phophoethanolamine (transDOPE), ((23S,5R)-3-(bis(hexadecyloxy)methoxy)-5-(5-methyl- 2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran-2-yl )methylmethylphosphate, 1,2- diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3- phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2- didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3- phosphocholine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn- glycero-3-phosphoethanolamine, 1,2-dioleyl-sn-glycero-3-phosphoethanolamine, 1,2- distearoyl-sn-glycero-3-phosphoethanolamine, 16-O-monomethyl PE, 16-O-dimethyl PE, and dioleylphosphatidylethanolamine. [0639] In some embodiments, the LNP comprises an ionizable lipid or lipid-like material. As a non-limiting example, the ionizable lipid may be C12-200, CKK-E12, 5A2-SC8, BAMEA- 016B, or 7C1. Other ionizable lipids are known in the art and are useful herein. [0640] In some embodiments, the LNP comprises a phospholipid. As a non-limiting example, the phospholipid (helper) may be DOPE, DSPC, DOTAP, or DOTMA. [0641] In some embodiments, the LNP comprises a PEG derivative. As a non-limiting example, the PEG derivative may be a lipid-anchored such as PEG is C14-PEG2000, C14- PEG1000, C14-PEG3000, C14-PEG5000, C12-PEG1000, C12-PEG2000, C12-PEG3000, C12-PEG5000, C16-PEG1000, C16-PEG2000, C16-PEG3000, C16-PEG5000, C18- PEG1000, C18-PEG2000, C18-PEG3000, or C18-PEG5000. [0642] In some embodiments, the at least one sterol comprises at least one cholesterol or cholesterol derivative. In some embodiments, the LNP may be essentially devoid of an at least one sterol. In some embodiments, the LNP may contain no amount of the at least one sterol. [0643] In some embodiments, the at least one particle-activity-modifying-agent comprises at least one component that reduced aggregation of particles, at least one component that decreases clearing of the LNP from circulation in a subject, at least component that increases the LNP's ability to traverse mucus layers, at least one component that decreases a subjects immune response to administration of the LNP, at least one component that modifies membrane fluidity of the LNP, at least one component that contributes to the stability of the LNP, or any combination thereof. In some embodiments, the LNP may be essentially devoid of the at least one particle-activity-modifying-agent. In some embodiments, the LNP may contain no amount of the at least one particle-activity-modifying-agent. [0644] In some embodiments, the particle-activity-modifying-agent may be comprised of a polymer. In some embodiments, the polymer comprising the particle-activity-modifying-agent may be comprised of at least one polyethylene glycol (PEG), at least one polypropylene glycol (PPG), poly(2-oxazoline) (POZ), at least one polyamide (ATTA), at least one cationic polymer, or any combination thereof. [0645] In some embodiments, the average molecular weight of the polymer moiety (e.g., PEG) may be between 500 and 20,000 daltons. In some embodiments, the molecular weight of the polymer may be about 500 to 20,000, 1,000 to 20,000, 1,500 to 20,000, 2,000 to 20,000, 2,500 to 20,000, 3,000 to 20,000, 3,500 to 20,000, 4,000 to 20,000, 4,500 to 20,000, 5,000 to 20,000, 5,500 to 20,000, 6,000 to 20,000, 6,500 to 20,000, 7,000 to 20,000, 7,500 to 20,000, 8,000 to 20,000, 8,500 to 20,000, 9,000 to 20,000, 9,500 to 20,000, 10,000 to 20,000, 10,500 to 20,000, 11,000 to 20,000, 11,500 to 20,000, 12,000 to 20,000, 12,500 to 20,000, 13,000 to 20,000, 13,500 to 20,000, 14,000 to 20,000, 14,500 to 20,000, 15,000 to 20,000, 15,500 to 20,000, 16,000 to 20,000, 16,500 to 20,000, 17,000 to 20,000, 17,500 to 20,000, 18,000 to 20,000, 18,500 to 20,000, 19,000 to 20,000, 19,500 to 20,000, 500 to 19,500, 1,000 to 19,500, 1,500 to 19,500, 2,000 to 19,500, 2,500 to 19,500, 3,000 to 19,500, 3,500 to 19,500, 4,000 to 19,500, 4,500 to 19,500, 5,000 to 19,500, 5,500 to 19,500, 6,000 to 19,500, 6,500 to 19,500, 7,000 to 19,500, 7,500 to 19,500, 8,000 to 19,500, 8,500 to 19,500, 9,000 to 19,500, 9,500 to 19,500, 10,000 to 19,500, 10,500 to 19,500, 11,000 to 19,500, 11,500 to 19,500, 12,000 to 19,500, 12,500 to 19,500, 13,000 to 19,500, 13,500 to 19,500, 14,000 to 19,500, 14,500 to 19,500, 15,000 to 19,500, 15,500 to 19,500, 16,000 to 19,500, 16,500 to 19,500, 17,000 to 19,500, 17,500 to 19,500, 18,000 to 19,500, 18,500 to 19,500, 19,000 to 19,500, 1,500 to 19,000, 2,000 to 19,000, 2,500 to 19,000, 3,000 to 19,000, 3,500 to 19,000, 4,000 to 19,000, 4,500 to 19,000, 5,000 to 19,000, 5,500 to 19,000, 6,000 to 19,000, 6,500 to 19,000, 7,000 to 19,000, 7,500 to 19,000, 8,000 to 19,000, 8,500 to 19,000, 9,000 to 19,000, 9,500 to 19,000, 10,000 to 19,000, 10,500 to 19,000, 11,000 to 19,000, 11,500 to 19,000, 12,000 to 19,000, 12,500 to 19,000, 13,000 to 19,000, 13,500 to 19,000, 14,000 to 19,000, 14,500 to 19,000, 15,000 to 19,000, 15,500 to 19,000, 16,000 to 19,000, 16,500 to 19,000, 17,000 to 19,000, 17,500 to 19,000, 18,000 to 19,000, 18,500 to 19,000, 1,500 to 18,500, 2,000 to 18,500, 2,500 to 18,500, 3,000 to 18,500, 3,500 to 18,500, 4,000 to 18,500, 4,500 to 18,500, 5,000 to 18,500, 5,500 to 18,500, 6,000 to 18,500, 6,500 to 18,500, 7,000 to 18,500, 7,500 to 18,500, 8,000 to 18,500, 8,500 to 18,500, 9,000 to 18,500, 9,500 to 18,500, 10,000 to 18,500, 10,500 to 18,500, 11,000 to 18,500, 11,500 to 18,500, 12,000 to 18,500, 12,500 to 18,500, 13,000 to 18,500, 13,500 to 18,500, 14,000 to 18,500, 14,500 to 18,500, 15,000 to 18,500, 15,500 to 18,500, 16,000 to 18,500, 16,500 to 18,500, 17,000 to 18,500, 17,500 to 18,500, 18,000 to 18,500, 1,500 to 18,000, 2,000 to 18,000, 2,500 to 18,000, 3,000 to 18,000, 3,500 to 18,000, 4,000 to 18,000, 4,500 to 18,000, 5,000 to 18,000, 5,500 to 18,000, 6,000 to 18,000, 6,500 to 18,000, 7,000 to 18,000, 7,500 to 18,000, 8,000 to 18,000, 8,500 to 18,000, 9,000 to 18,000, 9,500 to 18,000, 10,000 to 18,000, 10,500 to 18,000, 11,000 to 18,000, 11,500 to 18,000, 12,000 to 18,000, 12,500 to 18,000, 13,000 to 18,000, 13,500 to 18,000, 14,000 to 18,000, 14,500 to 18,000, 15,000 to 18,000, 15,500 to 18,000, 16,000 to 18,000, 16,500 to 18,000, 17,000 to 18,000, 17,500 to 18,000, 1,500 to 17,500, 2,000 to 17,500, 2,500 to 17,500, 3,000 to 17,500, 3,500 to 17,500, 4,000 to 17,500, 4,500 to 17,500, 5,000 to 17,500, 5,500 to 17,500, 6,000 to 17,500, 6,500 to 17,500, 7,000 to 17,500, 7,500 to 17,500, 8,000 to 17,500, 8,500 to 17,500, 9,000 to 17,500, 9,500 to 17,500, 10,000 to 17,500, 10,500 to 17,500, 11,000 to 17,500, 11,500 to 17,500, 12,000 to 17,500, 12,500 to 17,500, 13,000 to 17,500, 13,500 to 17,500, 14,000 to 17,500, 14,500 to 17,500, 15,000 to 17,500, 15,500 to 17,500, 16,000 to 17,500, 16,500 to 17,500, 17,000 to 17,500, 1,500 to 17,000, 2,000 to 17,000, 2,500 to 17,000, 3,000 to 17,000, 3,500 to 17,000, 4,000 to 17,000, 4,500 to 17,000, 5,000 to 17,000, 5,500 to 17,000, 6,000 to 17,000, 6,500 to 17,000, 7,000 to 17,000, 7,500 to 17,000, 8,000 to 17,000, 8,500 to 17,000, 9,000 to 17,000, 9,500 to 17,000, 10,000 to 17,000, 10,500 to 17,000, 11,000 to 17,000, 11,500 to 17,000, 12,000 to 17,000, 12,500 to 17,000, 13,000 to 17,000, 13,500 to 17,000, 14,000 to 17,000, 14,500 to 17,000, 15,000 to 17,000, 15,500 to 17,000, 16,000 to 17,000, 16,500 to 17,000, 1,500 to 16,500, 2,000 to 16,500, 2,500 to 16,500, 3,000 to 16,500, 3,500 to 16,500, 4,000 to 16,500, 4,500 to 16,500, 5,000 to 16,500, 5,500 to 16,500, 6,000 to 16,500, 6,500 to 16,500, 7,000 to 16,500, 7,500 to 16,500, 8,000 to 16,500, 8,500 to 16,500, 9,000 to 16,500, 9,500 to 16,500, 10,000 to 16,500, 10,500 to 16,500, 11,000 to 16,500, 11,500 to 16,500, 12,000 to 16,500, 12,500 to 16,500, 13,000 to 16,500, 13,500 to 16,500, 14,000 to 16,500, 14,500 to 16,500, 15,000 to 16,500, 15,500 to 16,500, 16,000 to 16,500, 1,500 to 16,000, 2,000 to 16,000, 2,500 to 16,000, 3,000 to 16,000, 3,500 to 16,000, 4,000 to 16,000, 4,500 to 16,000, 5,000 to 16,000, 5,500 to 16,000, 6,000 to 16,000, 6,500 to 16,000, 7,000 to 16,000, 7,500 to 16,000, 8,000 to 16,000, 8,500 to 16,000, 9,000 to 16,000, 9,500 to 16,000, 10,000 to 16,000, 10,500 to 16,000, 11,000 to 16,000, 11,500 to 16,000, 12,000 to 16,000, 12,500 to 16,000, 13,000 to 16,000, 13,500 to 16,000, 14,000 to 16,000, 14,500 to 16,000, 15,000 to 16,000, 15,500 to 16,000, 1,500 to 15,500, 2,000 to 15,500, 2,500 to 15,500, 3,000 to 15,500, 3,500 to 15,500, 4,000 to 15,500, 4,500 to 15,500, 5,000 to 15,500, 5,500 to 15,500, 6,000 to 15,500, 6,500 to 15,500, 7,000 to 15,500, 7,500 to 15,500, 8,000 to 15,500, 8,500 to 15,500, 9,000 to 15,500, 9,500 to 15,500, 10,000 to 15,500, 10,500 to 15,500, 11,000 to 15,500, 11,500 to 15,500, 12,000 to 15,500, 12,500 to 15,500, 13,000 to 15,500, 13,500 to 15,500, 14,000 to 15,500, 14,500 to 15,500, 15,000 to 15,500, 1,500 to 15,000, 2,000 to 15,000, 2,500 to 15,000, 3,000 to 15,000, 3,500 to 15,000, 4,000 to 15,000, 4,500 to 15,000, 5,000 to 15,000, 5,500 to 15,000, 6,000 to 15,000, 6,500 to 15,000, 7,000 to 15,000, 7,500 to 15,000, 8,000 to 15,000, 8,500 to 15,000, 9,000 to 15,000, 9,500 to 15,000, 10,000 to 15,000, 10,500 to 15,000, 11,000 to 15,000, 11,500 to 15,000, 12,000 to 15,000, 12,500 to 15,000, 13,000 to 15,000, 13,500 to 15,000, 14,000 to 15,000, 14,500 to 15,000, 1,500 to 14,500, 2,000 to 14,500, 2,500 to 14,500, 3,000 to 14,500, 3,500 to 14,500, 4,000 to 14,500, 4,500 to 14,500, 5,000 to 14,500, 5,500 to 14,500, 6,000 to 14,500, 6,500 to 14,500, 7,000 to 14,500, 7,500 to 14,500, 8,000 to 14,500, 8,500 to 14,500, 9,000 to 14,500, 9,500 to 14,500, 10,000 to 14,500, 10,500 to 14,500, 11,000 to 14,500, 11,500 to 14,500, 12,000 to 14,500, 12,500 to 14,500, 13,000 to 14,500, 13,500 to 14,500, 14,000 to 14,500, 1,500 to 14,000, 2,000 to 14,000, 2,500 to 14,000, 3,000 to 14,000, 3,500 to 14,000, 4,000 to 14,000, 4,500 to 14,000, 5,000 to 14,000, 5,500 to 14,000, 6,000 to 14,000, 6,500 to 14,000, 7,000 to 14,000, 7,500 to 14,000, 8,000 to 14,000, 8,500 to 14,000, 9,000 to 14,000, 9,500 to 14,000, 10,000 to 14,000, 10,500 to 14,000, 11,000 to 14,000, 11,500 to 14,000, 12,000 to 14,000, 12,500 to 14,000, 13,000 to 14,000, 13,500 to 14,000, 1,500 to 13,500, 2,000 to 13,500, 2,500 to 13,500, 3,000 to 13,500, 3,500 to 13,500, 4,000 to 13,500, 4,500 to 13,500, 5,000 to 13,500, 5,500 to 13,500, 6,000 to 13,500, 6,500 to 13,500, 7,000 to 13,500, 7,500 to 13,500, 8,000 to 13,500, 8,500 to 13,500, 9,000 to 13,500, 9,500 to 13,500, 10,000 to 13,500, 10,500 to 13,500, 11,000 to 13,500, 11,500 to 13,500, 12,000 to 13,500, 12,500 to 13,500, 13,000 to 13,500, 1,500 to 13,000, 2,000 to 13,000, 2,500 to 13,000, 3,000 to 13,000, 3,500 to 13,000, 4,000 to 13,000, 4,500 to 13,000, 5,000 to 13,000, 5,500 to 13,000, 6,000 to 13,000, 6,500 to 13,000, 7,000 to 13,000, 7,500 to 13,000, 8,000 to 13,000, 8,500 to 13,000, 9,000 to 13,000, 9,500 to 13,000, 10,000 to 13,000, 10,500 to 13,000, 11,000 to 13,000, 11,500 to 13,000, 12,000 to 13,000, 12,500 to 13,000, 1,500 to 12,500, 2,000 to 12,500, 2,500 to 12,500, 3,000 to 12,500, 3,500 to 12,500, 4,000 to 12,500, 4,500 to 12,500, 5,000 to 12,500, 5,500 to 12,500, 6,000 to 12,500, 6,500 to 12,500, 7,000 to 12,500, 7,500 to 12,500, 8,000 to 12,500, 8,500 to 12,500, 9,000 to 12,500, 9,500 to 12,500, 10,000 to 12,500, 10,500 to 12,500, 11,000 to 12,500, 11,500 to 12,500, 12,000 to 12,500, 1,500 to 12,000, 2,000 to 12,000, 2,500 to 12,000, 3,000 to 12,000, 3,500 to 12,000, 4,000 to 12,000, 4,500 to 12,000, 5,000 to 12,000, 5,500 to 12,000, 6,000 to 12,000, 6,500 to 12,000, 7,000 to 12,000, 7,500 to 12,000, 8,000 to 12,000, 8,500 to 12,000, 9,000 to 12,000, 9,500 to 12,000, 10,000 to 12,000, 10,500 to 12,000, 11,000 to 12,000, 11,500 to 12,000, 1,500 to 11,500, 2,000 to 11,500, 2,500 to 11,500, 3,000 to 11,500, 3,500 to 11,500, 4,000 to 11,500, 4,500 to 11,500, 5,000 to 11,500, 5,500 to 11,500, 6,000 to 11,500, 6,500 to 11,500, 7,000 to 11,500, 7,500 to 11,500, 8,000 to 11,500, 8,500 to 11,500, 9,000 to 11,500, 9,500 to 11,500, 10,000 to 11,500, 10,500 to 11,500, 11,000 to 11,500, 1,500 to 11,000, 2,000 to 11,000, 2,500 to 11,000, 3,000 to 11,000, 3,500 to 11,000, 4,000 to 11,000, 4,500 to 11,000, 5,000 to 11,000, 5,500 to 11,000, 6,000 to 11,000, 6,500 to 11,000, 7,000 to 11,000, 7,500 to 11,000, 8,000 to 11,000, 8,500 to 11,000, 9,000 to 11,000, 9,500 to 11,000, 10,000 to 11,000, 10,500 to 11,000, 1,500 to 10,500, 2,000 to 10,500, 2,500 to 10,500, 3,000 to 10,500, 3,500 to 10,500, 4,000 to 10,500, 4,500 to 10,500, 5,000 to 10,500, 5,500 to 10,500, 6,000 to 10,500, 6,500 to 10,500, 7,000 to 10,500, 7,500 to 10,500, 8,000 to 10,500, 8,500 to 10,500, 9,000 to 10,500, 9,500 to 10,500, 10,000 to 10,500, 1,500 to 10,000, 2,000 to 10,000, 2,500 to 10,000, 3,000 to 10,000, 3,500 to 10,000, 4,000 to 10,000, 4,500 to 10,000, 5,000 to 10,000, 5,500 to 10,000, 6,000 to 10,000, 6,500 to 10,000, 7,000 to 10,000, 7,500 to 10,000, 8,000 to 10,000, 8,500 to 10,000, 9,000 to 10,000, 9,500 to 10,000, 1,500 to 9,500, 2,000 to 9,500, 2,500 to 9,500, 3,000 to 9,500, 3,500 to 9,500, 4,000 to 9,500, 4,500 to 9,500, 5,000 to 9,500, 5,500 to 9,500, 6,000 to 9,500, 6,500 to 9,500, 7,000 to 9,500, 7,500 to 9,500, 8,000 to 9,500, 8,500 to 9,500, 9,000 to 9,500, 1,500 to 9,000, 2,000 to 9,000, 2,500 to 9,000, 3,000 to 9,000, 3,500 to 9,000, 4,000 to 9,000, 4,500 to 9,000, 5,000 to 9,000, 5,500 to 9,000, 6,000 to 9,000, 6,500 to 9,000, 7,000 to 9,000, 7,500 to 9,000, 8,000 to 9,000, 8,500 to 9,000, 1,500 to 8,500, 2,000 to 8,500, 2,500 to 8,500, 3,000 to 8,500, 3,500 to 8,500, 4,000 to 8,500, 4,500 to 8,500, 5,000 to 8,500, 5,500 to 8,500, 6,000 to 8,500, 6,500 to 8,500, 7,000 to 8,500, 7,500 to 8,500, 8,000 to 8,500, 1,500 to 8,000, 2,000 to 8,000, 2,500 to 8,000, 3,000 to 8,000, 3,500 to 8,000, 4,000 to 8,000, 4,500 to 8,000, 5,000 to 8,000, 5,500 to 8,000, 6,000 to 8,000, 6,500 to 8,000, 7,000 to 8,000, 7,500 to 8,000, 1,500 to 7,500, 2,000 to 7,500, 2,500 to 7,500, 3,000 to 7,500, 3,500 to 7,500, 4,000 to 7,500, 4,500 to 7,500, 5,000 to 7,500, 5,500 to 7,500, 6,000 to 7,500, 6,500 to 7,500, 7,000 to 7,500, 1,500 to 7,000, 2,000 to 7,000, 2,500 to 7,000, 3,000 to 7,000, 3,500 to 7,000, 4,000 to 7,000, 4,500 to 7,000, 5,000 to 7,000, 5,500 to 7,000, 6,000 to 7,000, 6,500 to 7,000, 1,500 to 6,500, 2,000 to 6,500, 2,500 to 6,500, 3,000 to 6,500, 3,500 to 6,500, 4,000 to 6,500, 4,500 to 6,500, 5,000 to 6,500, 5,500 to 6,500, 6,000 to 6,500, 1,500 to 6,000, 2,000 to 6,000, 2,500 to 6,000, 3,000 to 6,000, 3,500 to 6,000, 4,000 to 6,000, 4,500 to 6,000, 5,000 to 6,000, 5,500 to 6,000, 1,500 to 5,500, 2,000 to 5,500, 2,500 to 5,500, 3,000 to 5,500, 3,500 to 5,500, 4,000 to 5,500, 4,500 to 5,500, 5,000 to 5,500, 1,500 to 5,000, 2,000 to 5,000, 2,500 to 5,000, 3,000 to 5,000, 3,500 to 5,000, 4,000 to 5,000, 4,500 to 5,000, 1,500 to 4,500, 2,000 to 4,500, 2,500 to 4,500, 3,000 to 4,500, 3,500 to 4,500, 4,000 to 4,500, 1,500 to 4,000, 2,000 to 4,000, 2,500 to 4,000, 3,000 to 4,000, 3,500 to 4,000, 1,500 to 3,500, 2,000 to 3,500, 2,500 to 3,500, 3,000 to 3,500, 1,500 to 3,000, 2,000 to 3,000, 2,500 to 3,000, 1,500 to 2,500, 2,000 to 2,500, and 1,500 to 2,000 daltons. [0646] In some embodiments the polymer (e.g., PEG) is conjugated to at least one lipid. In some embodiments the lipid conjugated to the polymer comprised of at least one neutral lipid, at least one phospholipid, at least one anionic lipid, at least one cationic lipid, at least one cholesterol, at least one cholesterol derivative, or any combination thereof. [0647] In some embodiments, the lipid conjugated to the polymer may be selected from, but is not limited to, at least one of the cationic, non-cationic, or sterol lipids listed previously. [0648] In some embodiments, the at least one PEG-lipid conjugate may be selected from, but is not limited to at least one of Siglec-1L-PEG-DSPE, R)-2,3-bis(octadecyloxy)propyl-1- (methoxypoly(ethyleneglycol)2000)propylcarbamate, PEG-S-DSG, PEG-S-DMG, PEG-PE, PEG-PAA, PEG-OH DSPE C18, PEG-DSPE, PEG-DSG, PEG-DPG, PEG-DOMG, PEG- DMPE Na, PEG-DMPE, PEG-DMG2000, PEG-DMG C14, PEG-DMG 2000, PEG-DMG, PEG-DMA, PEG-Ceramide C16, PEG-C-DOMG, PEG-c-DMOG, PEG-c-DMA, PEG- cDMA, PEGA, PEG750-C-DMA, PEG400, PEG2k-DMG, PEG2k-C11, PEG2000-PE, PEG2000P, PEG2000-DSPE, PEG2000-DOMG, PEG2000-DMG, PEG2000-C-DMA, PEG2000, PEG200, PEG(2k)-DMG, PEG DSPE C18, PEG DMPE C14, PEG DLPE C12, PEG Click DMG C14, PEG Click C12, PEG Click C10, N(Carbonyl-methoxypolyethylenglycol- 2000)-l,2-distearoyl-sn-glycero3-phosphoethanolamine, Myrj52, mPEG-PLA, MPEG-DSPE, mPEG3000-DMPE, MPEG-2000-DSPE, MPEG2000-DSPE, mPEG2000-DPPE, mPEG2000- DMPE, mPEG2000-DMG, mDPPE-PEG2000, l,2-distearoyl-sn-glycero-3- phosphoethanolamine-PEG2000, HPEG-2K-LIPD, Folate PEG-DSPE, DSPE-PEGMA 500, DSPE-PEGMA, DSPE-PEG6000, DSPE-PEG5000, DSPE-PEG2K-NAG, DSPE-PEG2k, DSPE-PEG2000maleimide, DSPE-PEG2000, DSPE-PEG, DSG-PEGMA, DSG-PEG5000, DPPE-PEG-2K, DPPE-PEG, DPPE-mPEG2000, DPPE-mPEG, DPG-PEGMA, DOPE- PEG2000, DMPE-PEGMA, DMPE-PEG2000, DMPE-Peg, DMPE-mPEG2000, DMG- PEGMA, DMG-PEG2000, DMG-PEG, distearoyl-glycerol-polyethyleneglycol, Cl8PEG750, CI8PEG5000, CI8PEG3000, CI8PEG2000, CI6PEG2000, CI4PEG2000, C18-PEG5000, C18PEG, C16PEG, C16 mPEG (polyethylene glycol) 2000 Ceramide, C14-PEG-DSPE200, C14-PEG2000, C14PEG2000, C14-PEG 2000, C14-PEG, C14PEG, 14:0-PEG2KPE, 1,2- distearoyl-sn-glycero-3-phosphoethanolamine-PEG2000, (R)-2,3-bis(octadecyloxy)propyl-1- (methoxypoly(ethyleneglycol)2000)propylcarbamate, (PEG)-C-DOMG, PEG-C-DMA, and DSPE-PEG-X. [0649] In some embodiments, the LNP comprises a Lipid of the Disclosure, distearoylphosphatidylcholine (DSPC), cholesterol, and 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG2000). [0650] In some embodiments, the LNP comprises a Lipid of the Disclosure, distearoylphosphatidylcholine (DSPC), cholesterol, and 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG2000) at a molar ratio of about 48.5:10:40:1.5, respectively. [0651] In some embodiments, the LNP comprises a Lipid of the Disclosure, distearoylphosphatidylcholine (DSPC), cholesterol, and 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG2000) at a molar ratio of about 48.5:10:39:2.5, respectively. [0652] The amounts and ratios of LNP components may be varied by any amount dependent on the desired form, structure, function, cargo, target, or any combination thereof. The amount of each component may be expressed in various embodiments as percent of the total molar mass of all lipid or lipid conjugated components accounted for by the indicated component (mol%), The amount of each component may be expressed in various embodiments as the relative ratio of each component based on molar mass (Molar Ratio). The amount of each component may be expressed in various embodiments as the weight of each component used to formulate the LNP prior to fabrication (mg or equivalent). The amount of each component may be expressed in various embodiments by any other method known in the art. Any formulation given in one representation of component amounts ("units") is expressly meant to encompass any formulation expressed in different units of component amounts, wherein those representations are effectively equivalent when converted into the same units. In some embodiments, "effectively equivalent" means two or more values within about 10% of one another. [0653] In some embodiments, the LNP comprises at least one cationic lipid in an amount of about 0.1 to 100 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of about 20 to 60 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of about 50 to 85 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of less than about 20 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of more than about 60 mol% or about 85 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of about 95 mol% or less. In some embodiments, the LNP comprises a cationic lipid in an amount of less than or equal to about 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, and 5 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of more than or equal to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, and 95 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount from about 20 to 30 mol%, 20 to 35 mol%, 20 to 40 mol%, 20 to 45 mol%, 20 to 50 mol%, 20 to 55 mol%, 20 to 60 mol%, 20 to 65 mol%, 20 to 70 mol%, 20 to 75 mol%, 20 to 80 mol%, 20 to 85 mol%, 20 to 90 mol%, 25 to 35 mol%, 25 to 40 mol%, 25 to 45 mol%, 25 to 50 mol%, 25 to 55 mol%, 25 to 60 mol%, 25 to 65 mol%, 25 to 70 mol%, 25 to 75 mol%, 25 to 80 mol%, 25 to 85 mol%, 25 to 90 mol%, 30 to 40 mol%, 30 to 45 mol%, 30 to 50 mol%, 30 to 55 mol%, 30 to 60 mol%, 30 to 65 mol%, 30 to 70 mol%, 30 to 75 mol%, 30 to 80 mol%, 30 to 85 mol%, 30 to 90 mol%, 35 to 40 mol%, 35 to 45 mol%, 35 to 50 mol%, 35 to 55 mol%, 35 to 60 mol%, 35 to 65 mol%, 35 to 70 mol%, 35 to 75 mol%, 35 to 80 mol%, 35 to 85 mol%, 35 to 90 mol%, 40 to 45 mol%, 40 to 50 mol%, 40 to 55 mol%, 40 to 60 mol%, 40 to 65 mol%, 40 to 70 mol%, 40 to 75 mol%, 40 to 80 mol%, 40 to 85 mol%, 40 to 90 mol%, 45 to 55 mol%, 45 to 60 mol%, 45 to 65 mol%, 45 to 70 mol%, 45 to 75 mol%, 45 to 80 mol%, 45 to 85 mol%, 45 to 90 mol%, 50 to 60 mol%, 50 to 65 mol%, 50 to 70 mol%, 50 to 75 mol%, 50 to 80 mol%, 50 to 85 mol%, 50 to 90 mol%, 55 to 65 mol%, 55 to 70 mol%, 55 to 75 mol%, 55 to 80 mol%, 55 to 85 mol%, 55 to 90 mol%, 60 to 70 mol%, 60 to 75 mol%, 60 to 80 mol%, 60 to 85 mol%, 60 to 90 mol%, 65 to 75 mol%, 65 to 80 mol%, 65 to 85 mol%, 65 to 90 mol%, 70 to 80 mol%, 70 to 85 mol%, 70 to 90 mol%, 75 to 85 mol%, 75 to 90 mol%, 80 to 90 mol% or 85 to 95 mol%. [0654] In some embodiments, the LNP comprises at least one non-cationic lipid in an amount of about 0.1 to 100 mol%. In some embodiments, the LNP comprises at least one non-one cationic lipid in an amount of about 5 to 35 mol%. In some embodiments, the LNP comprises at least one cationic lipid in an amount of about 5 to 25 mol%. In some embodiments, the LNP comprises at least one non-cationic lipid in an amount of less than about 5 mol%. In some embodiments, the LNP comprises at least one non-cationic lipid in an amount of more than about 25 mol% or about 35 mol%. In some embodiments, the LNP comprises at least one non- cationic lipid in an amount of about 95 mol% or less. In some embodiments, the LNP comprises at least one non-cationic lipid in an amount of less than or equal to about 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, and 5 mol%. In some embodiments, the LNP comprises at least one non-cationic lipid in an amount of more than or equal to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, and 95 mol%. In some embodiments, the LNP comprises at least one non-cationic lipid in an amount from about 5 to 15 mol%, 5 to 25 mol%, 5 to 35 mol%, 5 to 45 mol%, 5 to 55 mol%, 10 to 20 mol%, 10 to 30 mol%, 10 to 40 mol%, 10 to 50 mol%, 15 to 25 mol%, 15 to 35 mol%, 15 to 45 mol%, 20 to 30 mol%, 20 to 40 mol%, 20 to 50 mol%, 25 to 35 mol%, 25 to 45 mol%, 30 to 40 mol%, 30 to 50 mol%, and 35 to 45 mol%. [0655] In some embodiments, the LNP comprises at least one sterol in an amount of about 0.1 to 100 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of about 20 to 45 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of about 25 to 55 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of less than about 20 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of more than about 45 mol% or about 55 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of about 95 mol% or less. In some embodiments, the LNP comprises at least one sterol in an amount of less than or equal to about 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, and 5 mol%. In some embodiments, the LNP comprises at least one sterol in an amount of more than or equal to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, and 95 mol%. In some embodiments, the LNP comprises at least one sterol in an amount from about 10 to 20 mol%, 10 to 30 mol%, 10 to 40 mol%, 10 to 50 mol%, 10 to 60 mol%, 15 to 25 mol%, 15 to 35 mol%, 15 to 45 mol%, 15 to 55 mol%, 15 to 65 mol%, 20 to 30 mol%, 20 to 40 mol%, 20 to 50 mol%, 20 to 60 mol%, 25 to 35 mol%, 25 to 45 mol%, 25 to 55 mol%, 25 to 65 mol%, 30 to 40 mol%, 30 to 50 mol%, 30 to 60 mol%, 35 to 45 mol%, 35 to 55 mol%, 35 to 65 mol%, 40 to 50 mol%, 40 to 60 mol%, 45 to 55 mol%, 45 to 65 mol%, 50 to 60 mol%, and 55 to 65 mol%. [0656] In some embodiments, the LNP comprises at least one particle-activity-modifying- agent in an amount of about 0.1 to 100 mol%. In some embodiments, the LNP comprises at least one particle-activity-modifying-agent in an amount of about 0.5 to 15 mol%. In some embodiments, the LNP comprises at least one particle-activity-modifying-agent in an amount of about 15 to 40 mol%. In some embodiments, the LNP comprises at least one particle- activity-modifying-agent in an amount of less than about 0.1 mol%. In some embodiments, the LNP comprises at least one particle-activity-modifying-agent in an amount of more than about 15 mol% or about 40 mol%. In some embodiments, the LNP comprises at least one particle- activity-modifying-agent in an amount of about 95 mol% or less. In some embodiments, the LNP comprises at least one particle-activity-modifying-agent in an amount of less than or equal to about 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, and 5 mol%. In some embodiments, the LNP comprises at least one particle-activity-modifying-agent in an amount of more than or equal to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, and 95 mol%. In some embodiments, the LNP comprises at least one particle- activity-modifying-agent in an amount from about 0.1 to 1 mol%, 0.1 to 2 mol%, 0.1 to 3 mol%, 0.1 to 4 mol%, 0.1 to 5 mol%, 0.1 to 6 mol%, 0.1 to 7 mol%, 0.1 to 8 mol%, 0.1 to 9 mol%, 0.1 to 10 mol%, 0.1 to 15 mol%, 0.1 to 20 mol%, 0.1 to 25 mol%, 1 to 2 mol%, 1 to 3 mol%, 1 to 4 mol%, 1 to 5 mol%, 1 to 6 mol%, 1 to 7 mol%, 1 to 8 mol%, 1 to 9 mol%, 1 to 10 mol%, 1 to 15 mol%, 1 to 20 mol%, 1 to 25 mol%, 2 to 3 mol%, 2 to 4 mol%, 2 to 5 mol%, 2 to 6 mol%, 2 to 7 mol%, 2 to 8 mol%, 2 to 9 mol%, 2 to 10 mol%, 2 to 15 mol%, 2 to 25 mol%, 3 to 4 mol%, 3 to 5 mol%, 3 to 6 mol%, 3 to 7 mol%, 3 to 8 mol%, 3 to 9 mol%, 3 to 10 mol%, 3 to 15 mol%, 3 to 20 mol%, 3 to 25 mol%, 4 to 5 mol%, 4 to 6 mol%, 4 to 7 mol%, 4 to 8 mol%, 4 to 9 mol%, 4 to 10 mol%, 4 to 15 mol%, 4 to 20 mol%, 4 to 25 mol%, 5 to 10 mol%, 5 to 15 mol%, 5 to 20 mol%, 5 to 25 mol%, 10 to 15 mol%, 10 to 20 mol%, 10 to 25 mol%, 15 to 20 mol%, 15 to 25 mol%, and 20 to 25 mol%. [0657] In some embodiments, the LNP is comprised of about 30-60 mol% of at least one cationic lipid, about 0-30 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 18.5-48.5 mol% of at least one sterol (e.g., cholesterol), and about 0-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0658] In some embodiments, the LNP is comprised of about 35-55 mol% of at least one cationic lipid, about 5-25 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 30-40 mol% of at least one sterol (e.g., cholesterol), and about 0-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0659] In some embodiments, the LNP is comprised of about 35-45 mol% of at least one cationic lipid, about 25-35 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 20-30 mol% of at least one sterol (e.g., cholesterol), and about 0-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0660] In some embodiments, the LNP is comprised of about 45-65 mol% of at least one cationic lipid, about 5-10 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 25-40 mol% of at least one sterol (e.g., cholesterol), and about 0.5-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0661] In some embodiments, the LNP is comprised of about 40-60 mol% of at least one cationic lipid, about 5-15 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 35-45 mol% of at least one sterol (e.g., cholesterol), and about 0.5-3 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0662] In some embodiments, the LNP is comprised of about 30-60 mol% of at least one cationic lipid, about 0-30 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 15-50 mol% of at least one sterol (e.g., cholesterol), and about 0.01-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0663] In some embodiments, the LNP is comprised of about 10-75 mol% of at least one cationic lipid, about 0.5-50 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 5-60 mol% of at least one sterol (e.g., cholesterol), and about 0.1-20 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0664] In some embodiments, the LNP is comprised of about 50-65 mol% of at least one cationic lipid, about 3-15 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 30-40 mol% of at least one sterol (e.g., cholesterol), and about 0.5-2 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0665] In some embodiments, the LNP is comprised of about 50-85 mol% of at least one cationic lipid, about 3-15 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 30-40 mol% of at least one sterol (e.g., cholesterol), and about 0.5-2 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0666] In some embodiments, the LNP is comprised of about 25-75 mol% of at least one cationic lipid, about 0.1-15 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 5-50 mol% of at least one sterol (e.g., cholesterol), and about 0.5-20 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0667] In some embodiments, the LNP is comprised of about 50-65 mol% of at least one cationic lipid, about 5-10 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 25-35 mol% of at least one sterol (e.g., cholesterol), and about 5-10 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0668] In some embodiments, the LNP is comprised of about 20-60 mol% of at least one cationic lipid, about 5-25 mol% of at least one non-cationic lipid (e.g., a phospholipid), about 25-55 mol% of at least one sterol (e.g., cholesterol), and about 0.5-15 mol% of at least one particle-activity-modifying-agent (e.g., a PEGylated lipid). [0669] In some embodiments, the LNPs can be characterized by their shape. In some embodiments, the LNPs are essentially spherical. In some embodiments, the LNPs are essentially rod-shaped (i.e., cylindrical). In some embodiments, the LNPs are essentially disk shaped. [0670] In some embodiments, the LNPs can be characterized by their size. In some embodiments, the size of an LNP can be defined as the diameter of its largest circular cross section, referred to herein simply as its diameter. In some embodiments the LNPs may have a diameter between 30 nm to about 150 nm. In some embodiments, the LNP may have diameters ranging between about 40 to 150 nm 50 to 150 nm, 60 to 150 nm, about 70 to 150 nm, or 80 to 150 nm, 90 to 150 nm, 100 to nm, 110 to 150 nm, 120 to 150 nm, 130 to 150 nm, 140 to 150 nm, 30 to 30 to 140 mol%, 40 to 140 mol%, 50 to 140 mol%, 60 to 140 mol%, 70 to 140 mol%, 80 to 140 mol%, 90 to 140 mol%, 100 to 140 mol%, 110 to 140 mol%, 120 to 140 mol%, 130 to 140 mol%, 140 to 140 mol%, 30 to 140 mol%, 40 to 130 mol%, 50 to 130 mol%, 60 to 130 mol%, 70 to 130 mol%, 80 to 130 mol%, 90 to 130 mol%, 100 to 130 mol%, 110 to 130 mol%, 120 to 130 mol%, 30 to 120 mol%, 40 to 120 mol%, 50 to 120 mol%, 60 to 120 mol%, 70 to 120 mol%, 80 to 120 mol%, 90 to 120 mol%, 100 to 120 mol%, 110 to 120 mol%, 30 to 110 mol%, 40 to 110 mol%, 50 to 110 mol%, 60 to 110 mol%, 70 to 110 mol%, 80 to 110 mol%, 90 to 110 mol%, 100 to 110 mol%, 30 to 100 mol%, 40 to 100 mol%, 50 to 100 mol%, 60 to 100 mol%, 70 to 100 mol%, 80 to 100 mol%, 90 to 100 mol%, 30 to 90 mol%, 40 to 90 mol%, 50 to 90 mol%, 60 to 90 mol%, 70 to 90 mol%, 80 to 90 mol%, 30 to 80 mol%, 40 to 80 mol%, 50 to 80 mol%, 60 to 80 mol%, 70 to 80 mol%, 30 to 70 mol%, 40 to 70 mol%, 50 to 70 mol%, 60 to 70 mol%, 30 to 60 mol%, 40 to 60 mol%, 50 to 60 mol%, 30 to 50 mol%, 40 to 50 mol%, and 30 to 40 mol%. [0671] In some embodiments, a population of LNPs, such as those resulting from the same formulation, may be characterized by measuring the uniformity of size, shape, or mass of the particles in the population. Uniformity may be expressed in some embodiments as the polydispersity index (PI) of the population. In some embodiments uniformity may be expressed in some embodiments as the disparity (Đ) of the population. The terms "polydispersity index" and "disparity" are understood herein to be equivalent and may be used interchangeably. In some embodiments, a population of LNPs resulting from a given formulation will have a PI of between about 0.1 and 1. In some embodiments, a population of LNPs resulting from a giving formulation will have a PI of less than about 1, less than about 0.5, less than about 0.4, less than about 0.3, less than about 0.2, less than about 0.1. In some embodiments, a population of LNPs resulting from a given formulation will have a PI of between about 0.1 to 1, 0.1 to 0.8, 0.1 to 0.6, 0.1 to 0.4, 0.1 to 0.2, 0.2 to 1, 0.2 to 0.8, 0.2 to 0.6, 0.2 to 0.4, 0.4 to 1, 0.4 to 0.8, 0.4 to 0.6, 0.6 to 1, 0.6 to 0.8, and 0.8 to 1. [0672] In some embodiments, the LNP may fully or partially encapsulate a cargo, such as the originator constructs and benchmark constructs of the present disclosure. In some embodiments, essentially 0% of the cargo present in the final formulation is exposed to the environment outside of the LNP (i.e., the cargo is fully encapsulated. In some embodiments, the cargo is associated with the LNP but is at least partially exposed to the environment outside of the LNP. In some embodiments, the LNP may be characterized by the % of the cargo not exposed to the environment outside of the LNP, e.g., the encapsulation efficiency. For the sake of clarity, an encapsulation efficiency of about 100% refers to an LNP formulation where essentially all the cargo is fully encapsulated by the LNP, while an encapsulation rate of about 0% refers to an LNP where essential none of the cargo is encapsulated in the LNP, such as with an LNP where the cargo is bound to the external surface of the LNP. On some embodiments, an LNP may have an encapsulation efficiency of less than about 100%, less than about 95%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15% less than about 10%, or less than 5%. In some embodiments, an LNP may have an encapsulation efficiency of between about 90 to 100%, 80 to 100%, 70 to 100%, 60 to 100%, 50 to 100%, 40 to 100%, 30 to 100%, 20 to 100%, 10 to 100%, 80 to 90%, 70 to 90%, 60 to 90%, 50 to 90%, 40 to 90%, 30 to 90%, 20 to 90%, 10 to 90%, 70 to 80%, 60 to 80%, 50 to 80%, 40 to 80%, 30 to 80%, 20 to 80%, 10 to 80%, 60 to 70%, 50 to 70%, 40 to 70%, 30 to 70%, 20 to 70%, 10 to 70%, 40 to 50%, 30 to 50%, 20 to 50%, 10 to 50%, 30 to 40%, 20 to 40%, 10 to 40%, 20 to 30%, 10 to 30%, and 10 to 20%. [0673] In some embodiments, a LNP may include at least one identifier moiety as shown in FIG. 5. Non-limiting examples of an identifier moiety include glycans, antibodies, peptides, small molecules, and any combination thereof. In some embodiments, the at least one targeting agent may be incorporated into the lipid membrane of the lipid-based nanoparticle. In some embodiments, the at least one targeting agent may be presented on the external surface of the nanoparticle. In some embodiments, the at least one targeting agent may be conjugated to a lipid-component of the nanoparticle. In some embodiments, the at least one targeting agent may be conjugated to a polymer component of the nanoparticle. In some embodiments, the at least one targeting agent may be anchored to the nanoparticle via hydrophobic ad hydrophilic interactions among the at least one targeting agent, the nanoparticle membrane, and the aqueous environments inside or outside the nanoparticle. In some embodiments, the at least one targeting agent is conjugated to a peptide/protein component of the nanoparticle membrane. In some embodiments, the at least one targeting agent is conjugated to a suitable linker moiety which is conjugated to a component of the nanoparticle membrane. In some embodiments, any combination of forces and bonds can result in the targeting agent being associated with the nanoparticle. [0674] The LNPs described herein may be formed using techniques known in the art. As a non- limiting example, an organic solution containing the lipids is mixed together with an acidic aqueous solution containing the originator construct or benchmark construct in a microfluidic channel resulting in the formation of targeting system (delivery vehicle and the benchmark construct). [0675] In some embodiments, each LNP formulation includes a benchmark construct having a uniquely identifiable nucleotide identifier sequence (e.g., barcode). The unique identifier sequence provides the ability to identify the specific LNP which produces the desired result. The LNP formulation may also differ in the LNP-forming composition used to generate the LNP. For example, the LNP-forming compositions can be varied in the molar amount and/or structure of the ionizable lipid, the molar amount and/or structure of the helper lipid, the molar amount/or structure of PEG, and/or the molar amount of cholesterol. Additionally, or alternatively, the LNP formulation may comprise benchmark constructs which differ in the coding sequence for the biologically active molecule. Additionally, or alternatively, the LNP formulation may comprise benchmark constructs which differ in the modifications made to the nucleic acid sequence. [0676] In some embodiments, the lipid compositions described according to the respective molar ratios of the component lipids in the formulation. As a non-limiting example, the mol-% of the ionizable lipid may be from about 10 mol-% to about 80 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 20 mol-% to about 70 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 30 mol-% to about 60 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 35 mol-% to about 55 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 40 mol-% to about 50 mol-%. As a non-limiting example, the ionizable lipid mol-% of the transfer vehicle batch will be ±30%, ±25%, ±20%, ±15%, ±10%, ±5%, or ±2.5% of the target mol-%. In some embodiments, transfer vehicle variability between lots will be less than 15%, less than 10% or less than 5%. [0677] In some embodiments, the mol-% of the helper lipid may be from about 1 mol-% to about 50 mol-%. In some embodiments, the mol-% of the helper lipid may be from about 2 mol-% to about 45 mol-%. In some embodiments, the mol-% of the helper lipid may be from about 3 mol-% to about 40 mol-%. In some embodiments, the mol-% of the helper lipid may be from about 4 mol-% to about 35 mol-%. In some embodiments, the mol-% of the helper lipid may be from about 5 mol-% to about 30 mol-%. In some embodiments, the mol-% of the helper lipid may be from about 10 mol-% to about 20 mol-%. In some embodiments, the helper lipid mol-% of the transfer vehicle batch will be ±30%, ±25%, ±20%, ±15%, ±10%, ±5%, or ±2.5% of the target mol-%. [0678] In some embodiments, the mol-% of the structural lipid may be from about 10 mol-% to about 80 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 20 mol-% to about 70 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 30 mol-% to about 60 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 35 mol-% to about 55 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 40 mol-% to about 50 mol-%. In some embodiments, the structural lipid mol-% of the transfer vehicle batch will be ±30%, ±25%, ±20%, ±15%, ±10%, ±5%, or ±2.5% of the target mol-%. [0679] In some embodiments, the mol-% of the PEG modified lipid may be from about 0.1 mol-% to about 10 mol-%. In some embodiments, the mol-% of the PEG modified lipid may be from about 0.2 mol-% to about 5 mol-%. In some embodiments, the mol-% of the PEG modified lipid may be from about 0.5 mol-% to about 3 mol-%. In some embodiments, the mol-% of the PEG modified lipid may be from about 1 mol-% to about 2 mol-%. In some embodiments, the mol-% of the PEG modified lipid may be about 1.5 mol-%. In some embodiments, the PEG modified lipid mol-% of the transfer vehicle batch will be ±30%, ±25%, ±20%, ±15%, ±10%, ±5%, or ±2.5% of the target mol-%. [0680] In some embodiments, the delivery vehicle may be any of the lipid nanoparticles described in WO2021113777, the contents of which are herein incorporated by reference in their entirety. [0681] In some embodiments, the delivery vehicle is a lipid nanoparticle which comprises any of the ionizable lipids (e.g., amine lipids), PEG lipids, non-cationic (helper) lipids, or structural lipids in WO2021113777, the contents of which are herein incorporated by reference in their entirety. [0682] In some embodiments, a lipid nanoparticle formulation may be prepared by the methods described in International Publication Nos. WO2011127255 or W02008103276, the contents of each of which is herein incorporated by reference in their entirety. In some embodiments, lipid nanoparticle formulations may be as described in International Publication No. W02019131770, the contents of which is herein incorporated by reference in its entirety. [0683] In some embodiments, a lipid nanoparticle formulation may be prepared by the methods described in International Publication No. WO2020237227, the contents of each of which is herein incorporated by reference in their entirety. In some embodiments, lipid nanoparticle formulations may be as described in International Publication No. WO2020237227, the contents of which is herein incorporated by reference in its entirety. Non-Lipid Nanoparticle [0684] In some embodiments, the nanoparticle is a non-lipid-based nanoparticle. Non-lipid- based nanoparticles include, but are not limited to, silicon-based nanoparticles (i.e., porous silicon nanoparticles), gold nanoparticles, polypeptide-based nanoparticles, nucleotide-based nanoparticles, and carbon-based nanoparticle. Exosomes [0685] In some embodiments, the delivery vehicle comprises at least one exosome. As used herein, "exosomes" refer to small membrane bound vesicles with an endocytic origin. Without wishing to be bound by theory, exosomes are generally released into an extracellular environment from host/progenitor cells post fusion of multivesicular bodies the cellular plasma membrane. As such, exosomes will tend to include components of the progenitor membrane in addition to designed components and cargos. Exosome membranes are generally lamellar, composed of a bilayer of lipids, with an aqueous inter-nanoparticle space. [0686] In some embodiments, an exosome may include at least one identifier moiety as shown in FIG.5. Non-limiting examples of an identifier moiety include glycans, antibodies, peptides, small molecules, and any combination thereof. Liposomes [0687] In some embodiments, the delivery vehicles comprise of at least one liposome. As used herein, "liposomes" are small vesicles comprised of at least one lipid bilayer membrane surrounding an aqueous inner-nanoparticle space that is generally not derived from a progenitor/host cell. Liposomes can be (large) multilamellar vesicle (MLV), potentially hundreds of nanometers in diameter comprising a series of concentric bilayers separated by narrow aqueous spaces, small unicellular vesicle (SUV), potentially smaller than 50 nm in diameter, and a large unilamellar vesicle (LUV), potentially between 50 and 500 nm in diameter. In some embodiments, liposomes may be comprised of any or all the same components and same component amounts as a lipid nanoparticle, differing principally in their method of manufacture. Micelles [0688] In some embodiments, the delivery vehicles comprise of at least one micelle. As used herein, "micelles" refer to small particles which do not have an aqueous intra-particle space. Without wishing to be bound by theory, the intra-particle space of micelles is occupied by the hydrophobic tails of the lipids comprising the micelle membrane and possible associated cargo, rather than any additional lipid-head groups. In some embodiments, micelles may be comprised of any or all the same components as a lipid-nanoparticle, differing principally in their method of manufacture. [0689] In some embodiments, a micelle may include at least one identifier moiety as shown in FIG. 5. Non-limiting examples of an identifier moiety include glycans, antibodies, peptides, small molecules, and any combination thereof. Viral particle [0690] In some embodiments, the delivery vehicle comprises at least one virus like particle. As used herein, "virus like particles" refer to a vesicle predominantly of a protein capsid, sheath, shell, or coat (all used interchangeably herein) derived from a virus which can be loaded with a cargo moiety. In general, virus like particle are non-infection and may be synthesized using cellular machinery to express viral capsid protein sequences, which then self-assemble and incorporate the associated cargo moiety, though it is possible to form virus like particles by providing the capsid and cargo components without expression related cellular machinery and allowing them to self-assemble. [0691] In some embodiments, the virus like particle may be derived from at least one of species of virus such as, but not limited to, Parvoviridae, Retroviridae, Flaviviridae, Paramyxoviridae, and bacteriophages. In some embodiments, the virus like particle may be derived from an adeno-associated virus, HIV, Hepatitis C virus, HPV, or any combination thereof. [0692] In some embodiments, the virus like particle is an AAV particle and the AAV particle may include at least one identifier moiety as shown in FIG. 5. Non-limiting examples of an identifier moiety include glycans, antibodies, peptides, small molecules, and any combination thereof. Polymeric delivery [0693] In some embodiments, the delivery vehicle may comprise at least one polymeric delivery agent. As used herein, "polymeric delivery agents" refer to non-aggregating delivery agents comprised of soluble polymers conjugated to cargo moieties via various linkage groups. In some embodiments, polymeric delivery agents may comprise any of the polymers described herein. Tracking Systems [0694] The tropism discovery platform disclosed herein may utilize a variety of tracking systems which include identifier sequences and moieties (also referred to as a "barcode") in order to allow qualification of the delivery vehicles and/or the benchmark constructs, cargo and payloads post-administration. [0695] In some embodiments, the tracking system is a single identifier sequence or moiety. The identifier sequence or moiety may be located in the delivery vehicle, benchmark construct, cargo or payload region, 5' UTR, 3'UTR, promoter region or tailing region. As a non-limiting example, the identifier sequence or moiety is located in or on the delivery vehicle. As a non- limiting example, the identifier sequence or moiety is located in or on the benchmark construct. As a non-limiting example, the identifier sequence or moiety is located in or on the 5' UTR. As a non-limiting example, the identifier sequence or moiety is located in or on the 3' UTR. As a non-limiting example, the identifier sequence or moiety is located in or on the promoter region. As a non-limiting example, the identifier sequence or moiety is located in or on the payload region. As a non-limiting example, the identifier sequence or moiety is located in or on the tailing region. [0696] In some embodiments, the tracking system is a set of identifier sequences or moieties with a first identifier sequence or moiety for the delivery vehicle and a second identifier sequence or moiety for the benchmark construct, cargo and payload. The first and second identifier sequence or moiety may be the same or different. If there are additional benchmark constructs, cargos and payloads in the delivery vehicle then each benchmark constructs, cargo and payloads may have its own identifier sequence or moiety or it may be the same at the second identifier sequence or moiety. [0697] In some embodiments, the tropism discovery platform is comprised of multiple tracking systems, wherein each tracking system allows for detecting the delivery vehicle and/or benchmark constructs, cargo and payloads at different levels of resolution. [0698] In some embodiments, the tracking systems comprises at least one barcode sequence. As used herein, a "barcode" or "barcode sequence" is any sequence which can be detected using methods known in the art and is distinct from the sequences in the cell, tissue, organ and/or organism or any sequences being administered. The barcode sequence may be included in or attached to the delivery vehicle and/or in the benchmark construct, cargo and payload. As a non-limiting example, the delivery vehicle comprises the barcode sequence. As a non-limiting example, the cargo or payload comprises the barcode sequence. As a non-limiting example, the benchmark construct comprises the barcode sequence. [0699] In some embodiments, the location of the identifier sequence or moiety in the targeting system is random. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct and the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle, benchmark construct, and the cargo or payload. [0700] In some embodiments, the location of the identifier sequence or moiety in the targeting system is pre-determined. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct and the cargo or payload. As a non- limiting example, the identifier sequence or moiety is in the delivery vehicle, benchmark construct, and the cargo or payload. [0701] In some embodiments, the location of the identifier sequence or moiety in the targeting system is inverted. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the benchmark construct. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle and the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the benchmark construct and the cargo or payload. As a non-limiting example, the identifier sequence or moiety is in the delivery vehicle, benchmark construct, and the cargo or payload. [0702] In some embodiments, the identifier sequence is a randomly generated sequences which serve to avoid duplication during deep sequencing. In some embodiments, the identifier sequence is a repeating sequence of nucleotides or amino acids. In some embodiments, the identifier sequence is a fragment of a larger sequence such as, but not limited to, a cargo or payload. The identifier sequence may be designed to any length available using synthesis technology (See Clement et al., AmpUMI: design and analysis of unique molecular identifiers for deep amplicon sequencing, Bioinformatics, Volume 34, Issue 13, 01 July 2018, Pages i202- i210; the contents of which is herein incorporated herein by reference in its entirety). [0703] In some embodiments, the identifier sequence has a length between 2 and 1000 nucleotides. For example, the identifier sequence may have a length of 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000 or more than 1000 nucleotides. The identifier sequence may have a length between 2-5, 2-10, 2-15, 2-20, 2-30, 2-50, 2-70, 2-90, 2-100, 2-250, 2-300, 2-350, 2-400, 2-450, 2-500, 2-550, 2-600, 2-650, 2-700, 2-750, 2-800, 2-850, 2-900, 2-950, 2-1000, 5-10, 5-15, 5-20, 5-30, 5-50, 5-70, 5-90, 5-100, 5-250, 5-300, 5-350, 5-400, 5-450, 5-500, 5-550, 5-600, 5-650, 5-700, 5-750, 5-800, 5-850, 5-900, 5-950, 5-1000, 10-30, 10-50, 10-70, 10-90, 10-100, 10-250, 10- 300, 10-350, 10-400, 10-450, 10-500, 10-550, 10-600, 10-650, 10-700, 10-750, 10-800, 10- 850, 10-900, 10-950, 10-1000, 20-30, 20-50, 20-70, 20-90, 20-100, 20-250, 20-300, 20-350, 20-400, 20-450, 20-500, 20-550, 20-600, 20-650, 20-700, 20-750, 20-800, 20-850, 20-900, 20- 950, 20-1000, 30-50, 30-70, 30-90, 30-100, 30-250, 30-300, 30-350, 30-400, 30-450, 30-500, 30-550, 30-600, 30-650, 30-700, 30-750, 30-800, 30-850, 30-900, 30-950, 30-1000, 40-50, 40- 70, 40-90, 40-100, 40-250, 40-300, 40-350, 40-400, 40-450, 40-500, 40-550, 40-600, 40-650, 40-700, 40-750, 40-800, 40-850, 40-900, 40-950, 40-1000, 50-70, 50-90, 50-100, 50-250, 50- 300, 50-350, 50-400, 50-450, 50-500, 50-550, 50-600, 50-650, 50-700, 50-750, 50-800, 50- 850, 50-900, 50-950, 50-1000, 60-70, 60-90, 60-100, 60-250, 60-300, 60-350, 60-400, 60-450, 60-500, 60-550, 60-600, 60-650, 60-700, 60-750, 60-800, 60-850, 60-900, 60-950, 60-1000, 70-90, 70-100, 70-250, 70-300, 70-350, 70-400, 70-450, 70-500, 70-550, 70-600, 70-650, 70- 700, 70-750, 70-800, 70-850, 70-900, 70-950, 70-1000, 80-90, 80-100, 80-250, 80-300, 80- 350, 80-400, 80-450, 80-500, 80-550, 80-600, 80-650, 80-700, 80-750, 80-800, 80-850, 80- 900, 80-950, 80-1000, 90-100, 90-250, 90-300, 90-350, 90-400, 90-450, 90-500, 90-550, 90- 600, 90-650, 90-700, 90-750, 90-800, 90-850, 90-900, 90-950, 90-1000, 100-250, 100-300, 100-350, 100-400, 100-450, 100-500, 100-550, 100-600, 100-650, 100-700, 100-750, 100-800, 100-850, 100-900, 100-950, 100-1000, 150-250, 150-300, 150-350, 150-400, 150-450, 150- 500, 150-550, 150-600, 150-650, 150-700, 150-750, 150-800, 150-850, 150-900, 150-950, 150-1000, 200-250, 200-300, 200-350, 200-400, 200-450, 200-500, 200-550, 200-600, 200- 650, 200-700, 200-750, 200-800, 200-850, 200-900, 200-950, 200-1000, 250-300, 250-350, 250-400, 250-450, 250-500, 250-550, 250-600, 250-650, 250-700, 250-750, 250-800, 250-850, 250-900, 250-950, 250-1000, 300-350, 300-400, 300-450, 300-500, 300-550, 300-600, 300- 650, 300-700, 300-750, 300-800, 300-850, 300-900, 300-950, 300-1000, 350-400, 350-450, 350-500, 350-550, 350-600, 350-650, 350-700, 350-750, 350-800, 350-850, 350-900, 350-950, 350-1000, 400-450, 400-500, 400-550, 400-600, 400-650, 400-700, 400-750, 400-800, 400- 850, 400-900, 400-950, 400-1000, 450-500, 450-550, 450-600, 450-650, 450-700, 450-750, 450-800, 450-850, 450-900, 450-950, 450-1000, 500-550, 500-600, 500-650, 500-700, 500- 750, 500-800, 500-850, 500-900, 500-950, 500-1000, 550-600, 550-650, 550-700, 550-750, 550-800, 550-850, 550-900, 550-950, 550-1000, 600-650, 600-700, 600-750, 600-800, 600- 850, 600-900, 600-950, 600-1000, 650-700, 650-750, 650-800, 650-850, 650-900, 650-950, 650-1000, 700-750, 700-800, 700-850, 700-900, 700-950, 700-1000, 750-800, 750-850, 750- 900, 750-950, 750-1000, 800-850, 800-900, 800-950, 800-1000, 850-900, 850-950, 850-1000, 900-950, 900-1000, 950-1000 or over 1000 nucleotides. [0704] In some embodiments, the identifier sequence or moiety may produce a signal that is detectable immediately after administration. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable for an indefinite amount of time after administration. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable for more than 1 day, 2 days, 3 days, 4 days, 5 days, 6 days or 7 days post administration. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable for about 1 to 24 hours. As a non-limiting example, the signal may be detectable for about 1 to 6, 1 to 12, 1 to 18, 6 to 12, 6 to 18, 6 to 24, or 18 to 24 hours, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable for about 1-60 minutes such as, but not limited to, 1-5, 1-10, 1-20, 1-30, 1-40, 1-50, 10-20, 10-30, 10-40, 10-50, 10-60, 20-30, 20-40, 20-50, 20-60, 30-40, 30-50, 30-60, 40-50, 40-60, or 50-60 minutes, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 minutes. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable for less than 1 minute post administration. [0705] In some embodiments, the identifier sequence or moiety may produce a signal that is detectable from outside the body of a subject. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable from via non-invasive imagery techniques, for example from outside a subject's organs or tissues but within the subject's body. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable on a macroscopic level. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable on the microscopic level. In some embodiments, the identifier sequence or moiety may produce a signal that is detectable on the nanoscopic level. In some embodiments, the identifier sequence or moiety may produce a signal that is only detectable after target cells are harvested and assayed, for non-limiting example via mass spectrometer, electrophoresis, flow cytometry, or deep sequencing. [0706] In some embodiments, the delivery vehicle comprises or is operably linked to an identifier moiety. [0707] In some embodiments, the delivery vehicle comprises or is operably linked to an identifier moiety that binds to an immune cell antigen. As a non-limiting example, the immune cell antigen may be a T cell antigen such as CD2, CD3, CD5, CD7, CD8, CD4, beta 7 integrin, beta 2 integrin, and C1q. As a non-limiting example, the immune cell antigen may be a NK cell, an NKT cell, a macrophage or a neutrophil. As a non-limiting example, the immune cell antigen may be a macrophage antigen such as mannose receptor, CD206 and C1q. [0708] In some embodiments, the delivery vehicle comprises or is operably linked to an identifier moiety which is a small molecule that binds to an ectoenzyme on an immune cell. The ectoenzyme may be, but is not limited to, CD38, CD73, adenosine 2a receptor and adenosine 2b receptor. [0709] In some embodiments, the delivery vehicle comprises or is operably linked to an identifier moiety which is a small molecule such as, but not limited to, mannose, lectin, acivicin, biotin, or digoxigenin. [0710] In some embodiments, the delivery vehicle comprises or is operably linked to an identifier moiety which is a single chain Fv (scFv) fragment, nanobody, peptide, peptide-based macrocycle, minibody, small molecule ligand (e.g., folate, arginylglycylaspartic acid (RGD), or phenol-soluble modulin alpha 1 peptide (PSMA1)), heavy chain variable region, light chain variable region or fragment thereof. Tracking System: Fluorescence [0711] In some embodiments, the at least one tracking system comprises an identifier sequence or moieties that is detectable by florescence. [0712] In some embodiments, florescence is achieved via the inclusion of at least one fluorescent dye in the delivery vehicle. In some embodiments, the at least one fluorescent dye may be selected from, but is not limited to, fluorescein, TAMRA (carboxytetramethylrhodamine), Cy dyes, Texas red, HEX, JOE, Oregon green, rhodamine 6 G, coumarin, pyrene, and DiOC6 (3,3′-dihexyloxacarbocyanine iodide). [0713] In some embodiments, florescence is achieved via the inclusion of at least one fluorescent protein in the, or associated with, the delivery vehicle. In some embodiments, at least one fluorescent protein is encoded in the benchmark construct or the benchmark construct comprises the fluorescent protein. Non-limiting examples of fluorescent protein include Green Fluorescent Protein (GFP), Yellow Fluorescent Protein (YFP), Red Fluorescent Protein (RFP), Sirius, excitable blue fluorescent protein (EBFP2), cyan fluorescent protein (CFP), Cerulean, excitable green fluorescent protein (EGFP), excitable yellow fluorescent protein (EYFP), mOrange, mCherry, mPlum, NIR, iRFP, EosFP, PamCherry, Dronpa, Dreiklang, asFP595, mMaple, mGeo, mEos2, Dendra2, psCFP2, and 2,3,5,6-tetracarbazole-4-cyano-pyridine (CPy). [0714] In some embodiments, florescence is achieved via the inclusion of at least one fluorescent nanoparticle associated with the delivery vehicle or the benchmark construct. In some embodiments, the fluorescent nanoparticle may be, but is not limited to, carbon dots, graphene quantum dots, gold nanorods, polymer-based nanoparticles, aggregation-induced emission dots, Conjugated Polymer nanoparticles (CP-dots), Gold nanospheres, Gold nano shells, Gold nanocages, and AIE pheromone. [0715] In some embodiments, florescence is achieved via inclusion of at least one fluorescent lipid associated with or included in the delivery vehicle. In some embodiments, the fluorescent lipid may be, but is not limited to, DiR, DiD, DiO, and DiI, other members of the Di series of phospholipids, Bodipy, and FL-Sphingomyelin. [0716] In some embodiments, florescence is achieved via the inclusion of at least one luciferase in or associated with the delivery vehicle. In some embodiments, at least one luciferase protein is encoded in the benchmark construct or the benchmark construct comprises the luciferase. Non-limiting examples of the types of luciferase which may be used include Renilla luciferase, Gaussia luciferase, Nanoluc luciferase, Firefly luciferase, and Click Beetle luciferases. [0717] In some embodiments, florescence is achieved via inclusion of β-galactosidase (β-gal) associated with or included in the delivery vehicle. In some embodiments, at least one β- galactosidase (β-gal) protein is encoded in the benchmark construct or the benchmark construct comprises β-galactosidase (β-gal). [0718] In some embodiments, florescence is achieved via inclusion of at least one quencher molecule associated with or included in the delivery vehicle. In some embodiments, florescence is achieved via inclusion of at least one quencher molecule associated with or encoded by the benchmark construct. Non-limiting examples of quencher molecules include dimethylaminophenylazobenzoic acid (DABCYL), QSY 7, Cu(II) ion, Dabcyl, QSY 35, BHQ- 0, Eclipse, BHQ-1, QSY 9, BHQ-2, ElleQuencher, Iowa Black, QSY 21, and BHQ-3. Tracking System: Fluorophores and Radioactive Phosphates [0719] In some embodiments, the at least one tracking system comprises an identifier sequence or moieties that is a fluorophore or radioactive phosphate. [0720] In some embodiments, the at least one tracking system comprises the inclusion of at least one fluorophore associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one fluorophore associated with, encoded in or included in the benchmark construct. Non-limiting examples of fluorophores includes quantum dot and organic small molecule. [0721] In some embodiments, the at least one tracking system comprises the inclusion of at least one quantum dot associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one quantum dot associated with, encoded in or included in the benchmark construct. Non-limiting examples of quantum dots include CdSe/ZnS, CdTe/ZnS, CdTe/CdSe, CdSe/ZnTe, CdSe/CdTe/ZnSe, nAs/ZnSe, InAs/CdSe, InAs/InP, Cu:InP/ZnSe, InAsxP1–x/InP/ZnSe, CdS/CdSe, ZnSe/CdSe, ZnSe/InP/ZnS, ZnSe/InP/ZnS, CdTe/ZnSe, QD585, and QD655. [0722] In some embodiments, the at least one tracking system comprises the inclusion of at least one organic small molecule associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one organic small molecule associated with, encoded in or included in the benchmark construct. Non- limiting examples of organic small molecules include classes of Coumarins, Naphthalimides, Fluoresceins and rhodamines derivatives, BODIPY, Cyanines, xanthenes, oxazines, Oligothiophenes, and Phthalocyanine derivatives (PcDer). In some embodiments, the at least one organic small molecule may be selected from, but is not limited to, 7-dialkyl-amino-4- trifluoromethyl coumarin, rhodamine B, Coumarin 314, Lucifer Yellow CH, florescein, rhodamine 123, BODIPY FL NHS ester, Cy5, Rhodamine 6G, Silicon-rhodamine (SiR), Cy3, Cy5.5, Cy7, Cy2, ATTO655, ATTO680, ATTO700, Nitrobenzoxadiazole (NBD), 1,6- diphenyl-1,3,5-hexatriene (DPH), ABBERIOR™, ALEXA FLUOR™, ATTO™, DYLIGHT FLUOR™, ALEXA FLUOR 647™, and TOPFLUOR™. [0723] In some embodiments, the at least one tracking system comprises the inclusion of at least one imaging contrast agent associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one imaging contrast agent associated with, encoded in or included in the benchmark construct. Non- limiting examples of imaging contrast agents include gadolinium-based small molecules, gadolinium-encapsulated liposomes, manganese-based small molecules, and iron oxide nanoparticles. [0724] In some embodiments, the at least one tracking system comprises the inclusion of at least one radiolabel associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one radiolabel associated with, encoded in or included in the benchmark construct. Non-limiting examples of radiolabels include 111 In, 99m Tc, 13 N, 68 Ga, 18 F, 64 Cu, 86 Y, 76 Br, 89 Zr, 72 As, 124 I, 74 As, fluorine-18, gallium- 68, nitrogen-13, copper-64, bromine-76, iodine-125, arsenic-74, carbon-11, iodine-131, 153 Sm, 177 Lu, 186 Re, 188 Re, 198 Au, and 225 Ac. [0725] In some embodiments, the at least one tracking system comprises the inclusion of at least one biotin associated with or included in the delivery vehicle. [0726] In some embodiments, the at least one tracking system comprises the inclusion of at least one digoxygenin associated with or included in the delivery vehicle. [0727] In some embodiments, the at least one tracking system comprises the inclusion of at least one dinitrophenyl (DNP) associated with or included in the delivery vehicle. [0728] In some embodiments, the at least one tracking system comprises the inclusion of at least one Fluorescein associated with or included in the delivery vehicle. [0729] In some embodiments, the at least one tracking system comprises the inclusion of at least one fucose associated with or included in the delivery vehicle. [0730] In some embodiments, the at least one tracking system comprises the inclusion of at least one amine associated with or included in the delivery vehicle. [0731] In some embodiments, the at least one tracking system comprises the inclusion of at least one Texas Red® associated with or included in the delivery vehicle. [0732] In some embodiments, the at least one tracking system comprises the inclusion of at least one biotin associated with, encoded in or included in the benchmark construct. [0733] In some embodiments, the at least one tracking system comprises the inclusion of at least one digoxygenin associated with, encoded in or included in the benchmark construct. [0734] In some embodiments, the at least one tracking system comprises the inclusion of at least one dinitrophenyl (DNP) associated with, encoded in or included in the benchmark construct. [0735] In some embodiments, the at least one tracking system comprises the inclusion of at least one Fluorescein associated with, encoded in or included in the benchmark construct. [0736] In some embodiments, the at least one tracking system comprises the inclusion of at least one fucose associated with, encoded in or included in the benchmark construct. [0737] In some embodiments, the at least one tracking system comprises the inclusion of at least one amine associated with, encoded in or included in the benchmark construct. [0738] In some embodiments, the at least one tracking system comprises the inclusion of at least one Texas Red® associated with, encoded in or included in the benchmark construct. [0739] In some embodiments, the at least one tracking system comprises the inclusion of at least one reporter sequence or protein associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one reporter sequence or protein associated with, encoded in or included in the benchmark construct. Non-limiting examples of reporter sequence or protein include eGFP, luciferase, gene editor (e.g. cas9 edit, DNA readout), ox-40, beta6 integrin, CD45, a surface marker with a HA tag, flag tag with or without a TEV protease site, or any combination thereof. [0740] In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein associated with, encoded in or included in the benchmark construct. Non-limiting examples of functional sequence or protein include fluorescent protein, a surface protein, Cre-Recombinase, CRISPR/CAS system, surface protein with an epitope tag (e.g., HA, FLAG, etc.) or any combination thereof [0741] In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein that comprises a protease cleavage site (e.g., TEV) which may be associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein that comprises a protease cleavage site (e.g., TEV) which may be associated with, encoded in or included in the benchmark construct. [0742] In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein that comprises an affinity tag (e.g.3xHA, FLAG, His) which may be associated with or included in the delivery vehicle. In some embodiments, the at least one tracking system comprises the inclusion of at least one functional sequence or protein that comprises an affinity tag (e.g.3xHA, FLAG, His) which may be associated with, encoded in or included in the benchmark construct. V. PHARMACEUTICAL COMPOSITION AND ROUTE OF ADMINISTRATION Pharmaceutical Compositions and Formulations [0743] The originator constructs, benchmark constructs, and targeting systems can be formulated using one or more excipients to: (1) increase stability; (2) increase cell transfection or transduction; (3) permit the sustained or delayed expression of the payload; (4) alter the biodistribution (e.g., target the viral particle to specific tissues or cell types); (5) increase the translation of encoded protein; (6) alter the release profile of encoded protein; and/or (7) allow for regulatable expression of the cargo and/or payload. [0744] Formulations can include, without limitation, saline, liposomes, lipid nanoparticles, polymers, peptides, proteins, cells transfected with viral vectors (e.g., for transfer or transplantation into a subject) and combinations thereof. [0745] Formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. As used herein the term "pharmaceutical composition" refers to compositions comprising at least one active ingredient and optionally one or more pharmaceutically acceptable excipients. [0746] In general, such preparatory methods include the step of associating the active ingredient with an excipient and/or one or more other accessory ingredients. As used herein, the phrase "active ingredient" generally refers either to an originator construct or benchmark construct with a payload region or cargo or payload as described herein. [0747] Formulations of the originator constructs, benchmark constructs, and targeting systems and pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with an excipient and/or one or more other accessory ingredients, and then, if necessary and/or desirable, dividing, shaping and/or packaging the product into a desired single- or multi-dose unit. [0748] A pharmaceutical composition in accordance with the present disclosure may be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a "unit dose" refers to a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage. [0749] In some embodiments, a pharmaceutically acceptable excipient may be at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, an excipient is approved for use for humans and for veterinary use. In some embodiments, an excipient may be approved by United States Food and Drug Administration. In some embodiments, an excipient may be of pharmaceutical grade. In some embodiments, an excipient may meet the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the International Pharmacopoeia. [0750] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary, depending upon the identity, size, and/or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may comprise between 0.1% and 99% (w/w) of the active ingredient. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, or at least 80% (w/w) active ingredient. [0751] In one aspect, the present disclosure further provides delivery systems for delivery of a therapeutic payload disclosed herein. In some embodiments, a delivery system suitable for delivery of the therapeutic payload disclosed herein comprises a lipid nanoparticle (LNP) formulation. [0752] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a phospholipid. In alternative embodiments, an LNP comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a zwitterionic amino acid lipid. In some embodiments, an LNP further comprises a 5 th lipid, besides any of the aforementioned lipid components. In some embodiments, the LNP encapsulates one or more elements of the active agent of the present disclosure. In some embodiments, an LNP further comprises a targeting moiety covalently or non-covalently bound to the outer surface of the LNP. In some embodiments, the targeting moiety is a targeting moiety that binds to, or otherwise facilitates uptake by, cells of a particular organ system. [0753] In some embodiments, an LNP has a diameter of at least about 20nm, 30 nm, 40nm, 50nm, 60nm, 70nm, 80nm, or 90nm. In some embodiments, an LNP has a diameter of less than about 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, or 160nm. In some embodiments, an LNP has a diameter of less than about 100nm. In some embodiments, an LNP has a diameter of less than about 90nm. In some embodiments, an LNP has a diameter of less than about 80nm. In some embodiments, an LNP has a diameter of about 60-100nm. In some embodiments, an LNP has a diameter of about 75-80nm. [0754] In some embodiments, the lipid nanoparticle compositions of the present disclosure are described according to the respective molar ratios of the component lipids in the formulation. As a non-limiting example, the mol-% of the ionizable lipid may be from about 10 mol-% to about 80 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 20 mol-% to about 70 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 30 mol-% to about 60 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 35 mol-% to about 55 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 40 mol-% to about 50 mol-%. [0755] In some embodiments, the mol-% of the phospholipid may be from about 1 mol-% to about 50 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 2 mol-% to about 45 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 3 mol-% to about 40 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 4 mol-% to about 35 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 5 mol-% to about 30 mol-%. In some embodiments, the mol- % of the phospholipid may be from about 10 mol-% to about 20 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 5 mol-% to about 20 mol-%. [0756] In some embodiments, the mol-% of the structural lipid may be from about 10 mol-% to about 80 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 20 mol-% to about 70 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 30 mol-% to about 60 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 35 mol-% to about 55 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 40 mol-% to about 50 mol-%. [0757] In some embodiments, the mol-% of the PEG lipid may be from about 0.1 mol-% to about 10 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 0.2 mol-% to about 5 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 0.5 mol-% to about 3 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 1 mol-% to about 2 mol-%. In some embodiments, the mol-% of the PEG lipid may be about 1.5 mol-%. [0758] In some embodiments, a nanoparticle includes an ionizable lipid, a phospholipid, a PEG lipid, and a structural lipid. In certain embodiments, the lipid component of the nanoparticle composition includes about 30 mol % to about 60 mol % ionizable lipid, about 0 mol % to about 30 mol % phospholipid, about 18.5 mol % to about 48.5 mol % structural lipid, and about 0 mol% to about 10 mol% of PEG lipid, provided that the total mol % does not exceed 100%. In some embodiments, the lipid component of the nanoparticle composition includes about 35 mol % to about 55 mol % ionizable lipid, about 5 mol % to about 25 mol % phospholipid, about 30 mol % to about 40 mol % structural lipid, and about 0 mol % to about 10 mol % of PEG lipid. In a particular embodiment, the lipid component includes about 50 mol % ionizable lipid, about 10 mol % phospholipid, about 38.5 mol % structural lipid, and about 1.5 mol% of PEG lipid. In another particular embodiment, the lipid component includes about 40 mol % ionizable lipid, about 20 mol % phospholipid, about 38.5 mol % structural lipid, and about 1.5 mol % of PEG lipid. In another particular embodiment, the lipid component includes about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 40 mol % structural lipid, and about 1.5 mol % of PEG lipid. In another particular embodiment, the lipid component includes about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 39 mol % structural lipid, and about 2.5 mol % of PEG lipid. In some embodiments, the phospholipid may be DOPE or DSPC. In other embodiments, the PEG lipid may be PEG-DMG and/or the structural lipid may be cholesterol. The amount of active agent in a nanoparticle composition may depend on the size, composition, desired target and/or application, or other properties of the nanoparticle composition as well as on the properties of the active agent. For example, the amount of active agent useful in a nanoparticle composition may depend on the size, sequence, and other characteristics of the active agent. The relative amounts of active agent and other elements (e.g., lipids) in a nanoparticle composition may also vary. In some embodiments, the wt/wt ratio of the lipid component to an enzyme in a nanoparticle composition may be from about 5:1 to about 60:1, such as 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, and 60:1. The amount of a enzyme in a nanoparticle composition may, for example, be measured using absorption spectroscopy (e.g., ultraviolet-visible spectroscopy). [0759] In some embodiments, a nanoparticle composition comprising an active agent of the present disclosure is formulated to provide a specific E:P ratio. The E:P ratio of the composition refers to the molar ratio of nitrogen atoms in one or more lipids to the number of phosphate groups in an RNA active agent. In general, a lower E:P ratio is preferred. The one or more enzymes, lipids, and amounts thereof may be selected to provide an E:P ratio from about 2:1 to about 30:1, such as 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, or 30:1. In certain embodiments, the E:P ratio may be from about 2:1 to about 8:1. In other embodiments, the E:P ratio is from about 5:1 to about 8:1. For example, the E:P ratio may be about 5.0:1, about 5.5:1, about 5.67:1, about 6.0:1, about 6.5:1, or about 7.0:1. [0760] The characteristics of a nanoparticle composition may depend on the components thereof. For example, a nanoparticle composition including cholesterol as a structural lipid may have different characteristics than a nanoparticle composition that includes a different structural lipid. Similarly, the characteristics of a nanoparticle composition may depend on the absolute or relative amounts of its components. For instance, a nanoparticle composition including a higher molar fraction of a phospholipid may have different characteristics than a nanoparticle composi tion including a lower molar fraction of a phospholipid. Characteristics may also vary depending on the method and conditions of preparation of the nanoparticle composition. Nanoparticle compositions may be characterized by a variety of methods. For example, microscopy (e.g., transmission electron microscopy or scanning electron microscopy) may be used to examine the morphology and size distribution of a nanoparticle composition. Dynamic light scattering or potentiometry (e.g., potentiometric titrations) may be used to measure Zeta potentials. Dynamic light scattering may also be utilized to determine particle sizes. Instruments such as the Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, Worcestershire, UK) may also be used to measure multiple characteristics of a nanoparticle composition, Such as particle size, polydispersity index, and Zeta potential. [0761] The mean size of a nanoparticle composition may be between 10s of nm and 100s of nm, e.g., measured by dynamic light scattering (DLS). For example, the mean size may be from about 40 nm to about 150 nm, such as about 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm. In some embodiments, the mean size of a nanoparticle composition may be from about 50 nm to about 100 nm, from about 50 nm to about 90 nm, from about 50 nm to about 80 nm, from about 50 nm to about 70 nm, from about 50 nm to about 60 nm, from about 60 nm to about 100 nm, from about 60 nm to about 90 nm, from about 60 nm to about 80 nm, from about 60 nm to about 70 nm, from about 70 nm to about 100 nm, from about 70 nm to about 90 nm, from about 70 nm to about 80 nm, from about 80 nm to about 100 nm, from about 80 nm to about 90 nm, or from about 90 nm to about 100 nm. In certain embodiments, the mean size of a nanoparticle composition may be from about 70 nm to about 100 nm. In a particular embodiment, the mean size may be about 80 nm. In other embodiments, the mean size may be about 100 nm. [0762] A nanoparticle composition may be relatively homogenous. A polydispersity index may be used to indicate the homogeneity of a nanoparticle composition, e.g., the particle size distribution of the nanoparticle compositions. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. A nanoparticle composition may have a polydispersity index from about 0 to about 0.25, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, or 0.25. [0763] The Zeta potential of a nanoparticle composition may be used to indicate the electrokinetic potential of the composition. For example, the Zeta potential may describe the surface charge of a nanoparticle composition. Nanoparticle compositions with relatively low charges, positive or negative, are generally desirable, as more highly charged species may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the Zeta potential of a nanoparticle composition may be from about -10 mV to about +20 mV, from about -10 mV to about +15 mV, from about -10 mV to about +10 mV, from about -10 mV to about +5 mV, from about -10 mV to about 0 mV, from about -10 mV to about -5 mV, from about -5 mV to about +20 mV, from about -5 mV to about +15 mV, from about -5 mV to about +10 mV, from about -5 mV to about +5 mV, from about -5 mV to about 0 mV, from about 0 mV, to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV to about +10 mV, from about 0 mV to about +5 mV, from about +5 mV to about +20 mV, from about +5 mV, to about +15 mV, or from about +5 mV to about +10 mV. [0764] The efficiency of encapsulation of a payload describes the amount of payload that is encapsulated or otherwise associated with a nanoparticle composition after preparation, relative to the initial amount provided. The encapsulation efficiency is desirably high (e.g., close to 100%). The encapsulation efficiency may be measured, for example, by comparing the amount of payload in a solution containing the nanoparticle composition before and after breaking up the nanoparticle composition with one or more organic solvents or detergents. Fluorescence may be used to measure the amount of free payload in a solution. For the nanoparticle compositions described herein, the encapsulation efficiency of a therapeutic and/or prophylactic may be at least 50%, for example 50%, 55%, 60%.65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the encapsulation efficiency may be at least 80%. In certain embodiments, the encapsulation efficiency may be at least 90%. [0765] Lipids and their method of preparation are disclosed in, e.g., U.S. Patent Nos. 8,569,256, 5,965,542 and U.S. Patent Publication Nos. 2016/0199485, 2016/0009637, 2015/0273068, 2015/0265708, 2015/0203446, 2015/0005363, 2014/0308304, 2014/0200257, 2013/086373, 2013/0338210, 2013/0323269, 2013/0245107, 2013/0195920, 2013/0123338, 2013/0022649, 2013/0017223, 2012/0295832, 2012/0183581, 2012/0172411, 2012/0027803, 2012/0058188, 2011/0311583, 2011/0311582, 2011/0262527, 2011/0216622, 2011/0117125, 2011/0091525, 2011/0076335, 2011/0060032, 2010/0130588, 2007/0042031, 2006/0240093, 2006/0083780, 2006/0008910, 2005/0175682, 2005/017054, 2005/0118253, 2005/0064595, 2004/0142025, 2007/0042031, 1999/009076 and PCT Pub. Nos. WO 99/39741, WO 2017/117528, WO 2017/004143, WO 2017/075531, WO 2015/199952, WO 2014/008334, WO 2013/086373, WO 2013/086322, WO 2013/016058, WO 2013/086373, WO2011/141705, and WO 2001/07548 and Semple et. al, Nature Biotechnology, 2010, 28, 172-176, the full disclosures of which are herein incorporated by reference in their entirety for all purposes. [0766] A nanoparticle composition may include any substance useful in pharmaceutical compositions. For example, the nanoparticle composition may include one or more pharmaceutically acceptable excipients or accessory ingredients such as, but not limited to, one or more solvents, dispersion media, diluents, dispersion aids, suspension aids, granulating aids, disintegrants, fillers, glidants, liquid vehicles, binders, surface active agents, isotonic agents, thickening or emulsifying agents, buffering agents, lubricating agents, oils, preservatives, and other species. Excipients such as waxes, butters, coloring agents, coating agents, flavorings, and perfuming agents may also be included. Pharmaceutically acceptable excipients are well known in the art (see for example Remington’s The Science and Practice of Pharmacy, 21 st Edition, A. R. Gennaro: Lippincott, Williams & Wilkins, Baltimore, Md., 2006). Excipients and Diluents [0767] Excipients, as used herein, include, but are not limited to, any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, and the like, as suited to the particular dosage form desired. Various excipients for formulating pharmaceutical compositions and techniques for preparing the composition are known in the art (see Remington: The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro, Lippincott, Williams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference in its entirety). The use of a conventional excipient medium may be contemplated within the scope of the present disclosure, except insofar as any conventional excipient medium may be incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition. [0768] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, etc., and/or combinations thereof. Ionizable lipids [0769] In some embodiments, an LNP disclosed herein comprises an ionizable lipid. In some embodiments, an LNP comprises two or more ionizable lipids. In some embodiments, the ionizable lipid is any ionizable lipid disclosed herein, or any combinations thereof. Structural lipids [0770] In some embodiments, an LNP comprises a structural lipid. Structural lipids can be selected from the group consisting of, but are not limited to, cholesterol, fecosterol, fucosterol, beta sitosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, cholic acid, sitostanol, litocholic acid, tomatine, ursolic acid, alpha-tocopherol, Vitamin D3, Vitamin D2, Calcipotriol, botulin, lupeol, oleanolic acid, beta-sitosterol-acetate and mixtures thereof. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid is a cholesterol analogue disclosed by Patel, et al., Nat Commun., 11, 983 (2020), which is incorporated herein by reference in its entirety. In some embodiments, the structural lipid includes cholesterol and a corticosteroid (such as prednisolone, dexamethasone, prednisone, and hydrocortisone), or any combinations thereof. In some embodiments, a structural lipid is described in international patent application WO2019152557A1, which is incorporated herein by reference in its entirety. [0771] In some embodiments, a structural lipid is a cholesterol analog. Using a cholesterol analog may enhance endosomal escape as described in Patel et al., Naturally-occuring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA, Nature Communications (2020), which is incorporated herein by reference. [0772] In some embodiments, a structural lipid is a phytosterol. Using a phytosterol may enhance endosomal escape as described in Herrera et al., Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery, Biomaterials Science (2020), which is incorporated herein by reference. [0773] In some embodiments, a structural lipid contains plant sterol mimetics for enhanced endosomal release. PEGylated lipids [0774] A PEGylated lipid is a lipid modified with polyethylene glycol. In some embodiments, the LNP comprises a compound of Formula I or a pharmaceutically acceptable salt thereof, as described herein above. In some embodiments, the LNP comprises a compound of Formula II or a pharmaceutically acceptable salt thereof, as described herein above. In some embodiments, the LNP comprises a compound set forth in Table (I), as described herein above. [0775] In some embodiments, an LNP comprises an additional PEGylated lipid or PEG- modified lipid. A PEGylated lipid may be selected from the non-limiting group consisting of PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and mixtures thereof. For example, a PEG lipid may be PEG-c-DOMG, PEG- DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or a PEG-DSPE lipid. [0776] In some embodiments, the LNP comprises a PEGylated lipid disclosed in one of US 2019/0240354; US 2010/0130588; US 2021/0087135; WO 2021/204179; US 2021/0128488; US 2020/0121809; US 2017/0119904; US 2013/0108685; US 2013/0195920; US 2015/0005363; US 2014/0308304; US 2013/0053572; WO 2019/232095A1; WO 2021/077067; WO 2019/152557; US 2015/0203446; US 2017/0210697; US 2014/0200257; or WO 2019/089828A1, each of which is incorporated by reference herein in their entirety. [0777] In some embodiments, the LNP comprises a PEGylated lipid substitute in place of the PEGylated lipid. All embodiments disclosed herein that contemplate a PEGylated lipid should be understood to also apply to PEGylated lipid substitutes. In some embodiments, the LNP comprises a polysarcosine-lipid conjugate, such as those disclosed in US 2022/0001025 A1, which is incorporated by reference herein in its entirety. Phospholipids [0778] In some embodiments, an LNP of the present disclosure comprises a phospholipid. Phospholipids useful in the compositions and methods may be selected from the non-limiting group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn- glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1.2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2- diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3- phosphocho line (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuc cinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1- hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3- phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl- sn-glycero-3-phosphocholine, 1,2-diphytanoylsn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3- phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2- diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3- phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sodium (S)-2-ammonio-3-((((R)-2-(oleoyloxy)-3- (stearoyloxy)propoxy)oxidophosphoryl)oxy)propanoate (L-α-phosphatidylserine; Brain PS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleoyl-phosphatidylethanolamine4-(N- maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell-fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl-phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), 1,2-dioleoyl-sn-glycero-3-phosphate (18:1 PA; DOPA), ammonium bis((S)-2-hydroxy- 3-(oleoyloxy)propyl) phosphate (18:1 DMP; LBPA), 1,2-dioleoyl-sn-glycero-3-phospho-(1’- myo-inositol) (DOPI; 18:1 PI), 1,2-distearoyl-sn-glycero-3-phospho-L-serine (18:0 PS), 1,2- dilinoleoyl-sn-glycero-3-phospho-L-serine (18:2 PS), 1-palmitoyl-2-oleoyl-sn-glycero-3- phospho-L-serine (16:0-18:1 PS; POPS), 1-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (18:0-18:1 PS), 1-stearoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine (18:0-18:2 PS), 1- oleoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:1 Lyso PS), 1-stearoyl-2-hydroxy-sn- glycero-3-phospho-L-serine (18:0 Lyso PS), and sphingomyelin. In some embodiments, an LNP includes DSPC. In certain embodiments, an LNP includes DOPE. In some embodiments, an LNP includes both DSPC and DOPE. [0779] In some embodiments, a phospholipid tail may be modified in order to promote endosomal escape as described in U.S. 2021/0121411, which is incorporated herein by reference. [0780] In some embodiments, the LNP comprises a phospholipid disclosed in one of US 2019/0240354; US 2010/0130588; US 2021/0087135; WO 2021/204179; US 2021/0128488; US 2020/0121809; US 2017/0119904; US 2013/0108685; US 2013/0195920; US 2015/0005363; US 2014/0308304; US 2013/0053572; WO 2019/232095A1; WO 2021/077067; WO 2019/152557; US 2017/0210697; or WO 2019/089828A1, each of which is incorporated by reference herein in their entirety. [0781] In some embodiments, phospholipids disclosed in US 2020/0121809 have the following structure: [0782] wherein R1 and R2 are each independently a branched or straight, saturated or unsaturated carbon chain (e.g., alkyl, alkenyl, alkynyl). Targeting moieties [0783] In some embodiments, the lipid nanoparticle further comprises a targeting moiety. The targeting moiety may be an antibody or a fragment thereof. The targeting moiety may be capable of binding to a target antigen. [0784] In some embodiments, the pharmaceutical composition comprises a targeting moiety that is operably connected to a lipid nanoparticle. In some embodiments, the targeting moiety is capable of binding to a target antigen. In some embodiments, the target antigen is expressed in a target organ. In some embodiments, the target antigen is expressed more in the target organ than it is in the liver. [0785] In some embodiments, the targeting moiety is an antibody as described in WO2016189532A1, which is incorporated herein by reference. For example, in some embodiments, the targeted particles are conjugated to a specific anti-CD38 monoclonal antibody (mAb), which allows specific delivery of the siRNAs encapsulated within the particles at a greater percentage to B-cell lymphocytes malignancies (such as MCL) than to other subtypes of leukocytes. [0786] In some embodiments, the lipid nanoparticles may be targeted when conjugated/attached/associated with a targeting moiety such as an antibody. Zwitterionic amino lipids [0787] In some embodiments, an LNP comprises a zwitterionic lipid. In some embodiments, an LNP comprising a zwitterionic lipid does not comprise a phospholipid. [0788] Zwitterionic amino lipids have been shown to be able to self-assemble into LNPs without phospholipids to load, stabilize, and release mRNAs intracellular as described in U.S. Patent Application 20210121411, which is incorporated herein by reference in its entirety. Zwitterionic, ionizable cationic and permanently cationic helper lipids enable tissue-selective mRNA delivery and CRISPR-Cas9 gene editing in spleen, liver and lungs as described in Liu et al., Membrane-destablizing ionizable phospholipids for organ-selective mRNA delivery and CRISPR-Cas gene editing, Nat Mater. (2021), which is incorporated herein by reference in its entirety. [0789] The zwitterionic lipids may have head groups containing a cationic amine and an anionic carboxylate as described in Walsh et al., Synthesis, Characterization and Evaluation of Ionizable Lysine-Based Lipids for siRNA Delivery, Bioconjug Chem. (2013), which is incorporated herein by reference in its entirety. Ionizable lysine-based lipids containing a lysine head group linked to a long-chain dialkylamine through an amide linkage at the lysine α-amine may reduce immunogenicity as described in Walsh et al., Synthesis, Characterization and Evaluation of Ionizable Lysine-Based Lipids for siRNA Delivery, Bioconjug Chem. (2013). Additional Lipid s [0790] In some embodiments, the LNP compositions of the present disclosure further comprise one or more additional lipid components capable of influencing the tropism of the LNP. In some embodiments, the LNP further comprises at least one lipid selected from DDAB, EPC, 14PA, 18BMP, DODAP, DOTAP, and C12-200 (see Cheng, et al. Nat Nanotechnol. 2020 April; 15(4): 313–320.; Dillard, et al. PNAS 2021 Vol.118 No.52.). Polynucleotides [0791] In some embodiments, an LNP of the present disclosure contains an active agent. In some embodiments, an active agent is a polynucleotide. In some embodiments, a LNP is capable of delivering a polynucleotide to a target organ. A polynucleotide, in its broadest sense of the term, includes any compound and/or substance that is or can be incorporated into an oligonucleotide chain. Exemplary polynucleotides for use in accordance with the present disclosure include, but are not limited to, one or more of deoxyribonucleic acid (DNA), ribonucleic acid (RNA) including messenger mRNA (mRNA), hybrids thereof, RNAi- inducing agents, RNAi agents, siRNAs, shRNAs, miRNAs, antisense RNAs, ribozymes, catalytic DNA, RNAs that induce triple helix formation, aptamers, vectors, etc. RNAs useful in the compositions and methods described herein can be selected from the group consisting of but are not limited to, shortimers, antagomirs, antisense, ribozymes, short interfering RNA (siRNA), asymmetrical interfering RNA (aiRNA), microRNA (miRNA), Dicer substrate RNA (dsRNA), short hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and mixtures thereof. In some embodiments, a polynucleotide is mRNA. In some embodiments, a polynucleotide is circular RNA. In some embodiments, a polynucleotide encodes a protein. A polynucleotide may encode any polypeptide of interest, including any naturally or non- naturally occurring or otherwise modified polypeptide. A polypeptide may be of any size and may have any secondary structure or activity. In some embodiments, a polypeptide encoded by an mRNA may have a therapeutic effect when expressed in a cell. [0792] In other embodiments, a polynucleotide is an siRNA. An siRNA may be capable of selectively knocking down or down regulating expression of a gene of interest. For example, an siRNA could be selected to silence a gene associated with a particular disease, disorder, or condition upon administration to a subject in need thereof of a nanoparticle composition including the siRNA. An siRNA may comprise a sequence that is complementary to an mRNA sequence that encodes a gene or protein of interest. In some embodiments, the siRNA may be an immunomodulatory siRNA. [0793] In some embodiments, a polynucleotide is an shRNA or a vector or plasmid encoding the same. An shRNA may be produced inside a target cell upon delivery of an appropriate construct to the nucleus. Constructs and mechanisms relating to shRNA are well known in the relevant arts. [0794] A polynucleotide may include a first region of linked nucleosides encoding a polypeptide of interest (e.g., a coding region), a first flanking region located at the 5'-terminus of the first region (e.g., a 5'-UTR), a second flanking region located at the 3'-terminus of the first region (e.g., a 3'-UTR), at least one 5'-cap region, and a 3'-stabilizing region. In some embodiments, a polynucleotide further includes a poly-A region or a Kozak sequence (e.g., in the 5'-UTR). In some cases, polynucleotides may contain one or more intronic nucleotide sequences capable of being excised from the polynucleotide. In some embodiments, a polynucleotide (e.g., an mRNA) may include a 5'cap structure, a chain terminating nucleotide, a stem loop, a polyA sequence, and/or a polyadenylation signal. Any one of the regions of a nucleic acid may include one or more alternative components (e.g., an alternative nucleoside). For example, the 3'-stabilizing region may contain an alternative nucleoside such as an L- nucleoside, an inverted thymidine, or a 2'-O-methyl nucleoside and/or the coding region, 5'- UTR, 3'-UTR, or cap region may include an alternative nucleoside such as a 5-substituted uridine (e.g., 5-methoxyu ridine), a 1-substituted pseudouridine (e.g., 1-methyl pseudouridine or 1-ethyl-pseudouridine), and/or a 5-substituted cytidine (e.g., 5-methyl-cytidine). In some embodiments, a polynucleotide contains only naturally occurring nucleosides. [0795] In some cases, a polynucleotide is greater than 30 nucleotides in length. In another embodiment, the poly nucleotide molecule is greater than 35 nucleotides in length. In another embodiment, the length is at least 40 nucleotides. In another embodiment, the length is at least 45 nucleotides. In another embodiment, the length is at least 55 nucleotides. In another embodiment, the length is at least 50 nucleotides. In another embodiment, the length is at least 60 nucleotides. In another embodiment, the length is at least 80 nucleotides. In another embodiment, the length is at least 90 nucleotides. In another embodiment, the length is at least 100 nucleotides. In another embodiment, the length is at least 120 nucleotides. In another embodiment, the length is at least 140 nucleotides. In another embodiment, the length is at least 160 nucleotides. In another embodiment, the length is at least 180 nucleotides. In another embodiment, the length is at least 200 nucleotides. In another embodiment, the length is at least 250 nucleotides. In another embodiment, the length is at least 300 nucleotides. In another embodiment, the length is at least 350 nucleotides. In another embodiment, the length is at least 400 nucleotides. In another embodiment, the length is at least 450 nucleotides. In another embodiment, the length is at least 500 nucleotides. In another embodiment, the length is at least 600 nucleotides. In another embodiment, the length is at least 700 nucleotides. In another embodiment, the length is at least 800 nucleotides. In another embodiment, the length is at least 900 nucleotides. In another embodiment, the length is at least 1000 nucleotides. In another embodiment, the length is at least 1100 nucleotides. In another embodiment, the length is at least 1200 nucleotides. In another embodiment, the length is at least 1300 nucleotides. In another embodiment, the length is at least 1400 nucleotides. In another embodiment, the length is at least 1500 nucleotides. In another embodiment, the length is at least 1600 nucleotides. In another embodiment, the length is at least 1800 nucleotides. In another embodiment, the length is at least 2000 nucleotides. In another embodiment, the length is at least 2500 nucleotides. In another embodiment, the length is at least 3000 nucleotides. In another embodiment, the length is at least 4000 nucleotides. In another embodiment, the length is at least 5000 nucleotides, or greater than 5000 nucleotides. [0796] In some embodiments, a polynucleotide molecule, formula, composition or method associated therewith comprises one or more polynucleotides comprising features as described in WO2002/098443, WO2003/051401, WO2008/052770, WO2009/127230, WO2006/122828, WO2008/083949, WO2010/088927, WO2010/037539, WO2004/004743, WO2005/016376, WO2006/024518, WO2007/095,976, WO2008/014979, WO2008/077592, WO2009/030481, WO2009/095226, WO2011/069586, WO2011/026641, WO2011/144358, WO2012/019780, WO2012/013326, WO2012/089338, WO2012/113513, WO2012/116811, WO2012/116810, WO2013/113502, WO2013/113501, WO2013/113736, WO2013/143698, WO2013/143699, WO2013/143700, WO2013/120626, WO2013/120627, WO2013/120628, WO2013/120629, WO2013/174409, WO2014/127917, WO2015/024669, WO2015/024668, WO2015/024667, WO2015/024665, WO2015/024666, WO2015/024664, WO2015/101415, WO2015/101414, WO2015/024667, WO2015/062738, WO2015/101416, all of which are incorporated by reference herein. [0797] Polynucleotides, such as circular RNA, may contain an internal ribosome entry site (IRES). An IRES may act as the sole ribosome binding site, or may serve as one of multiple ribosome binding sites of an mRNA. A polynucleotide containing more than one functional ribosome binding site may encode several peptides or polypeptides that are translated independently by the ribosomes (e.g., multicistronic mRNA). When polynucleotides are provided with an IRES, further optionally provided is a second translatable region. Examples of IRES sequences that can be used according to the present disclosure include without limitation, those from picornaviruses (e.g., FMDV), pest viruses (CFFV), polio viruses (PV), encephalomyocarditis viruses (ECMV), foot-and mouth disease viruses (FMDV), hepatitis C viruses (HCV), classical Swine fever viruses (CSFV), murine leukemia virus (MLV), simian immune deficiency viruses (SIV) or cricket paralysis viruses (CrPV). [0798] In some embodiments, a polynucleotide comprises one or more microRNA binding sites. In some embodiments, a microRNA binding site is recognized by a microRNA in a non- target organ. In some embodiments, a microRNA binding site is recognized by a microRNA in the liver. In some embodiments, a microRNA binding site is recognized by a microRNA in hepatic cells. Inactive Ingredients [0799] In some embodiments, formulations described herein may comprise at least one inactive ingredient. As used herein, the term "inactive ingredient" refers to one or more agents that do not contribute to the activity of the active ingredient of the pharmaceutical composition included in formulations. In some embodiments, all, none or some of the inactive ingredients which may be used in the formulations of the present disclosure may be approved by the US Food and Drug Administration (FDA). [0800] In some embodiments, formulations disclosed herein may include cations or anions. The formulations include metal cations such as, but not limited to, Zn 2+ , Ca 2+ , Cu 2+ , Mn 2+ , Mg 2+ , and combinations thereof. As a non-limiting example, formulations may include polymers and complexes with a metal cation. [0801] Formulations of the disclosure may also include one or more pharmaceutically acceptable salts. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form (e.g., by reacting the free base group with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. Representative acid addition salts include acetate, acetic acid, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzene sulfonic acid, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. The pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. [0802] Solvates may be prepared by crystallization, recrystallization, or precipitation from a solution that includes organic solvents, water, or a mixture thereof. Examples of suitable solvents are ethanol, water (for example, mono-, di-, and tri-hydrates), N-methylpyrrolidinone (NMP), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF), N,N'- dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU), 1,3-dimethyl-3,4,5,6- tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone, benzyl benzoate, and the like. When water is the solvent, the solvate is referred to as a "hydrate." Routes of Administration [0803] The originator constructs, benchmark constructs, and targeting systems described herein may be administered by any delivery route which results in a therapeutically effective outcome. These include, but are not limited to, enteral (into the intestine), gastroenteral, epidural (into the dura mater), oral (by way of the mouth), transdermal, intracerebral (into the cerebrum), intracerebroventricular (into the cerebral ventricles), epicutaneous (application onto the skin), intradermal (into the skin itself), subcutaneous (under the skin), nasal administration (through the nose), intravenous (into a vein), intravenous bolus, intravenous drip, intra-arterial (into an artery), intramuscular (into a muscle), intracardiac (into the heart), intraosseous infusion (into the bone marrow), intrathecal (into the spinal canal), intraparenchymal (into brain tissue), intraperitoneal (infusion or injection into the peritoneum), intravesical infusion, intravitreal (through the eye), intracavernous injection (into a pathologic cavity) intracavitary (into the base of the penis), intravaginal administration, intrauterine, extra-amniotic administration, transdermal (diffusion through the intact skin for systemic distribution), transmucosal (diffusion through a mucous membrane), transvaginal, insufflation (snorting), sublingual, sublabial, enema, eye drops (onto the conjunctiva), ear drops, auricular (in or by way of the ear), buccal (directed toward the cheek), conjunctival, cutaneous, dental (to a tooth or teeth), electro-osmosis, endocervical, endosinusial, endotracheal, extracorporeal, hemodialysis, infiltration, interstitial, intra-abdominal, intra-amniotic, intra-articular, intrabiliary, intrabronchial, intrabursal, intracartilaginous (within a cartilage), intracaudal (within the cauda equine), intracisternal (within the cisterna magna cerebellomedularis), intracorneal (within the cornea), dental intracoronal, intracoronary (within the coronary arteries), intracorporus cavernosum (within the dilatable spaces of the corporus cavernosa of the penis), intradiscal (within a disc), intraductal (within a duct of a gland), intraduodenal (within the duodenum), intradural (within or beneath the dura), intraepidermal (to the epidermis), intraesophageal (to the esophagus), intragastric (within the stomach), intragingival (within the gingivae), intraileal (within the distal portion of the small intestine), intralesional (within or introduced directly to a localized lesion), intraluminal (within a lumen of a tube), intralymphatic (within the lymph), intramedullary (within the marrow cavity of a bone), intrameningeal (within the meninges), intramyocardial (within the myocardium), intraocular (within the eye), intraovarian (within the ovary), intrapericardial (within the pericardium), intrapleural (within the pleura), intraprostatic (within the prostate gland), intrapulmonary (within the lungs or its bronchi), intrasinal (within the nasal or periorbital sinuses), intraspinal (within the vertebral column), intrasynovial (within the synovial cavity of a joint), intratendinous (within a tendon), intratesticular (within the testicle), intrathecal (within the cerebrospinal fluid at any level of the cerebrospinal axis), intrathoracic (within the thorax), intratubular (within the tubules of an organ), intratumor (within a tumor), intratympanic (within the aurus media), intravascular (within a vessel or vessels), intraventricular (within a ventricle), iontophoresis (by means of electric current where ions of soluble salts migrate into the tissues of the body), irrigation (to bathe or flush open wounds or body cavities), laryngeal (directly upon the larynx), nasogastric (through the nose and into the stomach), occlusive dressing technique (topical route administration which is then covered by a dressing which occludes the area), ophthalmic (to the external eye), oropharyngeal (directly to the mouth and pharynx), parenteral, percutaneous, periarticular, peridural, perineural, periodontal, rectal, respiratory (within the respiratory tract by inhaling orally or nasally for local or systemic effect), retrobulbar (behind the pons or behind the eyeball), soft tissue, subarachnoid, subconjunctival, submucosal, topical, transplacental (through or across the placenta), transtracheal (through the wall of the trachea), transtympanic (across or through the tympanic cavity), ureteral (to the ureter), urethral (to the urethra), vaginal, caudal block, diagnostic, nerve block, biliary perfusion, cardiac perfusion, photopheresis, and spinal. [0804] In some embodiments, compositions may be administered in a way which allows them to cross the blood-brain barrier, vascular barrier, or other epithelial barrier. The originator constructs, benchmark constructs, and targeting systems may be administered in any suitable form, either as a liquid solution or suspension, as a solid form suitable for liquid solution or suspension in a liquid solution. The originator constructs, benchmark constructs, and targeting systems may be formulated with any appropriate and pharmaceutically acceptable excipient. [0805] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered to a subject via a single route administration. [0806] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered to a subject via a multi-site route of administration. A subject may be administered at 2, 3, 4, 5, or more than 5 sites. [0807] In some embodiments, a subject may be administered the originator constructs, benchmark constructs, and targeting systems using a bolus infusion. [0808] In some embodiments, a subject may be administered originator constructs, benchmark constructs, and targeting systems using sustained delivery over a period of minutes, hours, or days. The infusion rate may be changed depending on the subject, distribution, formulation or another delivery parameter. [0809] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by intramuscular delivery route. Non-limiting examples of intramuscular administration include an intravenous injection or a subcutaneous injection. [0810] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by oral administration. Non-limiting examples of oral delivery include a digestive tract administration and a buccal administration. [0811] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by intraocular delivery route. A non-limiting example of intraocular delivery include an intravitreal injection. [0812] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by intranasal delivery route. Non-limiting examples of intranasal delivery include nasal drops or nasal sprays. [0813] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by peripheral injections. Non-limiting examples of peripheral injections include intraperitoneal, intramuscular, intravenous, conjunctival, or joint injection. [0814] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by injection into the cerebrospinal fluid. Non-limiting examples of delivery to the cerebrospinal fluid include intrathecal and intracerebroventricular administration. [0815] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by systemic delivery. As a non-limiting example, the systemic delivery may be by intravascular administration. [0816] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by intracranial delivery. [0817] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by intraparenchymal administration. [0818] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by intramuscular administration. [0819] In some embodiments, the originator constructs, benchmark constructs, and targeting systems are administered to a subject and transduce muscle of a subject. As a non-limiting example, the originator constructs, benchmark constructs, and targeting systems are administered by intramuscular administration. [0820] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by intravenous administration. [0821] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by subcutaneous administration. [0822] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be administered to a subject by topical administration. [0823] In some embodiments, the originator constructs, benchmark constructs, and targeting systems may be delivered by more than one route of administration. [0824] The originator constructs, benchmark constructs, and targeting systems described herein may be co-administered in conjunction with one or more originator constructs, benchmark constructs, targeting systems, or therapeutic agents or moieties. VI. TARGET AREA, TISSUE OR CELL FOR DELIVERY [0825] The delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to specific target areas, tissues or cells using the methods and targeted delivery systems described herein. Tumors [0826] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to a tumor. The tumor may be a benign tumor or a malignant tumor. [0827] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is localized to a connective tissue tumor such as, but not limited to, adult fibrous tissue, embryonic (myxomatous) fibrous tissue, fat tissue, cartilage, bone, and notochord. As a non-limiting example, the tumor is a benign tumor called fibroma located in adult fibrous tissue. As a non-limiting example, the tumor is a malignant tumor called fibrosarcoma located in adult fibrous tissue. As a non-limiting example, the tumor is a benign tumor called myxoma located in embryonic fibrous tissue. As a non-limiting example, the tumor is a malignant tumor called myxosarcoma located in embryonic fibrous tissue. As a non- limiting example, the tumor is a benign tumor called lipoma located in fat tissue. As a non- limiting example, the tumor is a malignant tumor called liposarcoma located in fat tissue. As a non-limiting example, the tumor is a benign tumor called chondroma located in cartilage. As a non-limiting example, the tumor is a malignant tumor called chondrosarcoma located in cartilage. As a non-limiting example, the tumor is a benign tumor called osteoma located in bone. As a non-limiting example, the tumor is a malignant tumor called osteosarcoma located in bone. As a non-limiting example, the tumor is a malignant tumor called chordoma located in notochord. As a non-limiting example, the tumor is a benign tumor called fibrous histiocytoma located in connective tissue. As a non-limiting example, the tumor is a malignant tumor called malignant fibrous histiocytoma located in connective tissue. [0828] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is localized to endothelium and/or mesothelium tumor tissue such as, but not limited to, blood vessels, lymph vessels and mesothelium. As a non-limiting example, the tumor is a benign tumor called hemangioma located in blood vessels. As a non-limiting example, the tumor is a benign tumor called hemangiopericytoma located in blood vessels. As a non-limiting example, the tumor is a malignant tumor called hemangiosarcoma located in blood vessels. As a non-limiting example, the tumor is a malignant tumor called angiosarcoma located in blood vessels. As a non-limiting example, the tumor is a benign tumor called lymphangioma located in lymph vessels. As a non-limiting example, the tumor is a malignant tumor called lymphangiosarcoma located in lymph vessels. As a non-limiting example, the tumor is a malignant tumor called mesothelioma located in the mesothelium. [0829] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is localized to blood and lymphoid cell tissue such as, but not limited to, hematopoietic cells and lymphoid tissue. As a non-limiting example, the tumor is a benign tumor called preleukemias located in hematopoietic cells. As a non-limiting example, the tumor is a benign tumor called myeloproliferative disorders located in hematopoietic cells. As a non- limiting example, the tumor is a malignant tumor called leukemia located in hematopoietic cells. As a non-limiting example, the tumor is a benign tumor called plasmacytosis located in lymphoid tissue. As a non-limiting example, the tumor a malignant tumor called plasmacytoma located in lymphoid tissue. As a non-limiting example, the tumor a malignant tumor called multiple myeloma located in lymphoid tissue. As a non-limiting example, the tumor a malignant tumor called Hodgkin lymphoma located in lymphoid tissue. As a non-limiting example, the tumor a malignant tumor called Non-Hodgkin lymphoma located in lymphoid tissue. [0830] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is localized to muscle tissue such as, but not limited to, smooth muscle and striated muscle. As a non-limiting example, the tumor is a benign tumor called Leiomyoma located in smooth muscle. As a non-limiting example, the tumor is a malignant tumor called leiomyosarcoma located in smooth muscle. As a non-limiting example, the tumor is a benign tumor called rhabdomyoma located in striated muscle. As a non-limiting example, the tumor is a malignant tumor called rhabdomyosarcoma located in striated muscle. [0831] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located to epithelial tissue such as, but not limited to, stratified squamous tissue, glandular epithelium tissue (e.g., liver, kidney, bile duct), transitional epithelium tissue, placenta and testis. As a non-limiting example, the tumor is a benign tumor called papilloma located in stratified squamous. As a non-limiting example, the tumor is a benign tumor called seborrheic keratosis located in stratified squamous. As a non-limiting example, the tumor is a malignant tumor called squamous cell carcinoma located in stratified squamous tissue. As a non-limiting example, the tumor is a malignant tumor called epidermoid carcinoma located in stratified squamous tissue. As a non-limiting example, the tumor is a benign tumor called adenoma located in glandular epithelium tissue. As a non-limiting example, the tumor is a benign tumor called hepatic adenoma located in liver glandular epithelium tissue. As a non-limiting example, the tumor is a benign tumor called renal tubular adenoma located in kidney glandular epithelium tissue. As a non-limiting example, the tumor is a benign tumor called bile duct adenoma located in bile duct glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called adenocarcinoma located in glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called hepatoma located in liver glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called hepatocellular carcinoma located in liver glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called renal cell carcinoma located in kidney glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called hypernephroma located in kidney glandular epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called cholangiocarcinoma located in bile duct glandular epithelium tissue. As a non-limiting example, the tumor is a benign tumor called transitional cell papilloma located in transitional epithelium tissue. As a non-limiting example, the tumor is a malignant tumor called transitional cell carcinoma located in transitional epithelium tissue. As a non-limiting example, the tumor is a benign tumor called hydatidiform mole located in the placenta. As a non-limiting example, the tumor is a malignant tumor called choriocarcinoma located in the placenta. As a non-limiting example, the tumor is a malignant tumor called seminoma located in the testis. As a non-limiting example, the tumor is a malignant tumor called embryonal cell carcinoma located in the testis. [0832] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located to neural tissue such as, but not limited to, glial cells, nerve cells, meninges, and nerve sheath. As a non-limiting example, the tumor is a malignant tumor called glioma (grades I-III) located in glial cells. As a non-limiting example, the tumor is a malignant tumor called anaplastic glioma (grades I-III) located in glial cells. As a non-limiting example, the tumor is a malignant tumor called glioblastoma multiforme (grade IV) located in glial cells. As a non-limiting example, the tumor is a benign tumor called ganglioneuroma located in nerve cells. As a non-limiting example, the tumor is a malignant tumor called neuroblastoma located in nerve cells. As a non-limiting example, the tumor is a malignant tumor called medulloblastoma located in nerve cells. As a non-limiting example, the tumor is a benign tumor called meningioma located in meninges tissue. As a non-limiting example, the tumor is a malignant tumor called malignant meningioma located in meninges tissue. As a non- limiting example, the tumor is a benign tumor called schwannoma located in the nerve sheath. As a non-limiting example, the tumor is a benign tumor called neurilemmoma located in the nerve sheath. As a non-limiting example, the tumor is a benign tumor called neurofibroma located in the nerve sheath. As a non-limiting example, the tumor is a malignant tumor called malignant meningioma located in the nerve sheath. As a non-limiting example, the tumor is a malignant tumor called malignant schwannoma located in the nerve sheath. As a non-limiting example, the tumor is a malignant tumor called neurofibrosarcoma located in the nerve sheath. [0833] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located to the Amine Precursor Uptake and Decarboxylation (APUD) System such as, but not limited to, pituitary tissue, parathyroid tissue, thyroid tissue, bronchial tissue, adrenalmedulla tissue, pancreas tissue, stomach and intestines, carotid body and chemo-receptor system tissue. The APUD system is a series of cells which have endocrine functions and secrete a variety of small amine or polypeptide hormones. As a non-limiting example, the tumor is a benign tumor called basophilic adenoma located in the pituitary tissue. As a non-limiting example, the tumor is a benign tumor called eosinophilic adenoma located in the pituitary tissue. As a non-limiting example, the tumor is a benign tumor called chromophobe adenoma located in the pituitary tissue. As a non-limiting example, the tumor is a benign tumor called parathyroid adenoma located in the parathyroid. As a non-limiting example, the tumor is a malignant tumor called parathyroid carcinoma located in the parathyroid. As a non-limiting example, the tumor is a benign tumor called c cell hyperplasia located in the thyroid tissue (C cells). As a non-limiting example, the tumor is a malignant tumor called medullary carcinoma of thyroid located in the thyroid tissue (C cells). As a non- limiting example, the tumor is a malignant tumor called bronchial carcinooid located in the bronchial lining (Kultschitzky cells). As a non-limiting example, the tumor is a malignant tumor called oat cells carcinoma located in the bronchial lining (Kultschitzky cells). As a non- limiting example, the tumor is a benign tumor called pheochromocytoma located in the adrenalmedulla. As a non-limiting example, the tumor is a malignant tumor called malignant pheochromocytoma located in the adrenalmedualla. As a non-limiting example, the tumor is a benign tumor called islet cell adenoma located in the pancreas. As a non-limiting example, the tumor is a benign tumor called insulinoma located in the pancreas. As a non-limiting example, the tumor is a benign tumor called gastrinoma located in the pancreas. As a non-limiting example, the tumor is a malignant tumor called islet cell carcinoma located in the pancreas. As a non-limiting example, the tumor is a benign tumor called carcinoid located in the stomach and intestines. As a non-limiting example, the tumor is a malignant tumor called malignant carcinoid located in the stomach and intestines. As a non-limiting example, the tumor is a benign tumor called chemodectoma located in the carotid body and chemo-receptor system. As a non-limiting example, the tumor is a benign tumor called paraganglioma located in the carotid body and chemo-receptor system. As a non-limiting example, the tumor a malignant tumor called malignant carcinoid located in the carotid body and chemo-receptor system. As a non- limiting example, the tumor a malignant tumor called malignant paraganglioma located in the carotid body and chemo-receptor system. [0834] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in neural crest-derived cells such as, but not limited to, pigment-producing cells (e.g., skin and eyes), schwann cells of the peripheral nervous system, and merkel cells in the squamous epithelium. As a non-limiting example, the tumor is a benign tumor called nevus located in pigment-producing cells such as the skin and eyes. As a non- limiting example, the tumor a malignant tumor called melanoma located in pigment-producing cells such as the skin and eyes. As a non-limiting example, the tumor is a benign tumor called schwannoma or neurilemmoma located in schwann cells of the peripheral nervous system. As a non-limiting example, the tumor is a malignant tumor called malignant schwannoma located in schwann cells of the peripheral nervous system. As a non-limiting example, the tumor is a malignant tumor called merkel cell neoplasm located in merkel cells in the squamous epithelium. [0835] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in breast tissue. As a non-limiting example, the tumor is a benign tumor called fibroadenoma. As a non-limiting example, the tumor is a malignant tumor called cystosarcoma phylloides. [0836] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in renal anlage tissue. As a non-limiting example, the tumor is a malignant tumor called Wilms tumor. [0837] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in ovary tissue. [0838] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in testis tissue. [0839] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in germ cell tumor tissue. Non-limiting examples of germ cell tumors including seminoma, dysgerminoma, choriocarcinoma, embryonal carcinoma, endodermal sinus tumor, and teratocarcinoma. [0840] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof is located in the connective tissue stroma. Non-limiting examples of these tumors are Sertoli-Leydig cell tumors, arrhenoblastoma, granulose-theca cell tumors, hilar cell tumors, lipid cell tumors. Organs [0841] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to an organ. Non-limiting example of organs include the anal canal, arteries, ascending colon, bladder, bone marrow, brain, bronchi, bronchioles, bulbourethral glands, capillaries, cecum, cerebellum, cerebral hemispheres, cerebrum, cervix, choroid plexus, clitoris, cranial nerves, descending colon, diencephalon, duodenum, ear, enteric nervous system, epididymis, esophagus, external reproductive organs, fallopian tubes, gallbladder, ganglia, gustatory, gut-associated lymphoid tissue, heart, ileum, internal reproductive organs, interstitium, jejunum, joints, kidneys, large intestine, larynx, ligaments, liver, lungs, lymph node, lymphatic vessel, mammary glands, medulla oblongata, mesentery, midbrain, mouth, muscles of breathing, nasal cavity, nerves, olfactory, ovaries, pancreas, parotid glands, penis, pharynx, placenta, pons, prostate, rectum, salivary glands, scrotum, seminal vesicles, sigmoid colon, skeleton, skin, small intestine, spinal nerves, spleen, stomach, subcutaneous tissue, sublingual glands, submandibular glands, teeth, tendons, testes, the brainstem, the spinal cord, the ventricular system, thymus, tongue, tonsils, trachea, transverse colon, ureter, urethra, uterus, vagina, vas deferens, veins, and vulva. Tissues [0842] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to a tissue. Non-limiting example of adrenal medulla, adult fibrous tissue, blood vessels, bone, breast, bronchial lining, carotid body, cartilage, connective tissue, embryonic (myxomatous) fibrous tissue, epithelial, epithelium, fat, glandular epithelium (liver, kidney, bile duct), gonads, hematopoietic cells, lymph vessels, lymphoid tissue, meninges, mesothelium, muscle, nerve sheath, nervous, notochord, ovary, pancreas, parathyroid, pituitary, placenta, renal anlage, smooth muscle, stomach and intestines, stratified squamous, striated muscle, stroma, testis, thyroid, and transitional epithelium. As a non- limiting example, the tissue is connective tissue. Cells [0843] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to a particular cell type. Non-limiting example of cells include adipocytes, adrenergic neural cells, alpha cell, amacrine cells, ameloblast, anterior lens epithelial cell, anterior/intermediate pituitary cells, apocrine sweat gland cell, astrocytes, auditory inner hair cells of organ of corti, auditory outer hair cells of organ of corti, b cell, bartholin's gland cell, basal cell (stem cell) of cornea, tongue, mouth, nasal cavity, distal anal canal, distal urethra, and distal vagina, basal cells of olfactory epithelium, basket cells, basophil granulocyte and precursors, beta cell, betz cells, bone marrow reticular tissue fibroblasts, border cells of organ of corti, boundary cells, bowman's gland cell, brown fat cell, brunner's gland cell, bulbourethral gland cell, bushy cells, c cells, cajal–retzius cells, cardiac muscle cell, cardiac muscle cells, cartwheel cells, cells of the zona fasciculata produce glucocorticoids, cells of the zona glomerulosa produce mineralocorticoids, cells of the zona reticularis produce androgens, cells of the adrenal cortex, cementoblast, centroacinar cell, ceruminous gland cell in ear, chandelier cells, chemoreceptor glomus cells of carotid body cell, chief cell, cholinergic neurons, chromaffin cells, club cell, cold-sensitive primary sensory neurons, connective tissue macrophage (all types), corneal fibroblasts (corneal keratocytes), corpus luteum cell of ruptured ovarian follicle secreting progesterone, cortical hair shaft cell, corticotropes, crystallin-containing lens fiber cell, cuticular hair shaft cell, cytotoxic t cell, d cell, delta cell, dendritic cell, double-bouquet cells, duct cell, eccrine sweat gland clear cell, eccrine sweat gland dark cell, efferent ducts cell, elastic cartilage chondrocyte, endothelial cells, enteric glial cells, enterochromaffin cell, enterochromaffin-like cell, enteroendocrine cell, eosinophil granulocyte and precursors, ependymal cells, epidermal basal cell, epidermal langerhans cell, epididymal basal cell, epididymal principal cell, epithelial reticular cell, epsilon cell, erythrocyte, fibrocartilage chondrocyte, fork neurons, foveolar cell, g cell, gall bladder epithelial cell, germ cells, gland of littre cell, gland of moll cell in eyelid, glial cells, golgi cells, gonadal stromal cells, gonadotropes, granule cells, granulosa cell, granulosa lutein cells, grid cells, head direction cells, and hematopoietic stem cells. In some embodiments, the at least one cell type comprise cancerous cells. In some embodiments, the at least one cell type comprise non-cancerous cells. In some embodiments, the at least one cell type comprise both cancerous and non-cancerous type. In some embodiments, the cancerous state of the at least one cell type is unknown. Physiological Systems [0844] In some embodiments, the delivery of nucleic acid sequences, polypeptides or peptides and formulations thereof can be localized to a physiological system. Non-limiting example of physiological system include the auditory, cardiovascular, central nervous system, chemo- receptor system, circulatory, digestive, endocrine, excretory, exocrine, genital, integumentary, lymphatic, muscular, musculoskeletal , nervous, peripheral nervous system, renal, reproductive, respiratory, urinary, and visual systems. VII. METHODS OF DETECTION AND ANALYSIS [0845] Detection of the tropism discovery platform including the targeting systems (e.g., candidate targeting system and validated targeting system) may be carried out through a variety of techniques (i.e., detection techniques or analysis techniques, both of which are used interchangeably herein) which can be selected based on the tracking system used. [0846] In some embodiments, the targeting systems described herein is detected utilizing a nuclear imaging technique. Nuclear imaging techniques, as used herein, are meant to encompass any imaging, detection, couniting, or sorting technique that utilizes radioactive emissions, ether emitted from the subject or an external source. Without limitation, nuclear imaging techniques may include X-ray, magnetic resonance imaging (MRI) including functional magnetic resonance imaging (fMRI) and nuclear magnetic resonance imaging, computed tomography (CT), positron emission tomography (PET), single-photon emission computed tomography (SPECT), absorption imaging, or any combination thereof. The general principles and procedures of these approaches are known in the art, see Pérez-Medina, et. al., Nuclear imaging approaches facilitating nanomedicine translation. Advanced Drug Delivery Reviews 154–155 (2020) 123–141, the contents of which are herein incorporated by reference in their entirety as it relates to nuclear imaging techniques. [0847] In some embodiments, detection of the targeting systems described herein in a subject may be performed utilizing MRI techniques. This approach may be carried out by any method known or discovered. While not wishing to be bound by theory, MRI utilizes the detection of certain nuclide spin characteristics. In some embodiments, MRI may be used as a non-invasive detection technique along with the targeting systems described herein that comprises an MRI contrast agent such as gadolinium-based small molecules, manganese-based small molecules, iron oxide nanoparticles, 19 F-based compounds, and any combination thereof. MRI techniques may, as an example, allow for a detection of the targeting systems in specific organs and tissues of a subject in vivo, as well as changes in those distributions over time. [0848] In some embodiments, detection of the targeting systems described herein in a subject may be performed utilizing CT techniques. This approach may be carried out by any method known or discovered. While not wishing to be bound by theory, CT utilizes the interaction of X-ray photons with matter, CT may be used as a non-invasive detection technique along with the targeting systems that comprise an CT contrast agent such as a gold high-density lipoprotein nanoparticle (Au-HDL). CT techniques may, as an example, allow for a detection of the targeting systems in specific organs and tissues of a subject in vivo, as well as changes in those distributions over time. [0849] In some embodiments, detection of the targeting systems described herein in a subject may be performed utilizing PET techniques. This approach may be carried out by any method known or discovered. While not wishing to be bound by theory, PET utilize detection of photon emission from exogenously administered radiological substances, i.e., radiotracers. Principally, PET scanners detect the two photons emitted in opposite directions after positron-electron annihilation (the coincidence event). PET may be used as either an invasive or non-invasive detection technique along with the targeting systems that comprise an appropriate radiolabel such as 111In, 99mTc, 13N, 68Ga, 18F, 64Cu, 86Y, 76Br, 89Zr, 72As, 124I, 74As, fluorine- 18, gallium-68, nitrogen-13, copper-64, bromine-76, iodine-125, arsenic-74, carbon-11, iodine-131, 153Sm, 177Lu, 186Re, 188Re, 198Au, and 225Ac. These labels may be conjugated to either the structural elements, the cargo components, or both. PET scans may be performed to detect distribution of the targeting systems either on the subject in vivo, including changes in those distributions over time, or on excised samples of the subject. PET techniques may allow for detection of the targeting systems in a subject from the organ/tissue level down to the cell type level. Some PET techniques may allow for detection of the targeting systems at the intracellular level. [0850] In some embodiments, detection of the targeting systems described herein in a subject may be performed utilizing SPECT techniques. This approach may be carried out by any method known or discovered. While not wishing to be bound by theory, SPECT utilize detection of photon emission from exogenously administered radiological substances, i.e., radiotracers. Principally, SPECT scanners detect X-ray and gamma photons associated with nuclear state transitions. SPECT may be used as either an invasive or non-invasive detection technique along with the targeting systems that comprise an appropriate radiolabel such as 111In, 99mTc, 13N, 68Ga, 18F, 64Cu, 86Y, 76Br, 89Zr, 72As, 124I, 74As, fluorine-18, gallium-68, nitrogen-13, copper-64, bromine-76, iodine-125, arsenic-74, carbon-11, iodine- 131, 153Sm, 177Lu, 186Re, 188Re, 198Au, and 225Ac. These labels may be conjugated to either the structural elements, the cargo components, or both. SPECT scans may be performed to detect distribution of the targeting systems either on the subject in vivo, including changes in those distributions over time, or on excised samples of the subject. SPECT techniques may allow for detection of the targeting systems in a subject from the organ/tissue level down to the cell type level. Some SPECT techniques may allow for detection of the targeting systems at the intracellular level. [0851] In some embodiments, multiple nuclear imaging techniques may be used with the targeting systems comprising a single tracking system. In some embodiments, multiple nuclear imaging techniques may be used with the targeting systems comprising multiple tracking systems. [0852] In some embodiments, the targeting systems described herein is detected utilizing an optical imaging technique. Optical imaging techniques, as used herein, are meant to encompass any imaging, detection, couniting, or sorting technique that utilizes light emissions and the special properties of photons, ether emitted from the subject or an external source. Without limitation, optical imaging techniques may include visible light microscopy, Raman spectroscopy, fluorescence microscopy, bioluminescence imaging (BLI), optical coherence tomography, or any combination thereof. The general principles and procedures of these approaches are known in the art, see Drummen. Fluorescent Probes and Fluorescence (Microscopy) Techniques — Illuminating Biological and Biomedical Research. Molecules 2012, 17, 14067-14090, Boutorine, et. al., Fluorescent Probes for Nucleic Acid Visualization in Fixed and Live Cells. Molecules 2013, 18, 15357-15397, and Juskowiak, Nucleic acid-based fluorescent probes and their analytical potential. Anal. Bioanal. Chem. (2011) 399:3157– 3176, the contents of which are herein incorporated by reference in their entirety as relates to optical imaging techniques. [0853] In some embodiments, detection of the targeting systems described herein in a subject may be performed utilizing visible fluorescence microscopy techniques. Fluorescence microscopy techniques include a wide range of techniques known in the art including without limitation confocal fluorescence microscopy, fluorescence reflectance imaging, fluorescence molecular tomographic imaging, and Förster Resonance Energy Transfer (FRET). In general, all fluorescence microscopy techniques utilize detection of light emitted from endogenously present or exogenously administered fluorescent compounds, i.e., compounds which absorb light or other electromagnetic radiation and re-emits it at longer wavelengths. Fluorescence microscopy techniques may be used as either an invasive or non-invasive detection technique along with the targeting systems that comprise at least one tracking system which comprises an appropriate fluorescent compound. Without limitation, such fluorescent compounds may include Green Florescent Protein, Yellow Florescent Protein, Red Florescent Protein, Sirius, EBFP2, CFP, Cerulean, EGFP, EYFP, mOrange, mCherry, mPlum, NIR, iRFP, EosFP, PamCherry, Dronpa, Dreiklang, asFP595, mMaple, mGeo, mEos2, Dendra2, psCFP2, 2,3,5,6- tetracarbazole-4-cyano-pyridine (CPy), florescent nanoparticles, or florescent lipids, fluorescein, TAMRA, Cy dyes, Texas red, HEX, JOE, Oregon green, rhodamine 6 G, coumarin, pyrene, DiOC6 (3,3′-dihexyloxacarbocyanine iodide), or any combination thereof. In some embodiments, a targeting system for detection with fluorescence microscopy will comprise at least one fluorophore which may include, without limitation, a quantum dot, a Coumarins, a Naphthalimide, a Fluorescein, a BODIPY, a Cyanine, a xanthene, an oxazine, an Oligothiophenes, and a Phthalocyanine derivative (PcDer). These fluorescence compounds may be incorporated into the structure of the targeting systems, loaded as a cargo or payload, expressed as the product of a cargo or payload, or any combination thereof. Fluorescence microscopy techniques may be performed to detect distribution of the targeting systems either on the subject in vivo, including changes in those distributions over time, or on excised samples of the subject. Fluorescence microscopy techniques may allow for detection of the targeting systems in a subject from the organ/tissue level down to the cell type level. Some fluorescence microscopy techniques may allow for detection of the targeting systems at the intracellular level. In some embodiments, fluorescence microscopy techniques may be used to sort samples of cells post administration utilizing Fluorescence-activated Cell Sorting (FACS). [0854] In some embodiments, detection of the targeting systems in a subject may be performed utilizing bioluminescence imaging (BLI) techniques. This approach may be carried out by any method known or discovered. While not wishing to be bound by theory, BLI imaging utilizes exogenously supplied compounds which emit light as a product of a chemical reaction under physiological condition. These emissions may be detected through various techniques of light and fluorescence microscopy. In some embodiments, BLI techniques may be used in conjunction with targeting systems which comprise bioluminescent compounds. Such compounds may be incorporated into nanoparticles or as the cargo or payload for expression post-delivery. In some embodiments, bioluminescent compounds may include, but are not limited to, luciferases including Renilla luciferase, Gaussia luciferase, Nanoluc luciferase, Firefly luciferase, Click Beetle luciferases, or any combination thereof. BLI techniques may be performed to detect distribution of the tropism discovery platform either on the subject in vivo, including changes in those distributions over time, or on excised samples of the subject. BLI may allow for detection of the targeting systems in a subject from the organ/tissue level down to the cell type level. Some BLI techniques may allow for detection of the targeting systems at the intracellular level. In some embodiments, BLI techniques may include quantifying luciferase expression from different organs with an in vivo imaging system (IVIS). [0855] In some embodiments, detection of the targeting systems described herein may be performed utilizing nucleotide sequencing techniques. Nucleotide sequencing techniques maybe used to detect the presence of a known sequence of nucleotides, such as an identifier (e.g., barcode) sequence, in a sample. Non-limiting examples of nucleotide sequencing techniques which may be used to detect the targeting systems include high throughput sequencing, PCR, deep sequencing, and any combination thereof. [0856] In some embodiments, detection of the targeting systems described herein may be performed by detecting the product of a tracking system which comprises a functional polynucleotide (e.g., DNA, mRNA, or oRNA) coding for a known peptide sequence or protein (i.e., a reporter sequence). In some embodiments, the functional polynucleotide may comprise a sequence which codes for a unique non-functional polypeptide sequence (i.e., a peptide or protein). In some embodiments, the reporter sequence may comprise a β-galactosidase (β-gal) sequence. In some embodiments, the reporter sequence may comprise a eGFP, luciferase, gene editor (e.g. cas9 edit, DNA readout), ox-40, beta6 integrin, CD45, a surface marker with (3x)- HA tag, (3x)-flag tag (with or without) a TEV protease site, or any combination thereof. In some embodiments, the reporter sequence may comprise a luciferase or fluorescent compound sequence. In some embodiments, the expression of the functional sequence, and by extension the presence of the targeting systems may be performed by any technique disclosed previously. In some embodiments, detecting the product of a tracking system which comprises a reporter sequence may be performed using any method known or discovered to detect products of expression. Such techniques include, but are not limited to, liquid/gas chromatography, mass spectrometry, light spectrometry (absorbance), gel electrophoresis, quantitative enzyme-linked immunosorbent assays (ELISA), Western blotting, dot blotting, Northern Blotting, protein immunostaining, protein immunoprecipitation, or any combination thereof. [0857] In some embodiments, detection of the targeting systems described herein may be performed by utilizing detections systems chosen to match especially designed tracking systems. As a non-limiting example, the targeting systems described herein may be detected by electron microscopy, thermal imaging, ultrasound imaging, photoacoustic imaging, lab assays, and any combination thereof. [0858] In some embodiments, detection of the targeting systems described herein may be performed by utilizing cell sorting techniques, including but not limited to, magnetic beads, flow cytometry, cleavage of peptide with LC-MS/MS, Fluorescence-activated Cell Sorting (FACS), or any combination thereof, combined with tracking system nanoparticles comprising components recognized by the cell sorting method. [0859] In some embodiments, a detection technique may analyze only one formulation or cargo at a time. In some embodiments, a detection technique may analyze multiple formulations or cargos at a time. In some embodiments, a detection technique may analyze about 1 formulation, 2 formulations, 3 formulations, 4 formulations 5, formulations, 6 formulations, 7, formulations, 8, formulations, 9 formulations, 10 formulations, 11 formulations, 12 formulations, 13 formulations, 14 formulations, 15 formulations, 16 formulations, 17 formulations, 18 formulations, 19 formulations, 20 formulations, 21 formulations, 22 formulations, 23 formulations, 24 formulations, 25 formulations, or more at a time. In some embodiments, a detection technique may analyze between about 1 and 100 formulations. As a non-limiting example, a detection technique may analyze about 1-10, 1-20, 1-30, 1-40.1-50, 1-60, 1-70.1- 80, or 1-90 formulations. In some embodiments, a detection technique may analyze more than 100 formulations at a time. [0860] In some embodiments, a library of targeting systems may be analyzed. As a non- limiting examples, targeting systems may have the same formulation and different identifier sequences or moieties. As another non-limiting example, targeting systems may have the same formulation and the same identifier sequences or moieties. As another non-limiting example, targeting systems may have different formulations and the same identifier sequence or moieties. As another non-limiting example, targeting systems may have different formulations and different identifier sequences of moieties. [0861] In some embodiments, a library of targeting systems may have one identifier sequence or moiety for analysis. [0862] In some embodiments, a library of targeting systems may have at least two identifier sequences or moieties for analysis. The library may have 2-10 identifier sequences or moieties for analysis. The library may have 2-100 identifier sequences or moieties for analysis. The library may have 2-500 identifier sequences or moieties for analysis. The library may have 100-500 identifier sequences or moieties for analysis. The library may have 2-1000 identifier sequences or moieties for analysis. The library may have 2-2500 identifier sequences or moieties for analysis. The library may have 1000-2500 identifier sequences or moieties for analysis. The library may have 1000-5000 identifier sequences or moieties for analysis. The library may have 2500-5000 identifier sequences or moieties for analysis. The library may have 4000-5000 identifier sequences or moieties for analysis. [0863] In some embodiments, a library of targeting systems may have at least one originator constructs or benchmark constructs formulated in a nanoparticle delivery vehicle. The library may have 1-10000 nanoparticles. The library may have 1-10 nanoparticles. The library may have 1-100 nanoparticles. The library may have 1-500 nanoparticles. The library may have 100-500 nanoparticles. The library may have 1-1000 nanoparticles. The library may have 100- 500 nanoparticles. The library may have 1000-5000 nanoparticles. The library may have 2500- 5000 nanoparticles. The library may have 1-5000 nanoparticles. The library may have 1-10000 nanoparticles. The library may have 5000-10000 nanoparticles. As a non-limiting example, the nanoparticle may be a lipid nanoparticle. VIII. METHODS OF USE [0864] In some embodiments, the tropism delivery systems described herein may be used as a therapeutic to diagnose, prevent, treat and/or manage disease, disorders and conditions, or as a diagnostic. The therapeutic may be used in personalized medicine, immuno-oncology, cancer, vaccines, gene editing (e.g., CRISPR). [0865] In some embodiments, the tropism delivery systems described herein may be used for diagnostic purposes or as diagnostic tools. [0866] In some emodibments, delivery systems described herein may be used to treat a foodborne illness, gastroentities, an infectious disease, a neglected topical disease, a tropical disease, a vector-borne disease, a toxin exposure, [0867] The pharmaceutical composition may be delivered as described in PCT Publication WO2012135805, which is incorporated herein by reference in its entirety. [0868] The present disclosure provides methods comprising administering a pharmaceutical composition to a subject in need thereof. The pharmaceutical composition may be administered to a subject using any amount and any route of administration which may be effective for preventing, treating, diagnosing, or imaging a disease, disorder, and/or condition. The exact amount required will vary from subject to subject, depending on factors such as, but not limited to, the species, age, and general condition of the subject, the severity of the disease, the particular composition, its mode of administration, its mode of activity, and the like. The pharmaceutical composition may be administered to animals, such as mammals (e.g., humans, domesticated animals, cats, dogs, monkeys, mice, rats, etc.). The payload of the pharmaceutical composition is a polynucelotide. [0869] In some embodiments, pharmaceutical, prophylactic, diagnostic, or imaging compositions thereof are administered to humans. [0870] In some embodiments, the active agent is administered by one or more of a variety of routes, including, but not limited to, local, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (e.g., by powders, ointments, creams, gels, lotions, and/or drops), mucosal, nasal, buccal, enteral, vitreal, intratumoral, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; as an oral spray, nasal spray, and/or aerosol, and/or through a portal vein catheter. [0871] In some embodiments, the active agent is administered by systemic intravenous injection. [0872] In some embodiments, the active agent is administered intravenously and/or orally. [0873] In specific embodiments, the active agent may be administered in a way which allows the active agent to cross the blood-brain barrier, vascular barrier, or other epithelial barrier. [0874] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing agents, wetting agents, and/or suspending agents. Sterile injectable preparations may be sterile injectable solutions, suspensions, and/or emulsions in nontoxic parenterally acceptable diluents and/or solvents, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution, U.S.P., and isotonic sodium chloride solution. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. Fatty acids such as oleic acid can be used in the preparation of injectables. [0875] Injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, and/or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. [0876] Dosage forms for local, topical and/or transdermal administration of a pharmaceutical composition may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and/or patches. Additionally, the present disclosure contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms may be prepared, for example, by dissolving and/or dispensing the compound in the proper medium. Alternatively or additionally, rate may be controlled by either providing a rate controlling membrane and/or by dispersing the compound in a polymer matrix and/or gel. [0877] Formulations suitable for topical administration include, but are not limited to, liquid and/or semi liquid preparations such as liniments, lotions, oil in water and/or water in oil emulsions such as creams, ointments and/or pastes, and/or solutions and/or suspensions. Topically-administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein. [0878] A pharmaceutical composition may be prepared, packaged, and/or sold in a formulation suitable for ophthalmic administration. Such formulations may, for example, be in the form of eye drops including, for example, a 0.1/1.0% (w/w) solution and/or suspension of the active ingredient in an aqueous or oily liquid excipient. Such drops may further comprise buffering agents, salts, and/or one or more other of any additional ingredients described herein. Other opthalmically-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form and/or in a liposomal preparation. Ear drops and/or eye drops are contemplated as being within the scope of this disclosure. [0879] In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the active agent to be delivered (e.g., its stability in the environment of the gastrointestinal tract, bloodstream, etc), the condition of the patient (e.g., whether the patient is able to tolerate particular routes of administration), etc. The present disclosure encompasses the delivery of the active agent by any appropriate route taking into consideration likely advances in the sciences of drug delivery. [0880] In certain embodiments, pharmaceutical compositions in accordance with the present disclosure may be administered at dosage levels sufficient to deliver from about 0.0001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, from about 0.1 mg/kg to about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, or from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic, diagnostic or prophylactic effect. The desired dosage may be delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dosage may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations). When multiple administration is employed, split dosing regimens such as those described herein may be used. [0881] According to the present disclosure, administration of active agent in split-dose regimens may produce higher levels of proteins in mammalian subjects. As used herein, a “split dose” is the division of single unit dose or total daily dose into two or more doses. As used herein, a “single unit dose” is a dose of any therapeutic administered in one dose/at one time/single route/single point of contact, i.e., single administration event. As used herein, a “total daily dose” is an amount given or prescribed in 24 hr period. It may be administered as a single unit dose. In one embodiment, the active agent of the present disclosure are administered to a subject in split doses. In some embodiments, the active agent is formulated in buffer only or in a formulation described herein. [0882] LNPs of the present disclosure may be used or administered in combination with one or more other therapeutic, prophylactic, diagnostic, or imaging agents. By “in combination with,” it is not intended to imply that the agents must be administered at the same time and/or formulated for delivery together, although these methods of delivery are within the scope of the present disclosure. Pharmaceutical compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. In general, each agent will be administered at a dose and/or on a time schedule determined for that agent. In some embodiments, the present disclosure encompasses the delivery of pharmaceutical, prophylactic, diagnostic, or imaging compositions in combination with agents that may improve their bioavailability, reduce and/or modify their metabolism, inhibit their excretion, and/or modify their distribution within the body. [0883] It will further be appreciated that therapeutically, prophylactically, diagnostically, or imaging active agents utilized in combination may be administered together in a single pharmaceutical composition or administered separately in different pharmaceutical compositions. In general, it is expected that agents utilized in combination with be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually. In one embodiment, the combinations, each or together may be administered according to the split dosing regimens described herein. [0884] The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, a pharmaceutical composition useful for treating cancer in accordance with the present disclosure may be administered concurrently with a chemotherapeutic agent), or they may achieve different effects (e.g., control of any adverse effects). [0885] Pharmaceutical compositions containing LNPs disclosed herein are formulated for administration intramuscularly, transarterially, intraocularly, vaginally, rectally, intraperitoneally, intravenously, intranasally, subcutaneously, endoscopically, transdermally, intramuscularly, intraventricularly, intradermally, intrathecally, topically (e.g. by powders, ointments, creams, gels, lotions, and/or drops), mucosally, nasal, enterally, intratumorally, by intratracheal instillation, bronchial instillation, and/or inhalation; nasal spray and/or aerosol, and/or through a portal vein catheter. [0886] The pharmaceutical compositions may also be formulated for direct delivery to an organ or tissue in any of several ways in the art including, but not limited to, direct soaking or bathing, via a catheter, by gels, powder, ointments, creams, gels, lotions, and/or drops, by using substrates such as fabric or biodegradable materials coated or impregnated with the pharmaceutical compositions, and the like. In some embodiments, the pharmaceutical composition is formulated for extended release. In specific embodiments, the active agent and/or pharmaceutical, prophylactic, diagnostic, or imaging compositions thereof, may be administered in a way which allows the active agent to cross the blood-brain barrier, vascular barrier, or other epithelial barrier. [0887] In some aspects of the present disclosure, the active agent of the present disclosure are spatially retained within or proximal to a target tissue. Provided are methods of providing a pharmaceutical composition to a target tissue of a mammalian subject by contacting the target tissue (which contains one or more target cells) with the pharmaceutical composition under conditions such that the pharmaceutical composition, in particular the active agent component(s) of the pharmaceutical composition, is substantially retained in the target tissue, meaning that at least 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99 or greater than 99.99% of the pharmaceutical composition is retained in the target tissue. Advantageously, retention is determined by measuring the amount of a component of the active agent present in the pharmaceutical composition that enters one or more target cells. For example, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99 or greater than 99.99% of the active agent administered to the subject are present intracellularly at a period of time following administration. [0888] Aspects of the present disclosure are directed to methods of providing a pharmaceutical composition to a target tissue or organ of a mammalian subject, by contacting the target tissue (containing one or more target cells) or organ (containing one or more target cells) with the pharmaceutical composition under conditions such that the pharmaceutical composition is substantially retained in the target tissue or organ. The pharmaceutical composition contains an effective amount of an active agent. [0889] Pharmaceutical compositions which may be administered intramuscularly and/or subcutaneously may include, but are not limited to, polymers, copolymers, and gels. The polymers, copolymers and/or gels may further be adjusted to modify release kinetics by adjusting factors such as, but not limited to, molecular weight, particle size, payload and/or ratio of the monomers. As a nonlimiting example, formulations administered intramuscularly and/or subcutaneously may include a copolymer such as poly(lactic-co-glycolic acid). [0890] Localized delivery of the pharmaceutical compositions described herein may be administered by methods such as, but not limited to, topical delivery, ocular delivery, transdermal delivery, and the like. The pharmaceutical composition may also be administered locally to a part of the body not normally available for localized delivery such as, but not limited to, when a subject’s body is open to the environment during treatment. The pharmaceutical composition may further be delivered by bathing, soaking and/or surrounding the body part with the pharmaceutical composition. [0891] However, the present disclosure encompasses the delivery of an active agent disclosed herein, and/or pharmaceutical, prophylactic, diagnostic, or imaging compositions thereof, by any appropriate route taking into consideration likely advances in the sciences of drug delivery. Methods of Producing Polypeptides in Cells [0892] The present disclosure provides methods of producing a polypeptide of interest in a mammalian cell. Methods of producing polypeptides involve contacting a cell with a formulation of the disclosure comprising an LNP including an mRNA encoding the polypeptide of interest. Upon contacting the cell with the lipid nanoparticle, the mRNA may be taken up and translated in the cell to produce the polypeptide of interest. [0893] In general, the step of contacting a mammalian cell with a LNP including an mRNA encoding a polypeptide of interest may be performed in vivo, ex vivo, in culture, or in vitro. The amount of lipid nanoparticle contacted with a cell, and/or the amount of mRNA therein, may depend on the type of cell or tissue being contacted, the means of administration, the physiochemical characteristics of the lipid nanoparticle and the mRNA (e.g., size, charge, and chemical composition) therein, and other factors. In general, an effective amount of the lipid nanoparticle will allow for efficient polypeptide production in the cell. Metrics for efficiency may include polypeptide translation (indicated by polypeptide expression), level of mRNA degradation, and immune response indicators. [0894] The step of contacting an LNP including an mRNA with a cell may involve or cause transfection. A phospholipid including in the lipid component of a LNP may facilitate transfection and/or increase transfection efficiency, for example, by interacting and/or fusing with a cellular or intracellular membrane. Transfection may allow for the translation of the mRNA within the cell. [0895] In some embodiments, the lipid nanoparticles described herein may be used therapeutically. For example, an mRNA included in an LNP may encode a therapeutic polypeptide (e.g., in a translatable region) and produce the therapeutic polypeptide upon contacting and/or entry (e.g., transfection) into a cell. In other embodiments, an mRNA included in a LNP may encode a polypeptide that may improve or increase the immunity of a subject. In some embodiments, an mRNA may encode a granulocyte-colony stimulating factor or trastuzumab. [0896] In some embodiments, an mRNA included in an LNP may encode a recombinant polypeptide that may replace one or more polypeptides that may be substantially absent in a cell contacted with the lipid nanoparticle. The one or more substantially absent polypeptides may be lacking due to a genetic mutation of the encoding gene or a regulatory pathway thereof. Alternatively, a recombinant polypeptide produced by translation of the mRNA may antagonize the activity of an endogenous protein present in, on the surface of, or secreted from the cell. An antagonistic recombinant polypeptide may be desirable to combat deleterious effects caused by activities of the endogenous protein, such as altered activities or localization caused by mutation. In another alternative, a recombinant polypeptide produced by translation of the mRNA may indirectly or directly antagonize the activity of a biological moiety present in, on the surface of, or secreted from the cell. Antagonized biological moieties may include, but are not limited to, lipids (e.g., cholesterol), lipoproteins (e.g., low density lipoprotein), nucleic acids, carbohydrates, and small molecule toxins. Recombinant polypeptides produced by translation of the mRNA may be engineered for localization within the cell, such as within a specific compartment such as the nucleus, or may be engineered for secretion from the cell or for translocation to the plasma membrane of the cell. [0897] In some embodiments, contacting a cell with an LNP including an mRNA may reduce the innate immune response of a cell to an exogenous nucleic acid. A cell may be contacted with a first lipid nanoparticle including a first amount of a first exogenous mRNA including a translatable region and the level of the innate immune response of the cell to the first exogenous mRNA may be determined. Subsequently, the cell may be contacted with a second composition including a second amount of the first exogenous mRNA, the second amount being a lesser amount of the first exogenous mRNA compared to the first amount. Alternatively, the second composition may include a first amount of a second exogenous mRNA that is different from the first exogenous mRNA. The steps of contacting the cell with the first and second compositions may be repeated one or more times. Additionally, efficiency of polypeptide production (e.g., translation) in the cell may be optionally determined, and the cell may be re-contacted with the first and/or second composition repeatedly until a target protein production efficiency is achieved. Methods of Delivering Therapeutic Agents to Cells and Organs [0898] Provided herein are methods of treating a disease or disorder, the methods comprising administering to a subject in need thereof a pharmaceutical composition of the present disclosure, such as a pharmaceutical composition comprising an LNP described herein. [0899] The present disclosure provides methods of delivering an active agent and/or prophylactic, such as a nucleic acid, to a mammalian cell or organ. Delivery of a therapeutic and/or prophylactic to a cell involves administering a formulation of the disclosure that comprises a LNP including the therapeutic and/or prophylactic, such as a nucleic acid, to a subject, where administration of the composition involves contacting the cell with the composition. In some embodiments, a protein, cytotoxic agent, radioactive ion, chemotherapeutic agent, or nucleic acid (such as an RNA, e.g., mRNA) may be delivered to a cell or organ. In the instance that a therapeutic and/or prophylactic is an mRNA, upon contacting a cell with the lipid nanoparticle, a translatable mRNA may be translated in the cell to produce a polypeptide of interest. However, mRNAs that are substantially not translatable may also be delivered to cells. Substantially non-translatable mRNAs may be useful as vaccines and/or may sequester translational components of a cell to reduce expression of other species in the cell. [0900] In some embodiments, an LNP may target a particular type or class of cells (e.g., cells of a particular organ or system thereof). In some embodiments, a LNP including a therapeutic and/or prophylactic of interest may be specifically delivered to a mammalian liver, kidney, spleen, femur, or lung. “Specific delivery” to a particular class of cells, an organ, or a system or group thereof implies that a higher proportion of lipid nanoparticles including a therapeutic and/or prophylactic are delivered to the destination (e.g., tissue) of interest relative to other destinations, e.g., upon administration of an LNP to a mammal. In some embodiments, specific delivery may result in a greater than 2 fold, 5 fold, 10 fold, 15 fold, or 20 fold increase in the amount of therapeutic and/or prophylactic per 1 g of tissue of the targeted destination (e.g., tissue of interest, such as a liver) as compared to another destination (e.g., the spleen). In some embodiments, the tissue of interest is selected from the group consisting of a liver, kidney, a lung, a spleen, a femur, vascular endothelium in vessels (e.g., intra-coronary or intra-femoral) or kidney, and tumor tissue (e.g., via intratumoral injection). [0901] As another example of targeted or specific delivery, an mRNA that encodes a protein- binding partner (e.g., an antibody or functional fragment thereof, a scaffold protein, or a peptide) or a receptor on a cell surface may be included in an LNP. An mRNA may additionally or instead be used to direct the synthesis and extracellular localization of lipids, carbohydrates, or other biological moieties. Alternatively, other therapeutics and/or prophylactics or elements (e.g., lipids or ligands) of an LNP may be selected based on their affinity for particular receptors (e.g., low density lipoprotein receptors) such that a LNP may more readily interact with a target cell population including the receptors. In some embodiments, ligands may include, but are not limited to, members of a specific binding pair, antibodies, monoclonal antibodies, Fv fragments, single chain Fv (scFv) fragments, Fab' fragments, F(ab')2 fragments, single domain antibodies, camelized antibodies and fragments thereof, humanized antibodies and fragments thereof, and multivalent versions thereof; multivalent binding reagents including mono- or bi- specific antibodies such as disulfide stabilized Fv fragments, scFv tandems, diabodies, tribodies, or tetrabodies; and aptamers, receptors, and fusion proteins. [0902] In some embodiments, a ligand may be a surface-bound antibody, which can permit tuning of cell targeting specificity. This is especially useful since highly specific antibodies can be raised against an epitope of interest for the desired targeting site. In some embodiments, multiple antibodies are expressed on the surface of a cell, and each antibody can have a different specificity for a desired target. Such approaches can increase the avidity and specificity of targeting interactions. [0903] A ligand can be selected, e.g., by a person skilled in the biological arts, based on the desired localization or function of the cell. In some embodiments an estrogen receptor ligand, such as tamoxifen, can target cells to estrogen-dependent breast cancer cells that have an increased number of estrogen receptors on the cell surface. Other non-limiting examples of ligand/receptor interactions include CCR1 (e.g., for treatment of inflamed joint tissues or brain in rheumatoid arthritis, and/or multiple sclerosis), CCR7, CCR8 (e.g., targeting to lymph node tissue), CCR6, CCR9, CCR10 (e.g., to target to intestinal tissue), CCR4, CCR10 (e.g., for targeting to skin), CXCR4 (e.g., for general enhanced transmigration), HCELL (e.g., for treatment of inflammation and inflammatory disorders, bone marrow), Alpha4beta7 (e.g., for intestinal mucosa targeting), and VLA-4NCAM-1 (e.g., targeting to endothelium). In general, any receptor involved in targeting (e.g., cancer metastasis) can be harnessed for use in the methods and compositions described herein. [0904] Targeted cells may include, but are not limited to, hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, lung cells, bone cells, stem cells, mesenchymal cells, neural cells, cardiac cells, adipocytes, vascular smooth muscle cells, cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, and tumor cells. [0905] In some embodiments, an LNP may target hepatocytes. Apolipoproteins such as apolipoprotein E (apoE) have been shown to associate with neutral or near neutral lipid- containing lipid nanoparticles in the body, and are known to associate with receptors such as low-density lipoprotein receptors (LDLRs) found on the surface of hepatocytes. Thus, an LNP including a lipid component with a neutral or near neutral charge that is administered to a subject may acquire apoE in a subject's body and may subsequently deliver a therapeutic and/or prophylactic (e.g., an RNA) to hepatocytes including LDLRs in a targeted manner. Methods of Treating Diseases and Disorders [0906] Lipid nanoparticles are useful for treating a disease, disorder, or condition. In particular, such compositions are useful in treating a disease, disorder, or condition characterized by missing or aberrant protein or polypeptide activity. In some embodiments, a formulation of the disclosure that comprises an LNP including an mRNA encoding a missing or aberrant polypeptide may be administered or delivered to a cell. Subsequent translation of the mRNA may produce the polypeptide, thereby reducing or eliminating an issue caused by the absence of or aberrant activity caused by the polypeptide. Because translation may occur rapidly, the methods and compositions may be useful in the treatment of acute diseases, disorders, or conditions such as sepsis, stroke, and myocardial infarction. A therapeutic and/or prophylactic included in an LNP may also be capable of altering the rate of transcription of a given species, thereby affecting gene expression. [0907] Diseases, disorders, and/or conditions characterized by dysfunctional or aberrant protein or polypeptide activity for which a composition may be administered include, but are not limited to, rare diseases, infectious diseases (as both vaccines and therapeutics), cancer and proliferative diseases, genetic diseases (e.g., cystic fibrosis), autoimmune diseases, diabetes, neurodegenerative diseases, cardio- and reno-vascular diseases, and metabolic diseases. Multiple diseases, disorders, and/or conditions may be characterized by missing (or substantially diminished such that proper protein function does not occur) protein activity. Such proteins may not be present, or they may be essentially non-functional. A specific example of a dysfunctional protein is the missense mutation variants of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which produce a dysfunctional protein variant of CFTR protein, which causes cystic fibrosis. The present disclosure provides a method for treating such diseases, disorders, and/or conditions in a subject by administering a LNP including an RNA and a lipid component including a PEGylated lipid compound disclosed herein, a phospholipid (optionally unsaturated), optionally a second PEGylated lipid, and a structural lipid, wherein the RNA may be an mRNA encoding a polypeptide that antagonizes or otherwise overcomes an aberrant protein activity present in the cell of the subject. [0908] In some embodiments, lipid nanoparticles disclosed herein comprise a polynucleotide encoding an antigen protein. In some embodiments, a polynucleotide is an mRNA or circRNA encoding an antigen protein. In some embodiments, a polynucleotide encodes a protein selected from SEQ ID NOs: 1-54, or a sequence having about 60% sequence identity, about 70% sequence identity, about 80% sequence identity, about 90% sequence identity, or about 95% sequence identity to a protein selected from SEQ ID NOs: 1-54. [0909] The present application contains a Sequence Listing which has been submitted electronically in XML format. The Sequence Listing XML is hereby incorporated by reference in its entirety. Said XML file, created on September 13, 2022, is named REG006_SL and is 97,563 bytes in size. [0910] In some embodiments, lipid nanoparticles disclosed herein a useful in method of treating a disease or disorder. In some embodiments, a disease or disorder is a foodbourne illness or gastroenteristis. In some embodiments, a foodbourne illness is caused by a pathogen selected from the group consisitng of Campylobacter jejuni bacteria, Clostridium difficile bacteria, Entamoeba histolytica, Enterotoxin B, Norwalk virus/Norovirus, Helicobacter pyroli, and Rotavirus. [0911] In some embodiments, a disease or disorder is an infectious disease. In some embodiments, an infectious agent is the result of an infection with an agent selected from the group consisting of candida yeast, a coronavirus (e.g., SARS-CoV, SARS-CoV-2, MERS- CoV), enterovirus 71, Epstein-Barr virus, Gram-Negative Bacteria (e.g., Bordetella), Gram- Positive bacteria (e.g., Clostridium Tetani, Francisella tularensis, Streptococcus bacteria, Staphylococcus bacteria), hepatitis virus, human cytomegalovirus, HIV, HPV, influenza virus, JCV, mycobacterium, poxviruses, pseudomonos aeruginosa, toxoplasma gondii, vaicella zoster virus, chikungunya virus, dengue virus, rabies virus, typanosoma cruzi, ebola virus, plasmodium falciparum, marbug virus, Japanese encephalitis virus, St. Louis encephalitis virus, West Nile Virus, and Yellow Fever virus. Preventative Applications [0912] In some embodiments, the tropism delivery systems described herein may be used to prevent disease or stabilize the progression of disease. [0913] In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to prevent a disease or disorder in the future. [0914] In some embodiments, the tropism delivery systems described herein may be used to halt further progression of a disease or disorder. Vaccine [0915] In some embodiments, the tropism delivery systems described herein may be used as, and/or in a manner similar to that of a vaccine. As used herein, a "vaccine" is a biological preparation that improves immunity to a particular disease or infectious agent. [0916] In some embodiments, the tropism delivery systems described herein may be used as, and/or in a manner similar to that of a vaccine for a therapeutic area such as, but not limited to, cardiovascular, CNS, dermatology, endocrinology, oncology, immunology, respiratory, and anti-infective. In some embodiments, the tropism delivery systems described herein may be used as a vaccine to diagnose, prevent, treat and/or manage a foodborne illness. In some embodiments, the tropism delivery systems described herein may be used as a vaccine to diagnose, prevent, treat and/or manage gastroenteritis. In some embodiments, the tropism delivery systems described herein may be used as a vaccine to diagnose, prevent, treat and/or manage influenza. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage HIV. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage coronavirus. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage COVID-19. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage polio. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage tetanus. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Hepatitis A. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Hepatitis B. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Hepatitis C. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Rubella. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Hib (Haemophilus influenzae type b). In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Measles. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Pertussis (Whooping Cough). In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Pneumococcal Disease. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Rotavirus. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Mumps. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Chickenpox. In some embodiments, the tropism delivery systems described herein may be used as a prophylactic to diagnose, prevent, treat and/or manage Diphtheria. e [0917] In some embodiments, the tropism delivery systems described herein may be used as, and/or in a manner similar to that of a contraceptive. As used herein, the term, "contraceptive" may be defined as any agent or method that may be used to prevent pregnancy. [0918] In some embodiments, the contraceptive may be used short-term or long-term. [0919] In some embodiments, the contraceptive may be reversible or permanent. s [0920] In some embodiments, the tropism delivery systems described herein may be used for diagnostic purposes or as diagnostic tools for any of the aforementioned diseases or disorders. [0921] In some embodiments, the tropism delivery systems described herein may be used to detect a biomarker for disease diagnosis. [0922] In some embodiments, the tropism delivery systems described herein may be used for diagnostic imaging purposes, e.g., MRI, PET, CT or ultrasound. Research [0923] In some embodiments, the tropism delivery systems described herein may be used for diagnostic purposes or as research tools for any of the aforementioned diseases or disorders. [0924] In some embodiments, the tropism delivery systems described herein may be used to detect a biomarker for research. [0925] In some embodiments, the tropism delivery systems described herein may be used in any research experiment, e.g., in vivo or in vitro experiments. [0926] In some embodiments, the tropism delivery systems described herein may be used in cultured cells. The cultured cells may be derived from any origin known to one with skill in the art, and may be as non-limiting examples, derived from a stable cell line, an animal model or a human patient or control subject. [0927] In some embodiments, the tropism delivery systems described herein may be used in in vivo experiments in animal models (i.e., mouse, rat, rabbit, dog, cat, non-human primate, guinea pig, ferret, c-elegans, drosophila, zebrafish, or any other animal used for research purposes, known in the art). [0928] In some embodiments, the tropism delivery systems described herein may be used in human research experiments or human clinical trials. [0929] In some embodiments, the tropism delivery systems described herein may be used in stem cells and/or cell differentiation IX. ENUMERATED EMBODIMENTS [0930] In an embodiment of the disclosure, provided herein is a pharmaceutical composition comprising: [0931] a) a polynucleotide encoding at least one protein of interest, and [0932] b) a delivery vehicle comprising at least one lipid [0933] wherein the composition elicits an immune response in a subject. [0934] In an embodiment, the polynucleotides are DNA. [0935] In an embodiment, the polynucleotides are RNA. [0936] In an embodiment, the RNA are short interfering RNA (siRNA). [0937] In an embodiment, the siRNA inhibits or suppresses the expression of a target of interest in a cell. [0938] In an embodiment, the inhibition or suppression is about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% and 100%, or at least 20-30%, 20-40%, 20-50%, 20-60%, 20-70%, 20- 80%, 20-90%, 20-95%, 20-100%, 30-40%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30- 95%, 30-100%, 40-50%, 40-60%, 40-70%, 40-80%, 40-90%, 40-95%, 40-100%, 50-60%, 50- 70%, 50-80%, 50-90%, 50-95%, 50-100%, 60-70%, 60-80%, 60-90%, 60-95%, 60-100%, 70- 80%, 70-90%, 70-95%, 70-100%, 80-90%, 80-95%, 80-100%, 90-95%, 90-100% or 95-100%. [0939] In an embodiment, the polynucleotides are substantially circular. [0940] In an embodiment, polynucleotide comprises an internal ribosome entry site (IRES) sequence that is operably linked to the payload sequence region. [0941] In an embodiment, the IRES sequence comprises a sequence derived from picornavirus complementary DNA, encephalomyocarditis virus (EMCV) complementary DNA, poliovirus complementary DNA, or an Antennapedia gene from Drosophila melanogaster. [0942] In an embodiment, the polynucleotide comprises a termination element, wherein the termination element comprises at least one stop codon. [0943] In an embodiment, the polynucleotide comprises a regulatory element. [0944] In an embodiment, the polynucleotide comprises at least one masking agent. [0945] In an embodiment, the substantially circular polynucleotide is produced using in vitro transcription. [0946] In an embodiment, the payload sequence region comprises a non-coding nucleic acid sequence. [0947] In an embodiment, the payload sequence region comprises a coding nucleic acid sequence. [0948] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Campylobacter jejuni. [0949] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Clostridium difficile. [0950] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Entamoeba histolytica. [0951] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for enterotoxin B. [0952] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Norwalk virus or norovirus. [0953] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Helicobacter pylori. [0954] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for rotavirus. [0955] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for candida yeast. [0956] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for coronavirus. [0957] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for SARS-CoV. [0958] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for SARS-CoV-2. [0959] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for MERS-CoV. [0960] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Enterovirus 71. [0961] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Epstein-Barr virus. [0962] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Gram-Negative Bacteria. [0963] In an embodiment, the Gram-Negative Bacteria is Bordetella. [0964] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Gram-Positive Bacteria. [0965] In an embodiment, the Gram-Positive Bacteria is Clostridium tetani. [0966] In an embodiment, the Gram-Positive Bacteria is Francisella tularensis. [0967] In an embodiment, the Gram-Positive Bacteria is Streptococcus bacteria. [0968] In an embodiment, the Gram-Positive Bacteria is Staphylococcus bacteria. [0969] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Hepatitis. [0970] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Human Cytomegalovirus. [0971] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Human Immunodeficiency Virus. [0972] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Human Papilloma Virus. [0973] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Influenza. [0974] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for John Cunningham Virus. [0975] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Mycobacterium. [0976] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Poxviruses. [0977] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Pseudomonas aeruginosa. [0978] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Respiratory Syncytial Virus. [0979] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Rubella virus. [0980] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Varicella zoster virus. [0981] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Chikungunya virus. [0982] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Dengue virus. [0983] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Rabies virus. [0984] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Trypanosoma cruzi and/or Chagas disease. [0985] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Ebola virus. [0986] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Plasmodium falciparum. [0987] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Marburg virus. [0988] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Japanese encephalitis virus. [0989] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for St. Louis encephalitis virus. [0990] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for West Nile Virus. [0991] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Yellow Fever virus. [0992] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Bacillus anthracis. [0993] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Botulinum toxin. [0994] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Ricin. [0995] In an embodiment, the coding nucleic acid sequence encodes a protein of interest for Shiga toxin and/or Shiga-like toxin. [0996] In an embodiment, the polynucleotide comprises at least one modification. [0997] In an embodiment, the at least one modification is pyridin-4-one ribonucleoside, 5-aza- uridine, 2-thio-5-aza-uridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5- hydroxyuridine, 3-methyluridine, 5-carboxymethyl-uridine, 1-carboxymethyl-pseudouridine, 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl- pseudouridine, 5-taurinomethyl-2-thio-uridine, 1-taurinomethyl-4-thio-uridine, 5-methyl- uridine, 1-methyl-pseudouridine, 4-thio-1-methyl-pseudouridine, 2-thio-1-methyl- pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2- methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, and 4-methoxy-2-thio- pseudouridine, 5-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine, N4-acetylcytidine, 5- formylcytidine, N4-methylcytidine, 5-hydroxymethylcytidine, 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio- pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza- pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5- methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2- methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, and 4-methoxy-1-methyl- pseudoisocytidine, 2-aminopurine, 2, 6-diaminopurine, 7-deaza-adenine, 7-deaza-8-aza- adenine, 7-deaza-2-aminopurine, 7-deaza-8-aza-2-aminopurine, 7-deaza-2,6-diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyladenosine, N6-methyladenosine, N6- isopentenyladenosine, N6-(cis-hydroxyisopentenyl)adenosine, 2-methylthio-N6-(cis- hydroxyisopentenyl) adenosine, N6-glycinylcarbamoyladenosine, N6- threonylcarbamoyladenosine, 2-methylthio-N6-threonyl carbamoyladenosine, N6,N6- dimethyladenosine, 7-methyladenine, 2-methylthio-adenine, and 2-methoxy-adenine, inosine, 1-methyl-inosine, wyosine, wybutosine, 7-deaza-guanosine, 7-deaza-8-aza-guanosine, 6-thio- guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine, 6- thio-7-methyl-guanosine, 7-methylinosine, 6-methoxy-guanosine, 1-methylguanosine, N2- methylguanosine, N2,N2-dimethylguanosine, 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1- methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, or N2,N2-dimethyl-6-thio-guanosine. [0998] In an embodiment, the pharmaceutical composition comprises at least one cationic lipid selected from the group consisting of any lipid in Table (I), any lipid having a structure of Formula (CY-I), any lipid having a structure of Formula (CY-II), any lipid having a structure of Formula (CY-III), any lipid having a structure of Formula (CY-IV), and combinations thereof. [0999] In an embodiment, the cationic lipid is any lipid having a structure of Formula (CY-I). [1000] In an embodiment, the cationic lipid is selected from the group consisting of Compounds CY1, CY2, CY3, CY9, CY10, CY11, CY12, CY22, CY23, CY24, CY30, CY31, CY32, CY33, CY43, CY44, CY45, CY50, CY51, CY52, and CY53. [1001] In an embodiment, the cationic lipid is any lipid having a structure of Formula (CY-II). [1002] In an embodiment, the cationic lipid is selected from the group consisting of Compounds CY4, CY5, CY16, CY17, CY18, CY25, CY26, CY37, CY38, CY39, CY46, CY56, and CY57. [1003] In an embodiment, the cationic lipid is any lipid having a structure of Formula (CY- III). [1004] In an embodiment, the cationic lipid is selected from the group consisting of Compounds CY6, CY14, CY27, CY35, CY47, and CY55. [1005] In an embodiment, the cationic lipid is any lipid having a structure of Formula (CY- IV). [1006] In an embodiment, the cationic lipid is selected from the group consisting of Compounds CY7, CY8, CY19, CY20, CY21, CY28, CY29, CY40, CY41, CY42, CY48, CY49, CY58, CY59, and CY60. [1007] In an embodiment, the pharmaceutical composition comprises an additional cationic lipid. [1008] In an embodiment, the pharmaceutical composition comprises a neutral lipid. [1009] In an embodiment, the pharmaceutical composition comprises an anionic lipid. [1010] In an embodiment, the pharmaceutical composition comprises a helper lipid. [1011] In an embodiment, the pharmaceutical composition comprises a stealth lipid. [1012] In an embodiment, the weight ratio of the lipids and the polynucleotide is between 100:1 to 1:1. [1013] In an embodiment, the pharmaceutical compositions disclosed herein preferentially target immune cells, e.g., T cells, e.g., T regulatory cells. For example, the disclosed pharmaceutical compositions may preferentially target immune cells over hepatocytes. Exemplary non-limiting immune cells include CD8+, CD4+, or CD8+CD4+ cells. In some embodiments, the disclosed pharmaceutical compositions deliver the cargo or payload to immune cells without the need for peptide-, protein-, or aptamer-based targeting ligands. See, e.g., WO 2021/021634. [1014] In an embodiment, the pharmaceutical compositions disclosed herein are delivered in the absence of a targeting ligand to mammalian liver immune cells, spleen T cells, or lung endothelial cells. Specific delivery to a particular class or type of cells indicates that a higher proportion of the pharmaceutical composition is delivered to target type or class of cells. For example, specific delivery may result in a greater than 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, or 20-fold increase in the amount of cargo or payload per 1 g of tissue of the targeted destination. [1015] In an embodiment, a vaccine formulation comprises the pharmaceutical composition. [1016] In an embodiment, a vaccine is prepared with any of Formulas (I) – (VI). [1017] In an embodiment, provided herein is a method of vaccinating a subject against an infectious agent comprising contacting a subject with the vaccine formulation or preparation and eliciting an immune response. [1018] In an embodiment, the infectious agent is Campylobacter jejuni, Clostridium difficile, Entamoeba histolytica, enterotoxin B, Norwalk virus or norovirus, Helicobacter pylori, rotavirus, candida yeast, coronavirus including SARS-CoV, SARS-CoV-2 and MERS-CoV, Enterovirus 71, Epstein-Barr virus, Gram-Negative Bacteria including Bordetella, Gram- Positive Bacteria including Clostridium tetani, Francisella tularensis, Streptococcus bacteria and Staphylococcus bacteria, and Hepatitis, Human Cytomegalovirus, Human Immunodeficiency Virus, Human Papilloma Virus, Influenza, John Cunningham Virus, Mycobacterium, Poxviruses, Pseudomonas aeruginosa, Respiratory Syncytial Virus, Rubella virus, Varicella zoster virus, Chikungunya virus, Dengue virus, Rabies virus, Trypanosoma cruzi and/or Chagas disease, Ebola virus, Plasmodium falciparum, Marburg virus, Japanese encephalitis virus, St. Louis encephalitis virus, West Nile Virus, Yellow Fever virus, Bacillus anthracis, Botulinum toxin, Ricin, or Shiga toxin and/or Shiga-like toxin.. [1019] The present disclosure includes the following enumerated embodiments 1. A pharmaceutical composition comprising: a) a polynucleotide encoding at least one protein of interest, and b) a delivery vehicle comprising at least one lipid wherein the composition elicits an immune response in a subject. 2. The pharmaceutical composition of embodiment 1, wherein the polynucleotides are DNA. 3. The pharmaceutical composition of embodiment 1, wherein the polynucleotides are RNA. 4. The pharmaceutical composition of embodiment 3, wherein the RNA are short interfering RNA (siRNA). 5. The pharmaceutical composition of embodiment 4, wherein the siRNA inhibits or suppresses the expression of a target of interest in a cell. 6. The pharmaceutical composition of embodiment 5, wherein the inhibition or suppression is about 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95% and 100%, or at least 20-30%, 20-40%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-95%, 20-100%, 30-40%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-95%, 30-100%, 40-50%, 40-60%, 40-70%, 40-80%, 40-90%, 40-95%, 40-100%, 50-60%, 50-70%, 50-80%, 50-90%, 50-95%, 50-100%, 60-70%, 60-80%, 60-90%, 60-95%, 60-100%, 70-80%, 70-90%, 70-95%, 70-100%, 80-90%, 80-95%, 80-100%, 90-95%, 90-100% or 95-100%. 7. The pharmaceutical composition of embodiments 2 or 3, wherein the polynucleotides are substantially circular. 8. The pharmaceutical composition of embodiment 7, wherein polynucleotide comprises an internal ribosome entry site (IRES) sequence that is operably linked to the payload sequence region. 9. The pharmaceutical composition of embodiment 8, wherein the IRES sequence comprises a sequence derived from picornavirus complementary DNA, encephalomyocarditis virus (EMCV) complementary DNA, poliovirus complementary DNA, or an Antennapedia gene from Drosophila melanogaster. 10. The pharmaceutical composition of embodiment 7, wherein the polynucleotide comprises a termination element, wherein the termination element comprises at least one stop codon. 11. The pharmaceutical composition of embodiment 7, wherein the polynucleotide comprises a regulatory element. 12. The pharmaceutical composition of any of embodiments 7-11, wherein the polynucleotide comprises at least one masking agent. 13. The pharmaceutical composition of any of embodiments 7-12, wherein the substantially circular polynucleotide is produced using in vitro transcription. 14. The pharmaceutical composition of any of embodiments 7-14, wherein the payload sequence region comprises a non-coding nucleic acid sequence. 15. The pharmaceutical composition of any of embodiments 7-13, wherein the payload sequence region comprises a coding nucleic acid sequence. 16. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Campylobacter jejuni. 17. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Clostridium difficile. 18. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Entamoeba histolytica. 19. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for enterotoxin B. 20. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Norwalk virus or norovirus. 21. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Helicobacter pylori. 22. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for rotavirus. 23. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for candida yeast. 24. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for coronavirus. 25. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for SARS-CoV. 26. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for SARS-CoV-2. 27. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for MERS-CoV. 28. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Enterovirus 71. 29. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Epstein-Barr virus. 30. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Gram-Negative Bacteria. 31. The pharmaceutical composition of embodiment 30, wherein the Gram-Negative Bacteria is Bordetella. 32. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Gram-Positive Bacteria. 33. The pharmaceutical composition of embodiment 32, wherein the Gram-Positive Bacteria is Clostridium tetani. 34. The pharmaceutical composition of embodiment 32, wherein the Gram-Positive Bacteria is Francisella tularensis. 35. The pharmaceutical composition of embodiment 32, wherein the Gram-Positive Bacteria is Streptococcus bacteria. 36. The pharmaceutical composition of embodiment 32, wherein the Gram-Positive Bacteria is Staphylococcus bacteria. 37. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Hepatitis. 38. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Human Cytomegalovirus. 39. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Human Immunodeficiency Virus. 40. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Human Papilloma Virus. 41. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Influenza. 42. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for John Cunningham Virus. 43. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Mycobacterium. 44. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Poxviruses. 45. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Pseudomonas aeruginosa. 46. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Respiratory Syncytial Virus. 47. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Rubella virus. 48. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Varicella zoster virus. 49. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Chikungunya virus. 50. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Dengue virus. 51. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Rabies virus. 52. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Trypanosoma cruzi and/or Chagas disease. 53. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Ebola virus. 54. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Plasmodium falciparum. 55. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Marburg virus. 56. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Japanese encephalitis virus. 57. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for St. Louis encephalitis virus. 58. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for West Nile Virus. 59. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Yellow Fever virus. 60. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Bacillus anthracis. 61. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Botulinum toxin. 62. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Ricin. 63. The pharmaceutical composition of embodiment 15, wherein the coding nucleic acid sequence encodes a protein of interest for Shiga toxin and/or Shiga-like toxin. 64. The pharmaceutical composition of any one of embodiments 3-63, wherein the polynucleotide comprises at least one modification. 65. The pharmaceutical composition of embodiment 64, wherein at least 20% of the bases are modified. 66. The pharmaceutical composition of embodiment 64, wherein at least 30% of the bases are modified. 67. The pharmaceutical composition of embodiment 64, wherein at least 40% of the bases are modified. 68. The pharmaceutical composition of embodiment 64, wherein at least 50% of the bases are modified. 69. The pharmaceutical composition of embodiment 64, wherein at least 60% of the bases are modified. 70. The pharmaceutical composition of embodiment 64, wherein at least 70% of the bases are modified. 71. The pharmaceutical composition of embodiment 64, wherein at least 80% of the bases are modified. 72. The pharmaceutical composition of embodiment 64, wherein at least 90% of the bases are modified. 73. The pharmaceutical composition of embodiment 64, wherein at least 100% of the bases are modified. 74. The pharmaceutical composition of embodiment 64, wherein a specific base comprises at least one modification. 75. The pharmaceutical composition of embodiment 74, wherein the base is adenine. 76. The pharmaceutical composition of embodiment 75, wherein at least 20% of the adenine bases are modified. 77. The pharmaceutical composition of embodiment 75, wherein at least 30% of the adenine bases are modified. 78. The pharmaceutical composition of embodiment 75, wherein at least 40% of the adenine bases are modified. 79. The pharmaceutical composition of embodiment 75, wherein at least 50% of the adenine bases are modified. 80. The pharmaceutical composition of embodiment 75, wherein at least 60% of the adenine bases are modified. 81. The pharmaceutical composition of embodiment 75, wherein at least 70% of the adenine bases are modified. 82. The pharmaceutical composition of embodiment 75, wherein at least 80% of the adenine bases are modified. 83. The pharmaceutical composition of embodiment 75, wherein at least 90% of the adenine bases are modified. 84. The pharmaceutical composition of embodiment 75, wherein at least 100% of the adenine bases are modified. 85. The pharmaceutical composition of embodiment 74, wherein the base is guanine. 86. The pharmaceutical composition of embodiment 85, wherein at least 20% of the guanine bases are modified. 87. The pharmaceutical composition of embodiment 85, wherein at least 30% of the guanine bases are modified. 88. The pharmaceutical composition of embodiment 85, wherein at least 40% of the guanine bases are modified. 89. The pharmaceutical composition of embodiment 85, wherein at least 50% of the guanine bases are modified. 90. The pharmaceutical composition of embodiment 85, wherein at least 60% of the guanine bases are modified. 91. The pharmaceutical composition of embodiment 85, wherein at least 70% of the guanine bases are modified. 92. The pharmaceutical composition of embodiment 85, wherein at least 80% of the guanine bases are modified. 93. The pharmaceutical composition of embodiment 85, wherein at least 90% of the guanine bases are modified. 94. The pharmaceutical composition of embodiment 85, wherein at least 100% of the guanine bases are modified. 95. The pharmaceutical composition of embodiment 74, wherein the base is cytosine. 96. The pharmaceutical composition of embodiment 95, wherein at least 20% of the cytosine bases are modified. 97. The pharmaceutical composition of embodiment 95, wherein at least 30% of the cytosine bases are modified. 98. The pharmaceutical composition of embodiment 95, wherein at least 40% of the cytosine bases are modified. 99. The pharmaceutical composition of embodiment 95, wherein at least 50% of the cytosine bases are modified. 100. The pharmaceutical composition of embodiment 95, wherein at least 60% of the cytosine bases are modified. 101. The pharmaceutical composition of embodiment 95, wherein at least 70% of the cytosine bases are modified. 102. The pharmaceutical composition of embodiment 95, wherein at least 80% of the cytosine bases are modified. 103. The pharmaceutical composition of embodiment 95, wherein at least 90% of the cytosine bases are modified. 104. The pharmaceutical composition of embodiment 95, wherein at least 100% of the cytosine bases are modified. 105. The pharmaceutical composition of embodiment 74, wherein the base is uracil. 106. The pharmaceutical composition of embodiment 105, wherein at least 20% of the uracil bases are modified. 107. The pharmaceutical composition of embodiment 105, wherein at least 30% of the uracil bases are modified. 108. The pharmaceutical composition of embodiment 105, wherein at least 40% of the uracil bases are modified. 109. The pharmaceutical composition of embodiment 105, wherein at least 50% of the uracil bases are modified. 110. The pharmaceutical composition of embodiment 105, wherein at least 60% of the uracil bases are modified. 111. The pharmaceutical composition of embodiment 105, wherein at least 70% of the uracil bases are modified. 112. The pharmaceutical composition of embodiment 105, wherein at least 80% of the uracil bases are modified. 113. The pharmaceutical composition of embodiment 105, wherein at least 90% of the uracil bases are modified. 114. The pharmaceutical composition of embodiment 105, wherein at least 100% of the uracil bases are modified. 115. The pharmaceutical composition of any of embodiments 64-114, wherein the at least one modification is pyridin-4-one ribonucleoside, 5-aza-uridine, 2-thio-5-aza-uridine, 2- thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine, 5-carboxymethyl-uridine, 1-carboxymethyl-pseudouridine, 5-propynyl-uridine, 1-propynyl- pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2- thio-uridine, 1-taurinomethyl-4-thio-uridine, 5-methyl-uridine, 1-methyl-pseudouridine, 4- thio-1-methyl-pseudouridine, 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza- pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2- methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, and 4-methoxy-2-thio- pseudouridine, 5-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine, N4-acetylcytidine, 5- formylcytidine, N4-methylcytidine, 5-hydroxymethylcytidine, 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio- pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza- pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5- methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2- methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, and 4-methoxy-1-methyl- pseudoisocytidine, 2-aminopurine, 2, 6-diaminopurine, 7-deaza-adenine, 7-deaza-8-aza- adenine, 7-deaza-2-aminopurine, 7-deaza-8-aza-2-aminopurine, 7-deaza-2,6-diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyladenosine, N6-methyladenosine, N6- isopentenyladenosine, N6-(cis-hydroxyisopentenyl)adenosine, 2-methylthio-N6-(cis- hydroxyisopentenyl) adenosine, N6-glycinylcarbamoyladenosine, N6- threonylcarbamoyladenosine, 2-methylthio-N6-threonyl carbamoyladenosine, N6,N6- dimethyladenosine, 7-methyladenine, 2-methylthio-adenine, and 2-methoxy-adenine, inosine, 1-methyl-inosine, wyosine, wybutosine, 7-deaza-guanosine, 7-deaza-8-aza-guanosine, 6-thio- guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine, 6-thio-7-methyl-guanosine, 7-methylinosine, 6-methoxy-guanosine, 1-methylguanosine, N2- methylguanosine, N2,N2-dimethylguanosine, 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, or N2,N2-dimethyl-6-thio- guanosine. 116. The pharmaceutical composition of embodiment 1, further comprising at least one cationic lipid selected from the group consisting of any lipid in Table (I), any lipid having a structure of Formula (CY-I), any lipid having a structure of Formula (CY-II), any lipid having a structure of Formula (CY-III), any lipid having a structure of Formula (CY-IV), and combinations thereof. 117. The pharmaceutical composition of embodiment 116, wherein the cationic lipid is any lipid having a structure of Formula (CY-I). 118. The pharmaceutical composition of embodiment 117, wherein the cationic lipid is selected from the group consisting of Compounds CY1, CY2, CY3, CY9, CY10, CY11, CY12, CY22, CY23, CY24, CY30, CY31, CY32, CY33, CY43, CY44, CY45, CY50, CY51, CY52, and CY53. 119. The pharmaceutical composition of embodiment 116, wherein the cationic lipid is any lipid having a structure of Formula (CY-II). 120. The pharmaceutical composition of embodiment 119, wherein the cationic lipid is selected from the group consisting of Compounds CY4, CY5, CY16, CY17, CY18, CY25, CY26, CY37, CY38, CY39, CY46, CY56, and CY57. 121. The pharmaceutical composition of embodiment 116, wherein the cationic lipid is any lipid having a structure of Formula (CY-III). 122. The pharmaceutical composition of embodiment 121, wherein the cationic lipid is selected from the group consisting of Compounds CY6, CY14, CY27, CY35, CY47, and CY55. 123. The pharmaceutical composition of embodiment 116, wherein the cationic lipid is any lipid having a structure of Formula (CY-IV). 124. The pharmaceutical composition of embodiment 123, wherein the cationic lipid is selected from the group consisting of Compounds CY7, CY8, CY19, CY20, CY21, CY28, CY29, CY40, CY41, CY42, CY48, CY49, CY58, CY59, and CY60. 125. The pharmaceutical composition of any of embodiments 116-124, further comprises an additional cationic lipid. 126. The pharmaceutical composition of any of embodiments 116-125, further comprising a neutral lipid. 127. The pharmaceutical composition of any of embodiments 116-126, further comprising an anionic lipid. 128. The pharmaceutical composition of any of embodiments 116-127, further comprises a helper lipid. 129. The pharmaceutical composition of any of embodiments 116-128, further comprises a stealth lipid. 130. The pharmaceutical composition of any of embodiments 116-129, wherein the weight ratio of the lipids and the polynucleotide is from about 100:1 to about 1:1. 131. A vaccine formulation comprising the pharmaceutical composition of any of embodiments 1-130. 132. A vaccine preparation comprising the pharmaceutical composition of any of embodiments 116-130. 133. A method of vaccinating a subject against an infectious agent comprising: a) contacting a subject with the vaccine formulation of embodiment 131 or the vaccine preparation of embodiment 132, and b) eliciting an immune response. 134. The method of embodiment 133, wherein the infectious agent is Campylobacter jejuni, Clostridium difficile, Entamoeba histolytica, enterotoxin B, Norwalk virus or norovirus, Helicobacter pylori, rotavirus, candida yeast, coronavirus including SARS-CoV, SARS-CoV-2 and MERS-CoV, Enterovirus 71, Epstein-Barr virus, Gram-Negative Bacteria including Bordetella, Gram-Positive Bacteria including Clostridium tetani, Francisella tularensis, Streptococcus bacteria and Staphylococcus bacteria, and Hepatitis, Human Cytomegalovirus, Human Immunodeficiency Virus, Human Papilloma Virus, Influenza, John Cunningham Virus, Mycobacterium, Poxviruses, Pseudomonas aeruginosa, Respiratory Syncytial Virus, Rubella virus, Varicella zoster virus, Chikungunya virus, Dengue virus, Rabies virus, Trypanosoma cruzi and/or Chagas disease, Ebola virus, Plasmodium falciparum, Marburg virus, Japanese encephalitis virus, St. Louis encephalitis virus, West Nile Virus, Yellow Fever virus, Bacillus anthracis, Botulinum toxin, Ricin, or Shiga toxin and/or Shiga-like toxin. 135. The method of embodiment 133, wherein the contacting is enteral (into the intestine), gastroenteral, epidural (into the dura mater), oral (by way of the mouth), transdermal, intracerebral (into the cerebrum), intracerebroventricular (into the cerebral ventricles), epicutaneous (application onto the skin), intradermal (into the skin itself), subcutaneous (under the skin), nasal administration (through the nose), intravenous (into a vein), intravenous bolus, intravenous drip, intra-arterial (into an artery), intramuscular (into a muscle), intracardiac (into the heart), intraosseous infusion (into the bone marrow), intrathecal (into the spinal canal), intraparenchymal (into brain tissue), intraperitoneal (infusion or injection into the peritoneum), intravesical infusion, intravitreal (through the eye), intracavernous injection (into a pathologic cavity) intracavitary (into the base of the penis), intravaginal administration, intrauterine, extra-amniotic administration, transdermal (diffusion through the intact skin for systemic distribution), transmucosal (diffusion through a mucous membrane), transvaginal, insufflation (snorting), sublingual, sublabial, enema, eye drops (onto the conjunctiva), ear drops, auricular (in or by way of the ear), buccal (directed toward the cheek), conjunctival, cutaneous, dental (to a tooth or teeth), electro-osmosis, endocervical, endosinusial, endotracheal, extracorporeal, hemodialysis, infiltration, interstitial, intra-abdominal, intra-amniotic, intra-articular, intrabiliary, intrabronchial, intrabursal, intracartilaginous (within a cartilage), intracaudal (within the cauda equine), intracisternal (within the cisterna magna cerebellomedularis), intracorneal (within the cornea), dental intracoronal, intracoronary (within the coronary arteries), intracorporus cavernosum (within the dilatable spaces of the corporus cavernosa of the penis), intradiscal (within a disc), intraductal (within a duct of a gland), intraduodenal (within the duodenum), intradural (within or beneath the dura), intraepidermal (to the epidermis), intraesophageal (to the esophagus), intragastric (within the stomach), intragingival (within the gingivae), intraileal (within the distal portion of the small intestine), intralesional (within or introduced directly to a localized lesion), intraluminal (within a lumen of a tube), intralymphatic (within the lymph), intramedullary (within the marrow cavity of a bone), intrameningeal (within the meninges), intramyocardial (within the myocardium), intraocular (within the eye), intraovarian (within the ovary), intrapericardial (within the pericardium), intrapleural (within the pleura), intraprostatic (within the prostate gland), intrapulmonary (within the lungs or its bronchi), intrasinal (within the nasal or periorbital sinuses), intraspinal (within the vertebral column), intrasynovial (within the synovial cavity of a joint), intratendinous (within a tendon), intratesticular (within the testicle), intrathecal (within the cerebrospinal fluid at any level of the cerebrospinal axis), intrathoracic (within the thorax), intratubular (within the tubules of an organ), intratumor (within a tumor), intratympanic (within the aurus media), intravascular (within a vessel or vessels), intraventricular (within a ventricle), iontophoresis (by means of electric current where ions of soluble salts migrate into the tissues of the body), irrigation (to bathe or flush open wounds or body cavities), laryngeal (directly upon the larynx), nasogastric (through the nose and into the stomach), occlusive dressing technique (topical route administration which is then covered by a dressing which occludes the area), ophthalmic (to the external eye), oropharyngeal (directly to the mouth and pharynx), parenteral, percutaneous, periarticular, peridural, perineural, periodontal, rectal, respiratory (within the respiratory tract by inhaling orally or nasally for local or systemic effect), retrobulbar (behind the pons or behind the eyeball), soft tissue, subarachnoid, subconjunctival, submucosal, topical, transplacental (through or across the placenta), transtracheal (through the wall of the trachea), transtympanic (across or through the tympanic cavity), ureteral (to the ureter), urethral (to the urethra), vaginal, caudal block, diagnostic, nerve block, biliary perfusion, cardiac perfusion, photopheresis, or spinal. 136. A method of delivering a polynucleotide encoding at least one protein of interest to an immune cell of a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 1-130. 137. The method of embodiment 136, wherein the immune cell is a T cell. 138. The method of embodiment 137, wherein the T cell is a CD8+ T cell. 139. The method of embodiment 137, wherein the T cell is a T regulatory cell. 140. The method of embodiment 137, wherein the T cell is CD4+ T cell. 141. The method of embodiment 136, wherein the immune cell is a macrophage, dendritic cell, or liver immune cell. X. DEFINITIONS [1020] The term "compound" or "structure," as used herein, is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. [1021] The compounds or structures described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms. [1022] Compounds or structures of the present disclosure also include tautomeric forms. Tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include prototropic tautomers which are isomeric protonation states having the same empirical formula and total charge. Examples prototropic tautomers include ketone – enol pairs, amide – imidic acid pairs, lactam – lactim pairs, amide – imidic acid pairs, enamine – imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution. [1023] Compounds or structures of the present disclosure also include all of the isotopes of the atoms occurring in the intermediate or final compounds. "Isotopes" refers to atoms having the same atomic number but different mass numbers resulting from a different number of neutrons in the nuclei. For example, isotopes of hydrogen include tritium and deuterium. [1024] The compounds or structures and salts of the present disclosure can be prepared in combination with solvent or water molecules to form solvates and hydrates by routine methods. [1025] The term "alkyl" refers to the radical of saturated aliphatic groups, including straight- chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl- substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. [1026] In some embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C30 for straight chains, C3-C30 for branched chains), 20 or fewer, 12 or fewer, or 7 or fewer. Likewise, in some embodiments cycloalkyls have from 3-10 carbon atoms in their ring structure, e.g., have 5, 6 or 7 carbons in the ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples, and claims is intended to include both "unsubstituted alkyls" and "substituted alkyls", the latter of which refers to alkyl moieties having one or more substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents include, but are not limited to, halogen, hydroxyl, carbonyl (such as a carboxyl, alkoxycarbonyl, formyl, or an acyl), thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, a hosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. [1027] Unless the number of carbons is otherwise specified, "lower alkyl" as used herein means an alkyl group, as defined above, but having from one to ten carbons, or from one to six carbon atoms in its backbone structure. Likewise, "lower alkenyl" and "lower alkynyl" have similar chain lengths. In some embodiments, alkyl groups are lower alkyls. In some embodiments, a substituent designated herein as alkyl is a lower alkyl. [1028] The term "alkylenyl" as used herein refers to a divalent radical of a straight-chain or branched-chain alkyl group. In one embodiment, the alkylenyl is a divalent form of a C 1-12 alkyl, i.e., a C1-C12 alkylenyl. In one embodiment, the alkylenyl is a divalent form of a C2-6 alkyl, i.e., a C 1 -C 10 alkylenyl. In one embodiment, the alkylenyl is a divalent form of a C 2-14 alkyl, i.e., a C1-C8 alkylenyl. In one embodiment, the alkylenyl is a divalent form of an unsubstituted C 1-6 alkyl, i.e., a C 1 -C 6 alkylenyl. In another embodiment, the alkylenyl is a divalent form of an unsubstituted C1-4 alkyl, i.e., a C1-C4 alkylenyl. Non-limiting exemplary alkylenyl groups include -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 -, - CH2(CH2)2CH2-, -CH(CH2)3CH2-, and -CH2(CH2)4CH2-. [1029] The term "cycloalkylenyl" as used herein refers to a divalent radical of a cycloalkyl group. In one embodiment, the cycloalkylenyl is a divalent form of a C 3-8 cycloalkyl, i.e., a C3-C8 cycloalkylenyl. Non-limiting exemplary cycloalkylenyl groups include: . [1030] It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For instance, the substituents of a substituted alkyl may include halogen, hydroxy, nitro, thiols, amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxylates, and esters), -CF 3 , -CN and the like. Cycloalkyls can be substituted in the same manner. [1031] The term "heteroalkyl", as used herein, refers to straight or branched chain, or cyclic carbon-containing radicals, or combinations thereof, containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B, and S, wherein the phosphorous and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroalkyls can be substituted as defined above for alkyl groups. [1032] The term "alkylthio" refers to an alkyl group, as defined above, having a sulfur radical attached thereto. In some embodiments, the "alkylthio" moiety is represented by one of -S- alkyl, -S-alkenyl, and -S-alkynyl. Representative alkylthio groups include methylthio, and ethylthio. The term "alkylthio" also encompasses cycloalkyl groups, alkene and cycloalkene groups, and alkyne groups. "Arylthio" refers to aryl or heteroaryl groups. Alkylthio groups can be substituted as defined above for alkyl groups. [1033] The terms "alkenyl" and "alkynyl", refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively. In one embodiment, the alkenyl contains one double bond. In another embodiment, the alkenyl contains two double bonds. In another embodiment, the alkenyl contains three double bonds. [1034] The term "alkenylenyl" as used herein refers to a divalent radical of an alkenyl group. In one embodiment, the alkenylenyl is a divalent form of a C 2-12 alkenyl, i.e., a C 2 -C 12 alkenylenyl. In one embodiment, the alkenylenyl is a divalent form of a C2-6 alkenyl, i.e., a C2- C 10 alkenylenyl. In one embodiment, the alkenylenyl is a divalent form of a C 2-14 alkenyl, i.e., a C2-C8 alkenylenyl. In one embodiment, the alkylenyl is a divalent form of an unsubstituted C2-6 alkenyl, i.e., a C2-C6 alkenylenyl. In another embodiment, the alkylenyl is a divalent form of an unsubstituted C 2-4 alkyl, i.e., a C 2 -C 4 alkenylenyl. Non-limiting exemplary alkenylenyl groups include -CH=CH-, -CH2CH=CH-, -CH2CH2CH=CHCH2-, and -CH 2 CH=CHCH 2 CH=CHCH 2 CH 2 -. [1035] The terms "alkoxyl" or "alkoxy" as used herein refers to an alkyl group, as defined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, and tert-butoxy. An "ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether is or resembles an alkoxyl, such as can be represented by one of -O-alkyl, -O-alkenyl, and -O- alkynyl. Aroxy can be represented by –O-aryl or O-heteroaryl, wherein aryl and heteroaryl are as defined below. The alkoxy and aroxy groups can be substituted as described above for alkyl. [1036] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, e.g., a moiety that can be represented by the general formula: wherein R9, R10, and R'10 each independently represent a hydrogen, an alkyl, an alkenyl, - (CH 2 ) m -R 8 or R 9 and R 10 taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure; R8 represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle or a polycycle; and m is zero or an integer in the range of 1 to 8. In some embodiments, only one of R9 or R10 can be a carbonyl, e.g., R9, R10 and the nitrogen together do not form an imide. In still other embodiments, the term "amine" does not encompass amides, e.g., wherein one of R9 and R10 represents a carbonyl. In additional embodiments, R 9 and R 10 (and optionally R' 10 ) each independently represent a hydrogen, an alkyl or cycloalkly, an alkenyl or cycloalkenyl, or alkynyl. Thus, the term "alkylamine" as used herein means an amine group, as defined above, having a substituted (as described above for alkyl) or unsubstituted alkyl attached thereto, i.e., at least one of R9 and R 10 is an alkyl group. [1037] The term "amido" is art-recognized as an amino-substituted carbonyl and includes a moiety that can be represented by the general formula: wherein R 9 and R 10 are as defined above. [1038] "Aryl", as used herein, refers to C5-C10-membered aromatic, heterocyclic, fused aromatic, fused heterocyclic, biaromatic, or bihetereocyclic ring systems. Broadly defined, "aryl", as used herein, includes 5-, 6-, 7-, 8-, 9-, and 10-membered single-ring aromatic groups that may include from zero to four heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "heteroaromatics". The aromatic ring can be substituted at one or more ring positions with one or more substituents including, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino (or quaternized amino), nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF 3 , -CN; and combinations thereof. [1039] The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (i.e., "fused rings") wherein at least one of the rings is aromatic, e.g., the other cyclic ring or rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocycles. Examples of heterocyclic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3 b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3- thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl and xanthenyl. One or more of the rings can be substituted as defined above for "aryl". [1040] The term "aralkyl," as used herein, refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group). [1041] The term "carbocycle," as used herein, refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon. [1042] "Heterocycle" or "heterocyclic," as used herein, refers to a cyclic radical attached via a ring carbon or nitrogen of a monocyclic or bicyclic ring containing 3-10 ring atoms, for example, from 5-6 ring atoms, consisting of carbon and one to four heteroatoms each selected from the group consisting of non-peroxide oxygen, sulfur, and N(Y) wherein Y is absent or is H, O, (C1-C10) alkyl, phenyl or benzyl, and optionally containing 1-3 double bonds and optionally substituted with one or more substituents. Examples of heterocyclic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxepanyl, oxetanyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H- pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4- thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl and xanthenyl. Heterocyclic groups can optionally be substituted with one or more substituents at one or more positions as defined above for alkyl and aryl, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic or heteroaromatic moiety, -CF3, and -CN. [1043] The term "carbonyl" is art-recognized and includes such moieties as can be represented by the general formula: wherein X is a bond or represents an oxygen or a sulfur, and R11 represents a hydrogen, an alkyl, a cycloalkyl, an alkenyl, a cycloalkenyl, or an alkynyl, R' 11 represents a hydrogen, an alkyl, a cycloalkyl, an alkenyl, a cycloalkenyl, or an alkynyl. Where X is an oxygen and R11 or R' 11 is not hydrogen, the formula represents an "ester". Where X is an oxygen and R 11 is as defined above, the moiety is referred to herein as a carboxyl group, and particularly when R11 is a hydrogen, the formula represents a "carboxylic acid". Where X is an oxygen and R' 11 is hydrogen, the formula represents a "formate". In general, where the oxygen atom of the above formula is replaced by sulfur, the formula represents a "thiocarbonyl" group. Where X is a sulfur and R 11 or R' 11 is not hydrogen, the formula represents a "thioester." Where X is a sulfur and R11 is hydrogen, the formula represents a "thiocarboxylic acid." Where X is a sulfur and R' 11 is hydrogen, the formula represents a "thioformate." On the other hand, where X is a bond, and R11 is not hydrogen, the above formula represents a "ketone" group. Where X is a bond, and R 11 is hydrogen, the above formula represents an "aldehyde" group. [1044] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Examples of heteroatoms are boron, nitrogen, oxygen, phosphorus, sulfur and selenium. Other useful heteroatoms include silicon and arsenic. [1045] As used herein, the term "nitro" means -NO 2 ; the term "halogen" designates -F, -Cl, - Br or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO 2 -. [1046] The term "substituted" as used herein, refers to all permissible substituents of the compounds described herein. In the broadest sense, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, but are not limited to, halogens, hydroxyl groups, or any other organic groupings containing any number of carbon atoms, for example, 1-14 carbon atoms, and optionally include one or more heteroatoms such as oxygen, sulfur, or nitrogen grouping in linear, branched, or cyclic structural formats. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aroxy, substituted aroxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C 3 -C 20 cyclic, substituted C3-C20 cyclic, heterocyclic, substituted heterocyclic, aminoacid, peptide, and polypeptide groups. [1047] As described herein, compounds of the present disclosure may contain "optionally substituted" moieties. In general, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable", as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein. [1048] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; —(CH 2 ) 0-4 R ∘ ; —(CH 2 ) 0-4 OR ∘ ; —O(CH 2 ) 0-4 R ∘ , —O—(CH2)0-4C(O)OR ∘ ; —(CH2)0-4CH(OR ∘ )2; —(CH2)0-4SR ∘ ; —(CH2)0-4Ph, which may be substituted with R ∘ ; —(CH2)0-4O(CH2)0-1Ph which may be substituted with R ∘ ; —CH═CHPh, which may be substituted with R ∘ ; —(CH2)0-4O(CH2)0-1-pyridyl which may be substituted with R ∘ ; —NO2; —CN; —N3; —(CH2)0-4N(R ∘ )2; —(CH2)0-4N(R ∘ )C(O)R ∘ ; —N(R ∘ )C(S)R ∘ ; — (CH 2 ) 0-4 N(R ∘ )C(O)NR ∘ 2 ; —N(R ∘ )C(S)NR ∘ 2 ; —(CH 2 ) 0-4 N(R ∘ )C(O)OR ∘ ; — N(R ∘ )N(R ∘ )C(O)R ∘ ; —N(R ∘ )N(R ∘ )C(O)NR ∘ 2 ; —N(R ∘ )N(R ∘ )C(O)OR ∘ ; —(CH 2 ) 0-4 C(O)R ∘ ; — C(S)R ∘ ; —(CH 2 ) 0-4 C(O)OR ∘ ; —(CH 2 ) 0-4 C(O)SR ∘ ; —(CH 2 ) 0-4 C(O)OSiR ∘ 3 ; —(CH 2 ) 0- 4OC(O)R ∘ ; —OC(O)(CH2)0-4SR ∘ , SC(S)SR ∘ ; —(CH2)0-4SC(O)R ∘ ; —(CH2)0-4C(O)NR ∘ 2; — C(S)NR ∘ 2; —C(S)SR ∘ ; —SC(S)SR ∘ , —(CH2)0-4OC(O)NR ∘ 2; —C(O)N(OR ∘ )R ∘ ; — C(O)C(O)R ∘ ; —C(O)CH2C(O)R ∘ ; —C(NOR ∘ )R ∘ ; —(CH2)0-4SSR ∘ ; —(CH2)0-4S(O)2R ∘ ; — (CH 2 ) 0-4 S(O) 2 OR ∘ ; —(CH 2 ) 0-4 OS(O) 2 R ∘ ; —S(O) 2 NR ∘ 2 ; —(CH 2 ) 0-4 S(O)R ∘ ; — N(R ∘ )S(O) 2 NR ∘ 2 ; —N(R ∘ )S(O) 2 R ∘ ; —N(OR ∘ )R ∘ ; —C(NH)NR ∘ 2 ; —P(O) 2 R ∘ ; —P(O)R ∘ 2 ; — OP(O)R ∘ 2; —OP(O)(OR ∘ )2; SiR ∘ 3; —(C1-4 straight or branched alkylene)O—N(R ∘ )2; or — (C1-4 straight or branched alkylene)C(O)O—N(R ∘ )2, wherein each R ∘ may be substituted as defined below and is independently hydrogen, C1-6 aliphatic, —CH2Ph, —O(CH2)0-1Ph, — CH2-(5-6 membered heteroaryl ring), or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R ∘ , taken together with their intervening atom(s), form a 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below. [1049] Suitable monovalent substituents on R ∘ (or the ring formed by taking two independent occurrences of R ∘ together with their intervening atoms), are independently halogen, —(CH 2 ) 0- 2R ● , -(haloR ● ), —(CH2)0-2OH, —(CH2)0-2OR ● , —(CH2)0-2CH(OR ● )2; —O(haloR ● ), —CN, — N 3 , —(CH 2 ) 0-2 C(O)R ● , —(CH 2 ) 0-2 C(O)OH, —(CH 2 ) 0-2 C(O)OR ● , —(CH 2 ) 0-2 SR ● , —(CH 2 ) 0- 2SH, —(CH2)0-2NH2, —(CH2)0-2NHR ● , —(CH2)0-2NR ● 2, —NO2, —SiR ● 3, —OSiR ● 3, — C(O)SR ● , —(C 1-4 straight or branched alkylene)C(O)OR ● , or —SSR ● wherein each R ● is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently selected from C 1-4 aliphatic, —CH 2 Ph, —O(CH 2 ) 0-1 Ph, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R ∘ include ═O and ═S. [1050] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: ═O, ═S, ═NNR* 2 , ═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O) 2 R*, ═NR*, ═NOR*, —O(C(R* 2 )) 2-3 O—, or —S(C(R* 2 )) 2-3 S—, wherein each independent occurrence of R* is selected from hydrogen, C1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents that are bound to vicinal substitutable carbons of an "optionally substituted" group include: —O(CR*2)2-3O—, wherein each independent occurrence of R* is selected from hydrogen, C 1-6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [1051] Suitable substituents on the aliphatic group of R* include halogen, —R ● , -(haloR ● ), — OH, —OR ● , —O(haloR ● ), —CN, —C(O)OH, —C(O)OR ● , —NH 2 , —NHR ● , —NR ● 2 , or — NO2, wherein each R ● is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently C 1-4 aliphatic, —CH 2 Ph, —O(CH 2 ) 0-1 Ph, or a 5-6- membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [1052] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include —R † , —NR † 2 , —C(O)R † , —C(O)OR † , —C(O)C(O)R † , —C(O)CH 2 C(O)R † , — S(O)2R † , —S(O)2NR † 2, —C(S)NR † 2, —C(NH)NR † 2, or —N(R † )S(O)2R † ; wherein each R † is independently hydrogen, C 1-6 aliphatic which may be substituted as defined below, unsubstituted —OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of R † , taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [1053] Suitable substituents on the aliphatic group of R † are independently halogen, —R ● , - (haloR ● ), —OH, —OR ● , —O(haloR ● ), —CN, —C(O)OH, —C(O)OR ● , —NH2, —NHR ● , — NR ● 2 , or —NO 2 , wherein each R ● is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently C1-4 aliphatic, —CH2Ph, —O(CH2)0-1Ph, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. [1054] Heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. It is understood that "substitution" or "substituted" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation, for example, by rearrangement, cyclization, or elimination. [1055] In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein. The permissible substituents can be one or more and the same or different for appropriate organic compounds. The heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. [1056] In various embodiments, the substituent is selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, each of which optionally is substituted with one or more suitable substituents. In some embodiments, the substituent is selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, wherein each of the alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone can be further substituted with one or more suitable substituents. [1057] Examples of substituents include, but are not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, thioketone, ester, heterocyclyl, –CN, aryl, aryloxy, perhaloalkoxy, aralkoxy, heteroaryl, heteroaryloxy, heteroarylalkyl, heteroaralkoxy, azido, alkylthio, oxo, acylalkyl, carboxy esters, carboxamido, acyloxy, aminoalkyl, alkylaminoaryl, alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylamino, alkylsulfonyl, carboxamidoalkylaryl, carboxamidoaryl, hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxy, aminocarboxamidoalkyl, cyano, alkoxyalkyl, perhaloalkyl, arylalkyloxyalkyl, and the like. In some embodiments, the substituent is selected from cyano, halogen, hydroxyl, and nitro. [1058] Antibodies: As used herein, the term "antibody" is referred to in the broadest sense and specifically covers various embodiments including, but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies formed from at least two intact antibodies), and antibody fragments (e.g., diabodies) so long as they exhibit a desired biological activity (e.g., "functional"). Antibodies are primarily amino-acid based molecules but may also comprise one or more modifications (including, but not limited to the addition of sugar moieties, fluorescent moieties, chemical tags, etc.). Non-limiting examples of antibodies or fragments thereof include VH and VL domains, scFvs, Fab, Fab', F(ab')2, Fv fragment, diabodies, linear antibodies, single chain antibody molecules, multispecific antibodies, bispecific antibodies, intrabodies, monoclonal antibodies, polyclonal antibodies, humanized antibodies, codon-optimized antibodies, tandem scFv antibodies, bispecific T-cell engagers, mAb2 antibodies, chimeric antigen receptors (CAR), tetravalent bispecific antibodies, biosynthetic antibodies, native antibodies, miniaturized antibodies, unibodies, maxibodies, antibodies to senescent cells, antibodies to conformers, antibodies to disease specific epitopes, or antibodies to innate defense molecules. [1059] Associated: As used herein, the terms "associated with," "conjugated," "linked," "attached," and "tethered," when used with respect to two or more moieties, means that the moieties are physically associated or connected with one another, either directly or via one or more additional moieties that serves as a linking agent, to form a structure that is sufficiently stable so that the moieties remain physically associated under the conditions in which the structure is used, e.g., physiological conditions. An "association" need not be strictly through direct covalent chemical bonding. It may also suggest ionic or hydrogen bonding or a hybridization based connectivity sufficiently stable such that the "associated" entities remain physically associated. [1060] Cargo: As used herein, the term "cargo" or "payload" can refer to one or more molecules or structures encompassed in a delivery vehicle for delivery to or into a cell or tissue. Non-limiting examples of cargo can include a nucleic acid, a polypeptide, a peptide, a protein, a liposome, a label, a tag, a small chemical molecule, a large biological molecule, and any combinations thereof. [1061] Chimeric Antigen Receptors (CARs): As used herein, the term "chimeric antigen receptor" or "CAR" refers to an artificial chimeric protein comprising at least one antigen specific targeting region (ASTR), a transmembrane domain and an intracellular signaling domain, wherein the antigen specific targeting region comprises a full-length antibody or a fragment thereof. Any molecule that is capable of binding a target antigen with high affinity can be used in the ASTR of a CAR. The CAR may optionally have an extracellular spacer domain and/or a co-stimulatory domain. A CAR may also be used to generate a cytotoxic cell carrying the CAR. [1062] Circular RNA: As used herein, the term "circular RNA" or "circRNA" refer to a RNA that forms a circular structure through covalent or non-covalent bonds. [1063] Co-Administered: As used herein, the term "co-administered" or "co-administering" means administering an originator construct, a benchmark construct or a targeting system with one or more additional an originator construct, a benchmark construct, a targeting systems or other therapeutic agents or moieties sufficiently close in time such that the effect of the originator construct, a benchmark construct, a targeting systems or other therapeutic agents or moieties is enhanced. [1064] Complementary and substantially complementary: As used herein, the term "complementary" refers to the ability of polynucleotides to form base pairs with one another. Base pairs are typically formed by hydrogen bonds between nucleotide units in antiparallel polynucleotide strands. Complementary polynucleotide strands can form base pair in the Watson-Crick manner (e.g., A to T, A to U, C to G), or in any other manner that allows for the formation of duplexes. As persons skilled in the art are aware, when using RNA as opposed to DNA, uracil rather than thymine is the base that is considered to be complementary to adenosine. However, when a U is denoted in the context of the present disclosure, the ability to substitute a T is implied, unless otherwise stated. Perfect complementarity or 100% complementarity refers to the situation in which each nucleotide unit of one polynucleotide strand can form hydrogen bond with a nucleotide unit of a second polynucleotide strand. Less than perfect complementarity refers to the situation in which some, but not all, nucleotide units of two strands can form hydrogen bond with each other. For example, for two 20-mers, if only two base pairs on each strand can form hydrogen bond with each other, the polynucleotide strands exhibit 10% complementarity. In the same example, if 18 base pairs on each strand can form hydrogen bonds with each other, the polynucleotide strands exhibit 90% complementarity. As used herein, the term "substantially complementary" means that the siRNA has a sequence (e.g., in the antisense strand) which is sufficient to bind the desired target mRNA, and to trigger the RNA silencing of the target mRNA. [1065] Delivery: As used herein, "delivery" refers to the act or manner of delivering a compound, substance, entity, moiety, cargo or payload. [1066] DNA and RNA: As used herein, the term "RNA" or "RNA molecule" or "ribonucleic acid molecule" refers to a polymer of ribonucleotides; the term "DNA" or "DNA molecule" or "deoxyribonucleic acid molecule" refers to a polymer of deoxyribonucleotides. DNA and RNA can be synthesized naturally, e.g., by DNA replication and transcription of DNA, respectively; or be chemically synthesized. DNA and RNA can be single-stranded (i.e., ssRNA or ssDNA, respectively) or multi-stranded (e.g., double stranded, i.e., dsRNA and dsDNA, respectively). The term "mRNA" or "messenger RNA", as used herein, refers to a single stranded RNA that encodes the amino acid sequence of one or more polypeptide chains. [1067] Encapsulate: As used herein, the term "encapsulate" means to enclose, surround or encase. [1068] Encode: As used herein the term "encode" refers broadly to any process whereby the information in a polymeric macromolecule is used to direct the production of a second molecule that is different from the first. The second molecule may have a chemical structure that is different from the chemical nature of the first molecule. [1069] Enhance expression of a gene: As used herein, the phrase "add-back" or "enhance expression of a gene" means to cause an increase in the amount of an expression product of the gene. The expression product can be an RNA transcribed from the gene (e.g., an mRNA) or a polypeptide translated from an mRNA transcribed from the gene. Typically, an increase in the level of an mRNA results in an increase in the level of a polypeptide translated therefrom. The level of expression may be determined using standard techniques for measuring mRNA or protein. [1070] Exosomes: As used herein, "exosome" is a vesicle secreted by mammalian cells or a complex involved in RNA degradation. [1071] Formulation: As used herein, a "formulation" includes at least one compound, substance, entity, moiety, cargo or payload and a delivery agent. [1072] Fragment: A "fragment," as used herein, refers to a portion. For example, fragments of proteins may comprise polypeptides obtained by digesting full-length protein isolated from cultured cells. [1073] Homology: As used herein, the term "homology" refers to the overall relatedness between polymeric molecules, e.g. between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical or similar. The term "homologous" necessarily refers to a comparison between at least two sequences (polynucleotide or polypeptide sequences). In accordance with the disclosure, two polynucleotide sequences are considered to be homologous if the polypeptides they encode are at least about 50%, 60%, 70%, 80%, 90%, 95%, or even 99% for at least one stretch of at least about 20 amino acids. In some embodiments, homologous polynucleotide sequences are characterized by the ability to encode a stretch of at least 4–5 uniquely specified amino acids. For polynucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode a stretch of at least 4–5 uniquely specified amino acids. In accordance with the disclosure, two protein sequences are considered to be homologous if the proteins are at least about 50%, 60%, 70%, 80%, or 90% identical for at least one stretch of at least about 20 amino acids. [1074] Inactive Ingredient: As used herein, the term "inactive ingredient" refers to one or more agents that do not contribute to the activity of the active ingredient of the pharmaceutical composition included in formulations. In some embodiments, all, none or some of the inactive ingredients which may be used in the formulations of the present disclosure may be approved by the US Food and Drug Administration (FDA). [1075] IRES: As used herein, the term "internal ribosome entry site" or "IRES" refers to an RNA sequence or structural element ranging in size form 10 nucleotides to 1,000 nucleotides or more which is capable of initiating translation of a polypeptide in the absence of a normal RNA cap structure. [1076] Identity: As used herein, the term "identity" refers to the overall relatedness between polymeric molecules, e.g., between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Calculation of the percent identity of two polynucleotide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using methods such as those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; each of which is incorporated herein by reference. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4:11-17), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. Methods commonly employed to determine percent identity between sequences include, but are not limited to those disclosed in Carillo, H. and Lipman, D., SIAM J Applied Math., 48:1073 (1988); incorporated herein by reference. Techniques for determining identity are codified in publicly available computer programs. Exemplary computer software to determine homology between two sequences include, but are not limited to, GCG program package, Devereux, J., et al., Nucleic Acids Research, 12(1), 387 (1984)), BLASTP, BLASTN, and FASTA Altschul, S. F. et al., J. Molec. Biol., 215, 403 (1990)). [1077] Inhibit expression of a gene: As used herein, the phrase "knock-down" or "inhibit expression of a gene" means to cause a reduction in the amount of an expression product of the gene. The expression product can be an RNA transcribed from the gene (e.g., an mRNA) or a polypeptide translated from an mRNA transcribed from the gene. Typically, a reduction in the level of an mRNA results in a reduction in the level of a polypeptide translated therefrom. The level of expression may be determined using standard techniques for measuring mRNA or protein. [1078] Ionizable Lipid: As used herein "ionizable lipid" refers to any of a number of lipid species that carry a net positive charge at a selected pH. [1079] Lipid Nanoparticle: As used herein "lipid nanoparticle" or "LNP" refers to a delivery vehicle comprising one or more lipids (e.g., cationic lipids, non-cationic lipids, PEG-modified lipids). [1080] Liposome: As used herein "liposome" generally refers to a vesicle composed of lipids (e.g., amphiphilic lipids) arranged in one or more spherical bilayers or bilayers. [1081] Modified: As used herein "modified" refers to a changed state or structure of a molecule. Molecules may be modified in many ways including chemically, structurally, and functionally. [1082] Non-Cationic Lipid: As used herein "non-cationic lipid" refers to any neutral, zwitterionic or anionic lipid. [1083] Peptide: As used herein, "peptide" is less than or equal to 50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids long. [1084] Pharmaceutical Composition: As used herein the term "pharmaceutical composition" refers to compositions comprising at least one active ingredient and optionally one or more pharmaceutically acceptable excipients. [1085] PEG: As used herein "PEG" means any polyethylene glycol or other polyalkylene ether polymer. [1086] Spacer: As used herein the term "spacer" refers to a region of a polynucleotide or polypeptide ranging from 1 residue to hundreds or thousands of residues separating two other elements in a sequence. The sequence of the spacer can be defined or random. A spacer sequence is typically non-coding but may be a coding sequence. [1087] Sterol: As used herein "sterol" is a subgroup of steroids consisting of steroid alcohols. [1088] Structural Lipid: As used herein "structural lipid" refers to sterols and lipids containing sterol moieties. [1089] Transcription: As used herein the term "transcription" refers to the formation or synthesis of an RNA molecule by an RNA polymerase using a DNA molecule as a template. [1090] Translation: As used herein the term "translation" refers to the formation of a polypeptide molecule by a ribosome based upon a RNA template. [1091] Treat and Prevent: As used herein the terms "treat" or "prevent" as well as words stemming therefrom do not necessarily imply 100% or complete treatment or prevention. Rather there are varying degrees of treatment or prevention of which one of ordinary skill in the art recognizes as having a potential benefit or therapeutic effect. Also, "prevention" can encompass delaying the onset of the disease, symptom or condition thereof. [1092] Unmodified: As used herein, "unmodified" refers to any substance, compound or molecule prior to being changed in any way. Unmodified may, but does not always, refer to the wild type or native form of a biomolecule. Molecules may undergo a series of modifications whereby each modified molecule may serve as the "unmodified" starting molecule for a subsequent modification. [1093] Vector: As used herein, a "vector" is any molecule or moiety which transports, transduces or otherwise acts as a carrier of a heterologous molecule. Vectors of the present disclosure may be produced recombinantly and may be based on and/or may comprise viral parent or reference sequence. Such parent or reference viral sequences may serve as an original, second, third or subsequent sequence for engineering vectors. In non-limiting examples, such parent or reference viral sequences may comprise any one or more of the following sequences: a polynucleotide sequence encoding a polypeptide or multi-polypeptide, which sequence may be wild-type or modified from wild-type and which sequence may encode full-length or partial sequence of a protein, protein domain, or one or more subunits of a protein; a polynucleotide comprising a modulatory or regulatory nucleic acid which sequence may be wild-type or modified from wild-type; and a transgene that may or may not be modified from wild-type sequence . These viral sequences may serve as either the "donor" sequence of one or more codons (at the nucleic acid level) or amino acids (at the polypeptide level) or "acceptor" sequences of one or more codons (at the nucleic acid level) or amino acids (at the polypeptide level).The details of one or more embodiments of the disclosure are set forth in the accompanying description below. Although any materials and methods similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred materials and methods are now described. Other features, objects and advantages of the disclosure will be apparent from the description. In the description, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the case of conflict, the present description will control. [1094] XI. PARTICULAR EMBODIMENTS [1095] Embodiment 1. A compound having the structure of Formula (CY-I): [1096] [1097] or a pharmaceutically acceptable salt thereof, wherein: [1098] R 1 is -OH, [1099] [1100] wherein Z 1 is optionally substituted C1-C6 alkyl; [1101] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1102] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1103] X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; [1104] Y 1 and Y 2 are independently [1105] R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl. [1106] Embodiment 2. A compound having the structure of Formula (CY-II): , [1108] or a pharmaceutically acceptable salt thereof, wherein: [1109] R1 is -OH, , [1110] wherein Z 1 is optionally substituted C1-C6 alkyl; [1111] X 1 is optionally substituted C2-C6 alkylenyl; [1112] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1113] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1114] Y 1 and Y 2 are independently [1115] R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl. [1116] Embodiment 3. A compound having the structure of Formula (CY-III): , [1118] or a pharmaceutically acceptable salt thereof, wherein: [1119] R 1 is -OH, , [1120] wherein Z 1 is optionally substituted C1-C6 alkyl; [1121] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1122] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1123] X 4 and X 5 are independently optionally substituted C 2 -C 14 alkylenyl; [1124] Y 1 and Y 2 are independently [1125] R 2 and R 3 are independently optionally substituted C4-C20 alkyl. [1126] Embodiment 4. A compound having the structure of Formula (CY-IV): , [1128] or a pharmaceutically acceptable salt thereof, wherein: [1129] R 1 is -OH, , [1130] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1131] X 1 is optionally substituted C2-C6 alkylenyl; [1132] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1133] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1134] Y 1 and Y 2 are independently [1135] R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl. [1136] Embodiment 5. A compound having the structure of Formula (CY-I): , [1138] or a pharmaceutically acceptable salt thereof, wherein: [1139] R 1 is -OH, R 1a , , [1141] wherein Z 1 is optionally substituted C1-C6 alkyl; [1142] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1143] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1144] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1145] Y 1 and Y 2 are independently ; [1147] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1148] R 1a is: [1154] Embodiment 6. The compound of Embodiment 5, wherein R 1 is -OH, : [1158] or a pharmaceutically acceptable salt thereof, wherein: [1159] R 1 is -OH, R 1a , , [1161] wherein Z 1 is optionally substituted C1-C6 alkyl; [1162] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1163] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1164] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1165] Y 1 and Y 2 are independently ; [1167] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1168] R 1a is: [1174] Embodiment 8. The compound of Embodiment 7, wherein R 1 is -OH, : [1178] or a pharmaceutically acceptable salt thereof, wherein: [1179] R 1 is -OH, R 1a , , [1181] wherein Z 1 is optionally substituted C1-C6 alkyl; [1182] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1183] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1184] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1185] Y 1 and Y 2 are independently ; [1187] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1188] R 1a is: ; [1194] Embodiment 10. The compound of Embodiment 9, wherein R 1 is -OH, : [1198] or a pharmaceutically acceptable salt thereof, wherein: [1199] R 1 is -OH, R 1a , , [1201] wherein Z 1 is optionally substituted C1-C6 alkyl; [1202] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1203] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1204] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1205] Y 1 and Y 2 are independently ; [1207] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1208] R 1a is: [1214] Embodiment 12. The compound of Embodiment 11, wherein R 1 is -OH, : [1218] or a pharmaceutically acceptable salt thereof, wherein: [1219] R 1 is -OH, R 1a , , [1221] wherein Z 1 is optionally substituted C1-C6 alkyl; [1222] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1223] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1224] Y 1 and Y 2 are independently ; [1226] X 6 and X 7 are independently -CH 2 - or -CH 2 CH 2 -; [1227] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1228] R 1a is: ; [1234] Embodiment 14. The compound of Embodiment 13, wherein R 1 is -OH, : [1238] or a pharmaceutically acceptable salt thereof, wherein: [1239] R 1 is -OH, R 1a , ; [1241] Z 1 is optionally substituted C1-C6 alkyl; [1242] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1243] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1244] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl or optionally substituted C 2 -C 14 alkenylenyl; [1245] Y 1 and Y 2 are independently , [1247] wherein the bond marked with an "*" is attached to X 4 or X 5 ; [1248] each Z 2 is independently H or optionally substituted C 1 -C 8 alkyl; [1249] each Z 3 is indpendently optionally substituted C1-C6 alkylenyl; [1250] R 2 is optionally substituted C 4 -C 20 alkyl, optionally substituted C 2 -C 14 alkenylenyl, or -CH(OR 6 )(OR 7 ); [1251] R 3 is optionally substituted C4-C20 alkyl, optionally substituted C2-C14 alkenylenyl, or -CH(OR 8 )(OR 9 ); [1252] R 1a is: ; 1-C14 alkyl, optionally substituted C 2 -C 14 alkenylenyl, or -(CH 2 ) m -A-(CH 2 ) n H; [1259] A is a C3-C8 cycloalkylenyl; [1260] each m is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; and [1261] each n is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. [1262] Embodiment 16. The compound of Embodiment 15, wherein: [1263] R 1 is -OH, R 1a , , [1265] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1266] X 1 is optionally substituted C2-C6 alkylenyl; [1267] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1268] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1269] Y 1 and Y 2 are independently ; [1271] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1272] R 1a is: [1278] Embodiment 17. The compound of Embodiment 16, wherein: [1279] R 1 is -OH, R 1a , , [1281] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1282] X 1 is optionally substituted C2-C6 alkylenyl; [1283] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1284] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1285] Y 1 and Y 2 are independently ; [1287] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1288] R 1a is: [1294] Embodiment 18. The compound of Embodiments 16 or 17, wherein R 1 is -OH, . [1296] Embodiment 19. The compound of Embodiments 16 or 17, wherein Y 1 and Y 2 are independently: [1297] . [1298] Embodiment 20. The compound of any one of Embodiments 15, 18, or 19, wherein R 2 is -CH(OR 6 )(OR 7 ). [1299] Embodiment 21. The compound of any one of Embodiments 15 or 18-20, wherein R 3 is -CH(OR 8 )(OR 9 ). [1300] Embodiment 22. A compound having the structure of Formula (CY-II): , [1302] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I) of Embodiment 15. [1303] Embodiment 23. The compound of Embodiment 22, wherein: [1304] R 1 is -OH, R 1a , , [1306] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1307] X 1 is optionally substituted C2-C6 alkylenyl; [1308] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1309] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1310] Y 1 and Y 2 are independently ; [1312] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1313] R 1a is: [1319] Embodiment 24. The compound of Embodiments 22 or 23, wherein R 1 is -OH, . [1321] Embodiment 25. The compound of Embodiments 22 or 23, wherein Y 1 and Y 2 are independently: . [1323] Embodiment 26. The compound of any one of Embodiments 22, 24 or 25, wherein R 2 is -CH(OR 6 )(OR 7 ). [1324] Embodiment 27. The compound of any one of Embodiments 22 or 24-26, wherein R 3 is -CH(OR 8 )(OR 9 ). [1325] Embodiment 28. A compound having the structure of Formula (CY-III): , [1327] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I) in Embodiment 15. [1328] Embodiment 29. The compound of Embodiment 28, wherein: [1329] R 1 is -OH, R 1a , , [1331] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1332] X 1 is optionally substituted C2-C6 alkylenyl; [1333] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1334] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1335] Y 1 and Y 2 are independently ; [1337] R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl; [1338] R 1a is: [1343] R 5a , R 5b , and R 5c are independently hydrogen and C1-C6 alkyl. [1344] Embodiment 30. The compound of Embodiments 28 or 29, wherein R 1 is -OH, . [1346] Embodiment 31. The compound of Embodiments 28 or 30, wherein Y 1 and Y 2 are independently: . [1348] Embodiment 32. The compound of any one of Embodiments 28, 30, or 31, wherein R 2 is -CH(OR 6 )(OR 7 ). [1349] Embodiment 33. The compound of any one of Embodiments 28 or 30-32, wherein R 3 is -CH(OR 8 )(OR 9 ). [1350] Embodiment 34. A compound having the structure of Formula (CY-IV): , [1352] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I) of Embodiment 15. [1353] Embodiment 35. The compound of Embodiment 34, wherein: [1354] R 1 is -OH, R 1a , , [1356] wherein Z 1 is optionally substituted C 1 -C 6 alkyl; [1357] X 1 is optionally substituted C2-C6 alkylenyl; [1358] X 2 and X 3 are independently a bond, -CH 2 -, or -CH 2 CH 2 -; [1359] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1360] Y 1 and Y 2 are independently ; [1362] R 2 and R 3 are independently optionally substituted C 4 -C 20 alkyl; [1363] R 1a is: [1369] Embodiment 36. The compound of Embodiments 34 or 35, wherein R 1 is -OH, . [1371] Embodiment 37. The compound of Embodiments 34 or 36, wherein Y 1 and Y 2 are independently: . [1373] Embodiment 38. The compound of any one of Embodiments 34, 36, or 37, wherein R 2 is -CH(OR 6 )(OR 7 ). [1374] Embodiment 39. The compound of any one of Embodiments 34 or 36-38, wherein R 3 is -CH(OR 8 )(OR 9 ). [1375] Embodiment 40. A compound having the structure of Formula (CY-V): , [1377] or a pharmaceutically acceptable salt thereof, wherein X 6 and X 7 are independently - CH2- or -CH2CH2-; and R 1 , R 2 , R 3 , X 1 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I) of Embodiment 15. [1378] Embodiment 41. The compound of Embodiment 40, wherein: [1379] R 1 is -OH, R 1a , , [1381] wherein Z 1 is optionally substituted C1-C6 alkyl; [1382] X 1 is optionally substituted C 2 -C 6 alkylenyl; [1383] X 2 and X 3 are independently a bond, -CH2-, or -CH2CH2-; [1384] X 4 and X 5 are independently optionally substituted C2-C14 alkylenyl; [1385] Y 1 and Y 2 are independently ; [1387] R 2 and R 3 are independently optionally substituted C4-C20 alkyl; [1388] R 1a is: [1394] Embodiment 42. The compound of Embodiments 40 or 41, wherein R 1 is -OH, . [1396] Embodiment 43. The compound of Embodiments 40 or 41, wherein Y 1 and Y 2 are independently: [1397] . [1398] Embodiment 44. The compound of any one of Embodiments 40, 42 or 43, wherein R 2 is -CH(OR 6 )(OR 7 ). [1399] Embodiment 45. The compound of any one of Embodiments 40 or 42-44, wherein R 3 is -CH(OR 8 )(OR 9 ). [1400] Embodiment 46. A compound having the structure of Formula (CY-VI): , [1402] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 6 , R 7 , R 8 , R 9 , X 1 , X 2 , X 4 , X 5 , Y 1 , and Y 2 are as defined in connection with Formula (CY-I) of Embodiment 15. [1403] Embodiment 47. The compound of Embodiment 46, or a pharmaceutically acceptable salt thereof, wherein R 1 is -OH. [1404] Embodiment 48. The compound of Embodiments 46 or 47, or a pharmaceutically acceptable salt thereof, wherein X 1 is C2-C6 alkylenyl. [1405] Embodiment 49. The compound of any one of Embodiments 46-48, or a pharmaceutically acceptable salt thereof, wherein X 2 is -CH2CH2-. [1406] Embodiment 50. The compound of any one of Embodiments 46-49, or a pharmaceutically acceptable salt thereof, wherein X 4 is C2-C6 alkylenyl. [1407] Embodiment 51. The compound of any one of Embodiments 46-50, or pharmaceutically acceptable salt thereof, wherein X 5 is C 2 -C 6 alkylenyl. [1408] Embodiment 52. The compound of any one of Embodiments 46-41, or a pharmaceutically acceptable salt thereof, wherein Y 1 is: . [1410] Embodiment 53. The compound of any one of Embodiments 46-52, or a pharmaceutically acceptable salt thereof, wherein Y 2 is: . [1412] Embodiment 54. The compound of any one of Embodiments 46-53, or a pharmaceutically acceptable salt thereof, wherein each Z 3 is independently optionally substituted C1-C6 alkylenyl. [1413] Embodiment 55. The compound of any one Embodiments 46-54, or a pharmaceutically acceptable salt thereof, wherein each Z 3 is -CH2CH2-. [1414] Embodiment 56. The compound of any one of Embodiments 46-55, or a pharmaceutically acceptable salt thereof, wherein R 6 is C5-C14 alkyl. [1415] Embodiment 57. The compound of any one of Embodiments 46-56, or a pharmaceutically acceptable salt thereof, wherein R 7 is C 6 -C 14 alkyl. [1416] Embodiment 58. The compound of any one of Embodiments 46-55 or 57, or a pharmaceutically acceptable salt thereof, wherein R 6 is C 5 -C 14 alkenyl. [1417] Embodiment 59. The compound of any one of Embodiments 46-56 or 58, or a pharmaceutically acceptable salt thereof, wherein R 7 is C 6 -C 14 alkenyl. [1418] Embodiment 60. The compound of any one of Embodiments 46-59, or a pharmaceutically acceptable salt thereof, wherein R 8 is C 5 -C 16 alkyl. [1419] Embodiment 61. The compound of any one of Embodiments 46-60, or a pharmaceutically acceptable salt thereof, wherein R 9 is C 6 -C 14 alkyl. [1420] Embodiment 62. The compound of any one of Embodiments 46-59 or 61, or a pharmaceutically acceptable salt thereof, wherein R 8 is C 5 -C 16 alkenyl. [1421] Embodiment 63. The compound of any one of Embodiments 46-60 or 62, or a pharmaceutically acceptable salt thereof, wherein R 9 is C 6 -C 14 alkyl. [1422] The present disclosure is further illustrated by the following non-limiting examples. XII. EXAMPLES Methods of Making the Lipids [1423] The Lipids of the Disclosure may be prepared using any convenient methodology. In a rational approach, the lipids are constructed from their individual components. The components can be covalently bonded to one another through functional groups, as is known in the art, where such functional groups may be present on the components or introduced onto the components using one or more steps, e.g., oxidation reactions, reduction reactions, cleavage reactions and the like. Functional groups that may be used in covalently bonding the components together to produce the lipids: hydroxy, sulfhydryl, amino, and the like. Where necessary and/or desired, certain moieties on the components may be protected using blocking groups, as is known in the art, see, e.g., Green & Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons) (1991). [1424] Alternatively, the lipids can be produced using known combinatorial methods to produce large libraries of potential lipids which may then be screened for identification of a lipid with desired functionalities. Methods of Making the Delivery Vehicles [1425] The delivery vehicles such as LNPs of the present disclosure may be prepared using any convenient methodology. In one non-limiting example, the LNPs are formed by mixing equal volumes of lipids dissolved in alcohol with oligonucleotide payloads dissolved in a citrate buffer by an impinging jet process. [1426] The lipid solution contains a cationic lipid compound of the present disclosure, a helper lipid, a neutral lipid and a PEGylated lipid. The payload to total lipid ratio is approximately 1:20 (wt/wt). The LNPs are formed by mixing equal volumes of lipid solution in ethanol with oligonucleotide payloads dissolved in a citrate buffer by an impinging jet process through a mixing device. The mixed LNP solution is held at room temperature for 0-24 hrs prior to a dilution step. [1427] The solution is then concentrated and diafiltered with suitable buffer by ultrafiltration or dialysis process using membranes. The final product is sterile filtered and stored at 4° C. Evaluation of Candidate LNP Targeting Systems [1428] A library of candidate targeting systems is prepared where the candidate targeting systems comprise at least one identifier sequence or moiety in the formulation and at least one identifier sequence and/or payload in the nucleic acid construct. Candidate Targeting System Generation [1429] A population of lipid nanoparticle (LNP) formulations are generated where the cationic lipid component is labeled with at least one identifier sequence or moiety. The LNP formulations that are generated may include LNPs where (a) the components are the same for all formulations and the molar ratios of the components are the same for all the LNP formulations, (b) the components are the same for all formulations but the molar ratios of the components are different for all the LNP formulations, or (c) the components are different for the LNP formulations. Each of the different LNP formulation can include different identifier sequence or moiety in order to track targeting system after administration. Nucleic acid constructs including at least one identifier sequence or payload (e.g., a reporter gene) is generated and formulated in the population of LNPs in order to create candidate targeting systems to be administered to a subject. Screening and Validation of Candidate Targeting Systems [1430] The candidate targeting systems are then administered into a subject at a pre-determined dose and dosing interval. After administration the entire subject or a region of the subject is screened to determine the location of the LNP formation and/or the payload of the benchmark construct. The subject can be scanned by various methods known in the art including positron emission tomography (PET) and computed tomography (CT) utilizing the 64 Cu radiolabel. The localization of the LNP formation and/or the payload the will be determined by visual inspection of the PET images for areas with the greatest concentration of 64 Cu and anatomical position of PET results will be confirmed using the results of the CT scan. The scan can be repeated in order to determine if the localization changes over time. [1431] At the desired time points, samples will be taken from the areas of the subject displaying localization of the LNP and/or the payload in the whole animal localization screening performed above for higher resolution screening of the distribution. [1432] The samples can then be prepared for Fluorescence-activated Cell Sorting (FACS) via the directions supplied with the cell sorter. These populations of cells are then prepared for deep sequencing to determine the presence and identity of the payload and/or identifier sequence. [1433] These results from the screening of the LNP library will provide the tropism of the LNP formulation and the nucleic acid construct that was administered to the subject. Synthesis of exemplary ionizable lipids Synthesis of Select Intermediates [1434] Synthesis of ethyl 3-pentyloct-2-enoate (L1-2) [1435] To a solution of triethyl phophonoacetate (26.3 g, 118 mmol) in anhydrous THF (33 mL) was added dropwise 1M NaHMDS in THF (118 mL, 118 mmol) at -10 to -15 °C under nitrogen atmosphere. After completion of addition, the mixture was stirred at -10 to -15 °C for 30 min and then at 0 °C for 1h. To this mixture was dropped in 6-undecanone (10.0 g, 59 mmol) at 0 °C and the reaction mixture was allowed to warm to room temperature and stirred overnight. The mixture was then warmed to 45 °C and stirred for 24h. Aq. sat. NH 4 Cl (8 mL) was added and the THF was evaporated. The residue was mixed with Et2O (80 mL) and H2O (100 mL) and the resulting phases were separated. The aqueous phase was extracted with Et2O (80 mL). Combined organic phases were washed with H 2 O (100 mL x 2) and dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO 2 : 0 to 4% ethyl acetate in hexane gradient) to yield ethyl 3-pentyloct-2-enoate L1-2 as colorless oil (11.6 g, 82%). 1 H-NMR (300 MHz, CDCl3) δ 5.60 (s, 1H), 4.13 (q, J = 7.1 Hz, 2H), 2.57 (t, J = 7.6 Hz, 2H), 2.12 (t, J = 7.4 Hz, 2H), 1.52-1.19 (m, 15H), 0.89 (t, J = 7.2 Hz, 6H); CIMS m/z 241 [M+H] + . [1436] Synthesis of ethyl 3-pentyloctanoate (L1-3) [1437] To a solution of L1-2 (11.0 g, 2.1 mmol) in EtOAc (90 mL) was added 10% Pd/C (0.5 g). The resulting mixture was stirred under a hydrogen balloon for one day. The mixture was then filtered through Celite. The Celite was rinsed with EtOAc (25 mL x 3). The combined filtrate was evaporated to give ethyl 3-pentyloctanoate L1-3 as a light -yellow oil (9.0 g, 83%). 1 H-NMR (300 MHz, CDCl3) δ 4.10 (q, J = 7.1 Hz, 2H), 2.20 (d, J = 6.8 Hz, 2H), 1.82 (s, 1H), 1.40-1.12 (m, 19H), 0.88 (t, J = 7.0 Hz, 6H). [1438] Synthesis of 3-pentyloctan-1-ol (L1-4) [1439] To a 2.0 M THF solution of lithium aluminum hydride (28 mL, 56 mmol) was slowly added a solution of L1-3 (7.0 g, 29 mmol) in THF (33 mL) at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 0 °C for 1h then at room temperature overnight. With ice- water bath cooling, the reaction was quenched by adding saturated aqueous Na2SO4 solution to give a milky solution. The organic phase was separated, and the aqueous phase was extracted with Et2O (50 mL x 2). The combined organic phases were dried over Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO2: 0 to 15% ethyl acetate in hexane gradient) to yield 3-pentyloctan-1-ol L1-4 as slightly yellow oil (4.0 g, 70%). 1 H-NMR (300 MHz, CDCl3) 3.65 (t, J =4.4 Hz, 2H), 1.51 (dd, J = 13.7 Hz, 6.8 Hz, 2H), 1.46-1.12 (m, 17H), 0.88 (t, J = 7.1 Hz, 6H). Synthesis of 4,4-bis(3,7-dimethyloctyl)oxy)butane nitrile (L4L-2) [Procedure A] [1440] To a 100 mL round bottom flask, 4,4-dimethoxybutanenitrile (3.0 g, 23.2 mmol), alcohol (11.0 g, 69.7 mmol) and pyridinium p-toluenesulfonate (0.29 g 1.2 mmol) were added. The resulting mixture was stirred at 120 °C for 4h and cooled to room temperature. EtOAc (50 mL) and H 2 O (20 mL) were added in, and the resulting phases were separated. The aqueous phase was extracted with EtOAc (50 mL). Combined organic extracts were washed with H2O (20 mL) and dried over anhydrous MgSO 4 . Filtration and concentration provided crude material which was purified by flash column chromatography (SiO2: 0 to 10% ethyl acetate in hexanes gradient) to yield L4L-2 as colorless oil (6.6 g, 74%); 1 HNMR (CDCl 3 ) δ 4.50-4.53 (t, 1H), 3.58-3.60 (m, 2H), 3.41 – 3.49 (m, 2H), 2.39 – 2.44 (t, 2H), 1.92-1.94 (q, 2H), 1.50- 1.55 (m, 6H), 1.38-1.42 (m, 2H), 1.11 – 1.14 (m, 14H) 0.88-0.84 (t, 18H); CIMS m/z [M+H] + 381. Synthesis of 4,4-bis((3,7-dimethyloctyl) oxy) butanoic acid (L4L-3) [Procedure B] [1441] To a 100 mL round bottom flask containing a solution of L4L-2 (8.2 g, 21 mmol) in ethanol (50 mL) was added a solution of KOH (3.6 g, 64 mmol) in water (50 mL). After completion of addition, the mixture was stirred at 120 °C for 20h. The volatiles were removed, and the reaction pH was adjusted to 5. EtOAc (150 mL) and H2O (60 mL) were added, and the resulting phases were separated. The aqueous phase was extracted with EtOAc (50 mL). Combined organic extracts were washed with H2O (60 mL x 2) and dried over anhydrous MgSO 4 . Filtration and concentration provided L4L-3 (6.4 g, 74%) which was used for the next step without further purification. 1 HNMR (CDCl3) δ 4.54 (t, 1H), 3.60-3.65 (m, 2H), 3.45- 3.49 (m, 2H), 2.39 – 2.44 (t, 2H), 1.92 – 1.94 (m, 2H), 1.50 – 1.95 (m, 6H), 1.26 – 1.55 (m, 8H), 1.11 – 1.14 (m, 6H).0.84 – 0.88 (d, 18H); CIMS m/z [M-H]- 399.
Synthesis of Select Compounds Example 1. Synthesis of (1-(4-hydroxybutyl) pyrrolidine-3,4-diyl) bis(butane-4,1-diyl) bis(2-hexyldecanoate) (Compound CY43) Synthesis of 2,3,3a,4,7,7a-hexahydro-1H-isoindole (L19-2) [1442] To a stirred solution of 3a,4,7,7a-tetrahydro-1H-isoindole-1,3(2H)-dione L19-1 (10 g, 66.1 mmol) in THF (200 mL) cooled to 0° C, 2 M lithium aluminum hydride in THF (82.5 mL, 165 mmol) was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred for 12 h. After consumption of starting materials as observed by TLC, the reaction mixture was cooled to 0° C and quenched with THF/water (40 mL, v/v 9:1) followed by 15% aq. solution of NaOH (40 mL) and water (100 mL) over 2h. The resulting mixture was stirred at room temperature for 1 h and filtered through Celite followed by washing with DCM (3x100 mL). The collected filtrate was concentrated under reduced pressure to afford L19-2 (5.8 g, 71%) as brown liquid which was used for next step without further purification. CIMS m/z 124.2 [M+H] + . Synthesis of tert-butyl 1,3,3a,4,7,7a-hexahydro-2H-isoindole-2-carboxylate (L19-3) [1443] A solution of crude L19-2 (5.8 g, 47.1 mmol) in THF (100 mL) was cooled to 0 o C under nitrogen. Triethylamine (9.8 mL, 70.6 mmol) and di-tert-butyl decarbonate (11.4 g, 52.2 mmol) were added, the reaction mixture was stirred at room temperature for 12 h. Water and DCM was added, and the aqueous phase was extracted with DCM. The organic extract was washed with saturated aqueous sodium bicarbonate and dried with Na 2 SO 4 . Filtration and concentration provided L19-3 as colorless oil (7.5 g, 71%). 1 H-NMR (300 MHz, CDCl3) δ 5.62 (s, 2H), 3.42-3.33 (m, 2H), 3.17-3.03 (m, 2H), 2.30-2.16 (m, 4H), 1.91-1.85 (m, 2H), 1.44 (s, 9H). Synthesis of tert-butyl 3,4-bis(2-oxoethyl) pyrrolidine-1-carboxylate (L19-4) [1444] L19-3 (3.0 g, 13.4 mmol, 1 eq) was dissolved in DCM (200 mL), and the solution was cooled to -78°C. Ozone was bubbled in until the color of the solution turned to blue. The reaction was then quenched with Dimethyl sulfide and stirred under nitrogen for 30 min. Removal of solvent under reduced pressure gave a crude material which was used for next step without further purification (2.31 g, 67%). Synthesis of diethyl 4,4'-(1-(tert-butoxycarbonyl) pyrrolidine-3,4-diyl) (2E,2'E)-bis(but-2- enoate) (L19-5) [1445] To a solution of triethyl phosphonoacetate (11.2 g, 50.1 mmol) in THF (60 mL) cooled to -15°C under nitrogen, was added dropwise of 1 M NaHMDS (10.1 mL, 50.1 mmol). After completion of addition, the mixture was stirred at the same temperature for 30 min then at 0°C for 60 min. The resulted mixture was slowly added to crude L19-4 (3.2 g, 12.5 mmol) at 0°C. The reaction mixture was allowed to room temperature and stirred overnight. The reaction was quenched with aqueous ammonium chloride and extracted with ethyl acetate and dried over anhydrous Na 2 SO 4 . Filtration and concentration provided crude material which was purified by flash column chromatography (SiO2: 0 to 35% ethyl acetate in hexane gradient) to yield L19-5 as colorless oil (1.01 g, 20%). 1 H-NMR (300 MHz, CDCl3) δ 6.91-6.82 (m, 2H), 5.88- 5.83 (m, 2H), 4.22-4.14 (m, 4H), 3.43-3.37 (m, 2H), 3.18-3.09 (m, 2H), 2.36-2.11 (m, 6H), 1.44 (s, 9H), 1.28 (t, 6H); CIMS m/z 296.1 [M-Boc+H] + . Synthesis of diethyl 4,4'-(1-(tert-butoxycarbonyl) pyrrolidine-3,4-diyl) dibutyrate (L19-6) [1446] To a solution of compound L19-5 (0.58 g, 1.46 mmol) in ethyl acetate (20 mL), 10% P/C (0.2 g) was added. The mixture was stirred at room temperature under hydrogen balloon for 12 h and was filtered through a pad of Celite. After washed with ethyl acetate, the filtrates were concentrated, and crude was used for next step without further purification (0.57 g, 97%). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.15-4.08 (m, 4H), 3.40-3.30 (m, 2H), 3.15-3.01 (m, 4H), 2.29 (t, 4H), 2.09-2.03 (m, 2H), 1.65-1.52 (m, 6H), 1.44 (s, 9H), 1.24 (t, 6H); CIMS m/z 300.2 [M- Boc+H] + . Synthesis of diethyl 4,4'-(pyrrolidine-3,4-diyl) dibutyrate TFA salt (L19-7) [1447] To a solution of compound L19-6 (0.57 g, 1.42 mmol) in DCM (5 mL) was added TFA (5 mL) and the mixture was stirred at room temperature for 12 h. Volatile components were removed under reduced pressure and the crude product was used for next step without further purification (0.57 g, TFA salt). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.15-4.08 (m, 4H), 3.37-3.10 (m, 4H), 2.35-2.30 (m, 7H), 1.61-1.43 (m, 7H), 1.24 (t, 6H); CIMS m/z 300.2 [M-Boc+H] + . Synthesis of diethyl 4,4'-(1-(4-(benzyloxy) butyl) pyrrolidine-3,4-diyl) dibutyrate (L19-8) [1448] To a solution of compound L19-7 (460 mg, 1.5 mmol) and benzyl 4-bromobutyl ether (411 mg, 1.69 mmol) in CPME (5 mL) and ACN (5 mL) under nitrogen was added K2CO3 (850 mg, 6.1 mmol) and KI (255 mg, 1.53 mmol). The reaction mixture was heated at 60 o C for 18 h. After cooled to room temperature, the reaction mixture was filtered through Celite, washed with ethyl acetate, and the solvent removed under vacuum to give the crude product which was purified by flash chromatography. (40 g SiO2: 0 to 10% methanol in dichloromethane gradient) to obtain compound L19-8 as colorless oil (0.41 g, 57%). 1 H-NMR (300 MHz, CDCl3) δ 7.30-7.25 (m, 5H), 4.43 (s, 2H), 4.09-4.04 (m, 4H), 3.60-3.46 (m, 4H), 3.13-3.06 (m, 4H), 2.29 (t, 4H), 1.75-1.33 (m, 14H), 1.22 (t, 6H); CIMS m/z 462.2 [M+H] + . Synthesis of 4,4'-(1-(4-(benzyloxy) butyl) pyrrolidine-3,4-diyl) bis(butan-1-ol) (L19-9) [1449] To a solution of compound L19-8 (0.4 g, 0.88 mmol) in THF (10 mL) cooled to 0° C was added dropwise 1M lithium aluminum hydride in THF (1.1 mL, 1.1 mmol). The reaction mixture was allowed to room temperature and stirred for 12 h. After consumption of starting materials as observed by TLC, the reaction mixture was cooled to 0° C and diluted with THF and quenched with 15% NaOH solution. The resulting mixture was stirred at room temperature for 1h and filtered through Celite, followed by washing with ethyl acetate. The filtrates were concentrated to give crude product (0.21 g, 62%) which was used for next step without further purification. CIMS m/z 378.3 [M+H] + . Synthesis of (1-(4-(benzyloxy) butyl) pyrrolidine-3,4-diyl) bis(butane-4,1-diyl) bis(2- hexyldecanoate) (L19-10) [1450] To a solution of compound L19-9 (200 mg,0.53 mmol) in dichloromethane (6 mL) was added DMAP (65 mg, 0.53 mmol) and EDC (0.609 g, 3.18 mmol), followed by the addition of acid L12-1 (0.135 g, 0.53 mmol). The reaction mixture was stirred at room temperature for 24h and evaporated under vacuum. The residue was dissolved in dichloromethane (100 mL) and washed with brine (80 mL x 3). After dried over anhydrous Na2SO4, the solvent was evaporated, and the crude was purified by column chromatography (40 g SiO 2 : 0 to 10% methanol in dichloromethane gradient) to obtain compound L19-10 as colorless oil. (0.41 g, 57%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.33-7.26 (m, 5H), 4.46 (s, 2H), 4.04 (t, 4H), 3.48 (t, 2H), 2.91 (s, 2H), 2.33-2.24 (m, 4H), 1.88-1.23 (m, 68H), 0.85 (t, 12H); CIMS m/z 854.7 [M+H] + . Synthesis of (1-(4-hydroxybutyl) pyrrolidine-3,4-diyl) bis(butane-4,1-diyl) bis(2- hexyldecanoate) (Compound CY43) [1451] To a solution of compound L19-10 (125 mg, 0.14 mmol) in ethyl acetate (3 mL), was added 10% P(OH) 2 /C (50 mg). The reaction mixture was stirred under hydrogen balloon at room temperature for 6 h. The mixture was filtered through a pad of Celite, the filtrates were concentrated, and the crude was purified by column chromatography (12 g SiO 2 : 0 to 10% methanol in dichloromethane gradient) to obtain compound CY43 as colorless oil (43 mg, 38%). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.05 (t, 4H), 3.64 (t, 2H), 3.30 (s, 1H), 2.91 (s, 2H), 2.33- 2.24 (m, 4H), 1.87-1.23 (m, 68H), 0.85 (t, 12H); CIMS m/z 864.7 [M+H] + . Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 o C, detector: ELSD, tR = 11.7 min, purity: 97.66%; UPLC column: Thermo Scientific Hypersil GOLD C4, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2 o C, detector: CAD, tR = 14.0 min, purity: 88.54%.
Example 2. Synthesis of (1-(4-Hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2-hexyldecanoate) (Compound CY61) Synthesis of 9-benzyl-2,4-dioxo-3,9-diazaspiro[5.5]undecane-1,5-dicarboni trile (L20-2') [1452] To ice-cooled 7M ammonia in methanol (120 mL) was added 1-benzyl-4-piperidone (40 g, 212 mmol) followed by ethyl cyanoacetate (45 mL, 2 mmol). The resulted mixture was allowed to stand in refrigerator at -2˚ C for five days. The precipitates were filtered and washed with cold methanol. Oven drying overnight provided L20-2' as off-white solid (23 g, 30%); CIMS m/z [M+H] + 323. Synthesis of diethyl 2,2'-(1-benzylpiperidine-4,4-diyl)diacetate (L20-2) [1453] A mixture of L20-2' (5.0 g, 1.6 mmol), water (5.1 mL) and conc. sulfuric acid (6 mL) was heated at 100 °C for 48 hours. After cooled to room temperature, ethanol (60 mL) was added to the mixture and it was concentrated. The procedure was repeated four times. Ethanol (40 mL) was then added to the crude product and the solution was heated under reflux for 3 days. After ice-cooling, Na 2 CO 3 (6 g) and water were added, and the mixture was concentrated. Ethyl acetate was added and the solution was washed with water and brine and dried over anhydrous Na 2 SO 4 . Filtration and concentration provided crude material which was purified by flash chromatography (SiO2: ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield L20-2 as light-yellow oil (1.85 g, 82%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.32- 7.21 (m, 5H), 4.11 (q, J = 6.5 Hz, 4H), 3.50 (s, 2H), 2.56 (s, 4H), 2.43 (t, J = 5.5 Hz, 4H), 1.68 (t, J = 6.8 Hz, 4H), 1.24 (t, J = 7.1 Hz, 6H); CIMS m/z [M+H] + 348. Synthesis of 2,2'-(1-benzylpiperidine-4,4-diyl)bis(ethan-1-ol) (L22-1) [1454] To an ice-cooled solution of 2.0 M lithium aluminum hydride in THF (5.0 mL, 10 mmol) was added slowly a solution of L20-2 (1.85 g, 5.3 mmol) in anhydrous THF (25 mL) under nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight. With ice-water bath cooling, water (0.38 mL), 15% aqueous sodium hydroxide solution (0.38 mL) and water (1.15 mL) were added successively. Filtration through Celite and concentration to yield L22-1 as an oil which slowly solidified to an off-white solid (1.32 g, 94%). 1 H-NMR (300 MHz, CDCl3) δ 7.39-7.18 (m, 5H), 3.74 (t, J = 6.5 Hz, 4H), 3.49 (s, 2H), 2.40 (t, J = 5.2 Hz, 4H), 1.67 (t, J = 6.8 Hz, 4H), 1.50 (t, J = 7.1 Hz, 6H); CIMS m/z [MH + ] 264. Synthesis of (1-benzylpiperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2-hexyldecanoate) (L22-2) [1455] To a solution of L22-1 (1.32 g, 5 mmol) in DCM (50 mL) was added L12-1 (3.4 g, 13 mmol) followed by DMAP (0.61 g, 5 mmol) and EDC (3.7 g, 20 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 48h. The reaction mixture was diluted with DCM (50 mL) and washed with saturated NaHCO3 aqueous solution (50 mL), water (25 mL) and brine (25 mL). The organic phase was dried over anhydrous Na 2 SO 4 . Filtration and concentration provided crude material which was purified by flash column chromatography (SiO 2 : ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield L22-2 as an oil which slowly solidified to a white solid (2.6 g, 70%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.31-7.19 (m, 5H), 4.12 (q, J = 7.1 Hz, 4H), 3.49 (s, 2H), 2.49-2.22 (m, 6H), 1.73-1.12 (m, 56H), 0.87 (t, J = 6.3 Hz, 12H); CIMS m/z [M+H] + 740. Synthesis of piperidine-4,4-diylbis(ethane-2,1-diyl) bis(2-hexyldecanoate) (L22-3) [1456] To a solution of L22-2 (2.6 g, 3.5 mmol) in 2-propanol (60 mL) was added 10% Pd/C (1.5 g) and 1M HCl in EtOAc (10 mL). The resulting mixture was stirred under a hydrogen balloon and heated in oil bath at 80 ˚C for 20h. The reaction mixture was filtered through Celite. The Celite was rinsed with 2-propanol, dichloromethane and EtOAc. The combined filtrate was evaporated to give L22-3 as a light -yellow oil (2.2 g, 95%); 4.20-4.00 (m, 4H), 3.49-2.90 (m, 4H), 2.35-2.15 (m, 2H), 1.95-0.90 (m, 56H), 0.87 (t, J = 6.6 Hz, 12H); CIMS m/z [M+H] + 650. Synthesis of (1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)bis(ethane-2,1-di yl) bis(2- hexyldecanoate) (L22-4) [1457] To a solution of L22-3 (1.5 g, 2.3 mmol) and 4-benzyloxybutanal (0.8 g, 4.6 mmol) in dichloroethane (60 mL) was added sodium triacetoxyborohydride (1.5 g, 6.9 mmol) followed by acetic acid (0.16 mL, 2.3 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for two days. The reaction mixture was diluted with DCM (40 mL) and washed with saturated NaHCO 3 aqueous solution (50 mL), water (25 mL) and brine (25 mL). The organic phase was dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO 2 : 0 to 100% ethyl acetate in hexane gradient) to yield L22-4 as slightly yellow oil (0.9 g, 48%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.35-7.21 (m, 5H), 4.49 (s, 2H), 4.13 (q, J = 7.1 Hz, 4H), 3.47 (t, J = 5.7 Hz, 2H), 2.49-2.20 (m, 8H), 1.75-1.12 (m, 60H), 0.87 (t, J = 6.0 Hz, 12H); CIMS m/z [M+H] + 812. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2- hexyldecanoate) (Compound CY61) [1458] To a solution of L22-4 (0.9 g, 2.1 mmol) in EtOAc (40 mL) was added 10% Pd/C (0.5 g) and 1M HCl in EtOAc (8 mL). The resulting mixture was stirred under a hydrogen balloon overnight. It was then filtered through Celite. The Celite was rinsed with EtOAc (25 mL x 3). Concentration provided crude material which was purified by flash column chromatography (SiO2: ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield Compound CY61 as a light -yellow oil (130 mg, 16%). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.12 (q, J = 7.1 Hz, 4H), 3.55 (m, 2H), 2.55-2.20 (m, 8H), 1.75-1.12 (m, 60H), 0.87 (t, J = 6.3 Hz, 12H); MS (CI): m/z [M+H] + 722.6; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 o C, detector: ELSD, tR = 11.2 min, purity: > 99%; UPLC column: Thermo Scientific Hypersil GOLD C4, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2°C, detector: CAD, t R = 12.1 min, purity: 99.21%. The acetylated product CY62 (550 mg) was also isolated as slightly yellow oil. 1 H NMR (300 MHz, CDCl 3 ): δ ppm 4.13 (q, J = 7.1 Hz, 4H), 4.06 (q, J = 6.3 Hz, 2H), 2.46-2.21 (m, 8H), 2.03 (s, 3H), 1.72-1.15 (m, 60H), 0.87 (t, J = 6.3 Hz, 12H); MS (CI): m/z [M+H] + 764.6; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2°C, detector: ELSD, tR = 11.3 min, purity: 99.83%; UPLC column: Thermo Scientific Hypersil GOLD C4, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.7 min, purity: 97.57%.
Example 3. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2-hexyldecanoate) (Compound CY57) [1459] Synthesis of tert-butyl 3,5-bis(4-(benzyloxy)but-1-en-1-yl)piperidine-1-carboxylate (L21-3) [1460] To a dry ice-acetone bath cooled solution of L21-1 (500 mg, 1.6 mmol) in anhydrous toluene (8 mL) was added 1.0 M diisobutylaluminum hydride in toluene (3.4 mL, 3.4 mmol) under nitrogen atmosphere. The resulted mixture was stirred at -72°C for 2h. About half of a pre-cooled (-72°C) solution of benzyloxypropylidene triphenylphosphorane (“Wittig Reagent”, obtained by adding potassium tert-butoxide (1.1 g, 9.3 mmol) to a solution of (3- benzyloxypropyl)triphenyl phosphonium bromide L21-2 (4.86 g, 9.6 mmol) in anhydrous toluene (8 mL) at 0°C) was stirred at room temperature for 2h. The reaction mixture was warmed to room temperature and stirred for 16h. The rest of the solution of Wittig reagent was added, and the reaction was stirred at room temperature for another 16h. The reaction was then quenched by adding water (15 mL) and extracted with ethyl acetate (25 mL x 3). Combined organic extracts were washed with water (25 mL x 3) and dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO2: ethyl acetate/hexane 0-100) to yield L21-3 as colorless oil (250 mg, 30%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.39-7.18 (m, 10H), 5.55-5.40 (m, 2H), 5.17 (t, J = 9.1 Hz, 2H), 4.51 (s, 4H), 3.99 (s, br, 4H), 3.49 (t, J = 6.9 Hz, 4H), 2.58-2.23 (m, 6H), 1.77-1.68 (m, 1H), 1.45 (s, 9H), 1.09-0.96 (m, 1H); CIMS m/z [M-Boc+H] + 405.7. Synthesis of tert-butyl 3,5-bis(4-hydroxybutyl)piperidine-1-carboxylate (L21-4) [1461] A mixture of L21-3 (470 mg, 0.9 mmol) and 10% Pd/C (100 mg) in methanol (12 mL) was stirred under a hydrogen balloon at room temperature for 20h. The reaction mixture was filtered through Celite. The Celite was washed with methanol. The combined filtrate was evaporated to give L21-4 as a light yellow oil (300 mg, 98%); 4.20-3.95 (m, 4H), 3.63 (t, J = 6.3 Hz, 4H), 2.25-2.05 (m, 2H), 1.93-1.82 (m, 1H), 1.70-1.05 (m, 23H), 0.69-0.53 (m, 1H); CIMS m/z [M-Boc+H] + 230. Synthesis of (1-(tert-butoxycarbonyl)piperidine-3,5-diyl)bis(butane-4,1-d iyl) bis(2- hexyldecanoate) (L21-5) [1462] To a solution of L21-4 (300 mg, 0.9 mmol) in DCM (10 mL) was added L12-1 (580 mg, 2.3 mmol) followed by DMAP (110 mg, 0.9 mmol) and EDC (700 mg, 3.6 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 20h. The reaction mixture was diluted with DCM (15 mL) and washed with brine (10 mL). The organic phase was dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO 2 : ethyl acetate/hexane 0-100%) to yield L21-5 as colorless oil (600 mg, 82%). 1 H-NMR (300 MHz, CDCl3) δ 4.20-3.95 (m, 4H), 4.05 (t, J = 6.6 Hz, 4H), 2.39-2.22 (m, 2H), 2.21-2.05 (m, 2H), 1.91-1.80 (m, 1H), 1.68-1.11 (m, 69H), 0.86 (t, J = 6.3 Hz, 12H) 0.69-0.53 (m, 1H); CIMS m/z [M-Boc+H] + 706.7. Synthesis of piperidine-3,5-diylbis(butane-4,1-diyl) bis(2-hexyldecanoate) (L21-6) [1463] To a solution of L21-5 (450 mg, 0.56 mmol) in dichloromethane (3 mL) was added TFA (3 mL) at 0°C and the reaction mixture was stirred at room temperature for 4 h. The volatile components were removed under reduced pressure and the crude L21-6 (450 mg) was used for the next step without further purification. 1 H-NMR (300 MHz, CDCl3) δ 4.05 (t, J = 6.3 Hz, 4H), 3.49–2.80 (m, 4H), 2.51–2.22 (m, 4H), 2.02–1.01 (m, 61H), 0.69-0.53 (m, 13H); CIMS m/z [M+H] + 706.7. Synthesis of (1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)bis(ethane-2,1-di yl) bis(2- hexyldecanoate) (L21-7) [1464] To a solution of L21-6 (450 mg, 0.55 mmol) and 4-benzyloxybutanal (198 mg, 1.1 mmol) in 1,2-dichloroethane (15 mL) was added sodium triacetoxyborohydride (354 mg, 1.6 mmol) followed by acetic acid (36 µL, 0.55 mmol). The resulted mixture was stirred at room temperature under nitrogen atmosphere for 20h. The reaction mixture was diluted with DCM (20 mL) and sat. aq. sodium bicarbonate solution was slowly added until no bubbles produced. The resulted two phases were separated and the aqueous phase was extracted with DCM (20 x 2 mL). Combined organic extracts were dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO2: ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield L21-7 as slightly yellow oil (340 mg, 71%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.35-7.21 (m, 5H), 4.49 (s, 2H), 4.05 (t, J = 6.3 Hz, 4H), 3.47 (m, 2H), 2.95-2.92 (m, 2H), 2.50-2.10 (m, 6H), 1.81-1.70 (m, 1H), 1.65-1.15 (m, 66H), 0.86 (t, J = 6.9 Hz, 12H), 0.55-0.42 (m, 1H); CIMS m/z [M+H] + 868. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2- hexyldecanoate) (Compound CY57) [1465] A mixture of L21-7 (340 mg, 0.4 mmol) and 10% Pd(OH)2/C (120 mg) in EtOAc (12 mL) was stirred under a hydrogen balloon for 70h. It was then filtered through Celite. The Celite was rinsed with EtOAc (10 mL x 3). Concentration provided crude material which was purified by flash column chromatography (SiO2: ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield Compound CY57 as a light -yellow oil (171 mg, 56%). 1 H-NMR (300 MHz, CDCl3) δ 4.04 (t, J = 6.3 Hz, 4H), 3.54 (m, 2H), 2.95-2.92 (m, 2H), 2.61- 2.22 (m, 6H), 1.85-1.75 (m, 1H), 1.76-1.12 (m, 66H), 0.87 (t, J = 6.9 Hz, 12H), 0.55-0.42 (m, 1H); MS (CI): m/z [M+H] + 778.7; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2°C, detector: ELSD, t R = 11.6 min, purity: > 99%; UPLC column: Thermo Scientific Hypersil GOLD C4, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2°C, detector: CAD, t R = 13.3 min, purity: 97.05%. Example 4. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(octyloxy) butanoate) (CY63) Synthesis of diethyl 2,2'-(piperidine-4,4-diyl)diacetate (L27-1) A solution of L20-2 (5.4 g, 15.4 mmol) in ethanol (107 ml) at room temperature was treated with 10% Pd/C (1.1 g) under nitrogen atmosphere. The reaction mixture was evacuated and flushed with H2 gas (3x) and then stirred vigorously under an atmosphere of H2 (1 atm, H2 - balloon) at room temperature. After 24 h, the reaction mixture was filtered through Celite and the filtrate was concentrated in vacuo to give the crude product, L27-1 (4 g) which was used for the next step without further purification. APCI MS m/z [M+H] + 257.16. Synthesis of diethyl 2,2'-(1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)diacetate (L27-2 To a mixture of L27-1 (4 g, 15.5 mmol) and 4-(benzyloxy)butanal (5.5 g, 31.1 mmol) in 1,2- dichloroethane (180 mL) was added Na(OAc) 3 BH (9.9 g, 46.6 mmol) and acetic acid (1 mL). The reaction mixture was subjected to vacuum/N2 cycle (3x) and stirred at room temperature for 18 h. The reaction was quenched by slow addition of saturated NaHCO 3 (100 mL) at 0 °C. The aqueous phase was extracted using ethyl acetate (100 mL, 3x) and the combined organic phases were dried over anhydrous Na 2 SO 4 . Filtration followed by concentration provided crude material, which was dissolved in DCM. Silica gel (40 g) and triethyl amine (40 mL) were added to the crude material and shaken for 10-15 min and the solvent was removed under vacuum. The residue was loaded on to an empty flash cartridge, which was then attached to flash purification system loaded with 80 g flash silica column and was purified by flash chromatography (SiO 2 : 0 to 10% ethyl acetate in hexane (10% triethylamine)) to yield ethyl L27-2 as slightly yellow oil (3.7 g, 57%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.31-7.30 (m, 5H), 4.46 (s, 2H), 4.09-4.04 (m, 4H), 3.47-3.43 (m, 2H), 2.52-2.31 (m, 10H), 1.68-1.57(m, 8H), 1.22 (t, 6H); APCI MS m/z [M+H] + 420.3. Synthesis of 2,2'-(1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)bis(ethan-1- ol) (L27-3) A solution of L27-2 (0.75 g, 1.78 mmol) in THF (14 mL) was cooled in an ice bath (0 °C) and to this was added 2M LiAlH 4 in THF (3.56 mL, 7.14 mmol), dropwise. The ice bath was removed, and the reaction mixture was stirred for 18 h at room temperature. The mixture was diluted with Et 2 O (50 mL), cooled in an ice bath, and carefully quenched with water (10 mL), 20% NaOH (10 mL) and water (30 mL). After stirring for 30 min, the aqueous phase was extracted with 20 mL DCM (3x), then the combined organic phase was dried (Na 2 SO 4 ), filtered and concentrated to give L27-3 (0.54 g, 91% yield) as a white solid. APCI MS m/z [M+H] + 336.3. Synthesis of 4,4-bis(nonyloxy)butanoic acid (L4-3(T9)) [1466] Prepared following Procedure B described in Compound L4L synthesis. Compound L4-3(T9) was isolated as light-yellow oil in a yield of 11.8 g (98%). 1 HNMR (CDCl3) δ: 4.53- 4.56 (t, 1H), 3.57–3.60 (m, 2H), 3.40–3.43 (m, 2H), 2.39–2.41 (t, 2H), 1.90–1.95 (m, 2H), 1.54–1.56 (M, 4H), 1.26 (bs, 28H), 0.85–0.87 (t, 6H); CIMS m/z [M-H]- 371. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(octyloxy) butanoate) (L27-4) [Procedure E] [1467] To a 250mL round bottom flask containing L4-3(T9) (1 g, 2.9 mmol, 2.5 eq), EDC (1.01 g, 5.28 mmol, 4 eq), DMAP (161 mg, 1.32 mmol, 1 eq) and L27-3 (440 mg, 1.32 mmol, 1 eq) was added anhydrous dichloromethane (20 mL) and the reaction mixture was stirred at room temperature overnight. After completion of the reaction about 30g of flash silica was added and the contents were stirred well to get a uniform mixture. Solvent was removed from this mixture under vacuum. The residue was loaded on to an empty flash cartridge, which was then attached to a flash purification system loaded with flash silica column and was purified by flash chromatography (SiO2: hexane (10% triethyl amine)/ethyl acetate 0-20%) to get Compound L27-4 (0.94 g, 73%) as slightly yellow oil. 1 H-NMR (300 MHz, CDCl 3 ) δ 7.33- 7.31 (m, 5H), 4.48-4.47 (m, 4H), 4.10-4.08 (m, 4H), 3.56-3.37 (m, 10H), 2.37-2.32 (m, 10H), 1.90-1.80(m, 20H), 1.31-1.10(m, 40H), 0.84 (t, 12H); APCI MS m/z [M+H] + 988.8. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(octyloxy) butanoate) (CY63) [Procedure F] [1468] To a 250 mL round bottom flask containing L27-4 (560 mg, 0.56 mmol) and 10% Pd/C (186 mg) was added ethyl acetate (20 mL) and then the reaction mixture was subjected to vacuum/N 2 cycle (3x) followed by another cycle of vacuum/H 2 (3x). The reaction mixture was placed under 1 atm H2 (hydrogen balloon) and left to stir overnight. The reaction mixture was diluted with ethyl acetate (100 mL) and then filtered through Celite, washed with ethyl acetate, and then the solvent was removed under vacuum to dryness to give the crude product as a light brown oil CY63 (132 mg, 26%). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.48 (t, 4H), 4.12-4.07 (m, 4H), 3.55-3.39 (m, 10H), 2.46-2.31 (m, 10H), 1.90-1.88(m, 4H), 1.66-1.51(m, 20H), 1.30- 1.00(m, 40H), 0.86 (t, 12H); APCI MS m/z [M+H] + 898.8; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: ELSD, tR = 11.6 min, purity: >99 %; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.8 min, purity: > 99%.
Example 5. Synthesis of heptadecan-9-yl 2-(1-(4-hydroxybutyl)-4-(2-oxo-2-((3- pentyloctyl)oxy)ethyl) piperidin-4-yl)acetate (Compound CY69) Synthesis of 2,2'-(1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)diacetic acid (L28A-1) [1469] To a solution of diester L27-2 (0.9 g, 2.1 mmol) in THF (15 mL) and methanol (2.5 mL) was added a solution of LiOH (0.36 g, 6.4 mmol) in water (5 mL). The mixture was stirred at room temperature for 20 h. While cooling in ice-water bath, the reaction mixture pH was adjusted to 4. Volatile components were removed under reduced pressure and the residue was lyophilized to give an off-white solid which was purified by reverse column chromatography (acetonitrile/water 0-100) to yield L28A-1 as off-white foam solid (0.62 g, 80%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.39-7.18 (m, 5H), 4.47 (s, 2H), 3.48 (t, J = 6.3 Hz, 2H), 2.90-2.56 (m, 6H), 2.43 (m, 4H), 2.05-1.56 (m, 8H); CIMS m/z [M-H]- 361.5. Synthesis of 9-(4-(benzyloxy)butyl)-3-oxa-9-azaspiro[5.5]undecane-2,4-dio ne (L28A-2) [1470] To a solution of L28A-1 (0.5 g, 1.4 mmol) in anhydrous DCM (15 mL) and pyridine (2 mL) at 0 °C under nitrogen atmosphere was added anhydrous DMF (1 drop) and oxalyl chloride (0.15 mL, 4.2 mmol). After completion of the addition, the mixture was stirred at room temperature for 18 h. More oxalyl chloride (0.15 mL, 4.2 mmol) was added and the mixture was stirred at room temperature for 20 h. The reaction mixture was concentrated and co- evaporated with anhydrous toluene to give L28A-2 as a light yellow oil (0.48 g, 99%); CIMS m/z [M+H] + 346.2. Synthesis of 2-(1-(4-(benzyloxy)butyl)-4-(2-oxo-2-((3-pentyloctyl)oxy)eth yl)piperidin-4- yl)acetic acid (L28A-3) [1471] To a solution of L28A-2 (480 mg, 1.4 mmol) in DCM (15 mL) and pyridine (2 mL) at 0 °C was added L1-4 (800 mg, 4.0 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 18h. More L1-4 (160 mg, 1.4 mmol) was added, and the mixture was stirred at 50 °C for 20 h. The reaction mixture was concentrated, and the crude material was purified by flash column chromatography (SiO 2 : Methanol/DCM 0-30% with 5% triethylamine) to yield L28A-3 as light-yellow solid (330 mg, 44%). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.41-7.15 (m, 5H), 4.46 (s, 2H), 4.06 (t, J = 6.5 Hz, 2H), 2.89-2.36 (m, 10H), 1.97-1.15 (m, 29H), 0.87 (t, J = 6.8 Hz, 6H); CIMS m/z [M+H] + 546.4. Synthesis of heptadecan-9-yl 2-(1-(4-(benzyloxy)butyl)-4-(2-oxo-2-((3-pentyloctyl)oxy)eth yl) piperidin-4-yl)acetate (L28A-4) [1472] To a solution of L28A-3 (320 mg, 0.58 mmol) in DCM (10 mL) was added heptadecan- 9-ol (L2-1) (225 mg, 0.88 mmol) followed by DMAP (38 mg, 0.3 mmol) and EDC (225 mg, 1.2 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 18 h. The reaction mixture was diluted with DCM (15 mL) and washed with brine (10 mL). The organic phase was dried over anhydrous Na2SO4. Filtration and concentration provided crude material which was purified by flash column chromatography (SiO 2 : ethyl acetate/hexane 0-100%) to yield L28A-4 as colorless oil (305 mg, 66%). 1 H-NMR (300 MHz, CDCl3) δ 7.38- 7.21 (m, 5H), 4.84 (m, 1H), 4.49 (s, 2H), 4.05 (t, J = 7.1 Hz, 2H), 3.47 (t, J = 5.9 Hz, 2H), 2.60- 2.28 (m, 10H), 1.76-1.15 (m, 55H), 0.87 (t, J = 6.0 Hz, 12H); CIMS m/z [M+H] + 784.8. Synthesis of heptadecan-9-yl 2-(1-(4-hydroxybutyl)-4-(2-oxo-2-((3-pentyloctyl)oxy)ethyl) piperidin-4-yl)acetate (Compound CY69) [1473] A mixture of L28A-4 (300 mg, 0.38 mmol) and 10% Pd(OH)2/C (150 mg) in EtOAc (15 mL) was stirred under a hydrogen balloon for 80h. The mixture was then filtered through Celite. The Celite was rinsed with EtOAc (10 mL x 3). Concentration of the filtrate provided crude material which was purified by flash column chromatography (SiO 2 : ethyl acetate/hexane 0-100% with 1% triethylamine in the eluent) to yield Compound CY69 as a light-yellow oil (241 mg, 91%). 1 H-NMR (300 MHz, CDCl3) δ 4.84 (m, 1H), 4.05 (t, J = 7.4 Hz, 2H), 3.56 (m, 2H), 2.71-2.35 (m, 10H), 1.82-1.43 (m, 15H), 1.36-1.15 (m, 40H), 0.87 (t, J = 5.2 Hz, 12H); MS (CI): m/z [M+H] + 694.6; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: ELSD, t R = 10.6 min, purity: > 99%; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.4 min, purity: > 99%. Example 6. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2-nonylundecanoate) (CY65) Synthesis of (1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)bis(ethane-2,1-di yl) bis(2- nonylundecanoate) (L49-4) [1474] Prepared following Procedure E described in Compound L27 synthesis. Compound L49-1 was isolated as colorless oil (580 mg, 43%). 1 H-NMR (300 MHz, CDCl3) δ 7.33-7.26 (m, 5H), 4.48 (s, 2H), 4.12 (t, 4H), 3.46 (t, 2H), 2.45-2.24 (m, 7H), 1.70-1.37 (m, 19H), 1.29- 1.15 (m, 58H), 0.86 (t, 12H); CIMS m/z [M+H] + 925.56. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(2- nonylundecanoate) (CY65) [1475] Prepared following Procedure F described in Compound CY63 synthesis. Compound CY65 was isolated as colorless oil (0.52 g, 97%). 1 H-NMR (300 MHz, CDCl 3 ) δ 4.11 (m, 4H), 3.56 (t, 2H), 2.55-241 (m, 4H), 2.27-2.21 (m, 2H), 1.68-1.57 (m, 16H), 1.28-1.15 (m, 54H), 0.86 (t, 12H); CIMS m/z [M+H] + 834.1. Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: ELSD, tR = 12.1 min, purity: > 99%; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: CAD, tR = 15.4 min, purity: > 98.1%. Example 7. Synthesis of (1-(4-hydroxybutyl)piperidine-3,5-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(nonyloxy) butanoate) (CY66) Synthesis of (1-(4-(benzyloxy)butyl)piperidine-3,5-diyl)bis(ethane-2,1-di yl) bis(4,4-bis (nonyl oxy)butanoate) (L50-1) [1476] Prepared following Procedure E described in Compound CY63 synthesis. Compound L50-1 (0.55 g, 44%) was isolated as colorless oil. 1 H-NMR (300 MHz, CDCl 3 ) δ 7.33-7.32 (m, 5H), 4.49-4.47 (m, 4H), 4.12-4.10 (m, 4H), 3.56-3.37 (m, 10H), 2.38-2.33 (m, 10H), 2.04-1.89 (m, 4H), 1.66-1.50 (m, 20H), 1.40-0.99 (m, 48H), 0.87 (t, 12H); APCI- MS: m/z [M+H] + 1045.0. Synthesis of (1-(4-hydroxybutyl)piperidine-3,5-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(nonyloxy) butanoate) (CY66) [1477] Prepared following Procedure F described in Compound CY63 synthesis. Compound CY66 (0.2 g, 44%) was isolated as colorless oil. 1 H-NMR (300 MHz, CDCl 3 ) δ 4.47 (t, 4H), 4.13-4.10 (m, 4H), 3.56-3.40 (m, 10H), 2.57-2.39 (m, 10H), 1.91-1.89 (m, 4H), 1.67-1.52 (m, 26H), 1.37-1.00 (m, 48H), 0.87 (t, 12H); APCI-MS: m/z [M+H] + 954.7; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: = 11.6 min, purity: >99 %; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.8 min, purity: >99%. Example 8. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(4,4-bis(decyloxy)butanoate) (CY67) Synthesis of (1-(4-(benzyloxy)butyl)piperidine-4,4-diyl)bis(ethane-2,1-di yl) bis(4,4- bis(decyloxy)butanoate) (L51-1) [1478] Prepared following Procedure E described in Compound CY63 synthesis. L51-1 (1.16 g, 88%), colorless oil, 1 H-NMR (300 MHz, CDCl3) δ 7.33-7.25 (m, 5H), 4.51-4.46 (m, 4H), 4.11 (t, J = 7.5 Hz, 4H), 3.62-3.33 (m, 10H), 2.45-2.26 (m, 10H), 1.97-1.85 (m, 4H), 1.73-1.41 (m, 18H), 1.40-1.15 (m, 58H), 0.87 (t, J = 6.3 Hz, 12H); MS (CI): m/z [M+H] + 1100.8. Synthesis of (1-(4-hydroxybutyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl) bis(4,4- bis(decyloxy)butanoate) (CY67) [1479] Prepared following Procedure F described in Compound CY63 synthesis. Compound CY67 (615 mg, 58%), colorless oil, 1 H-NMR (300 MHz, CDCl 3 ) δ 4.48 (t, J = 5.6 Hz, 2H), 4.11 (t, J = 7.4 Hz, 4H), 3.61-3.32 (m, 10H), 2.65-2.30 (m, 10H), 1.98-1.85 (m, 4H), 1.76-1.15 (m, 76H), 0.91-0.80 (m, 12H); MS (CI): m/z [M+H] + 1010.8; Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: = 12.4 min, purity: > 99%; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 60% to 100% in 15 min, flow rate: 0.5mL/min, column temperature: 20±2 °C, detector: CAD, t R = 16.0 min, purity: 98%. Example 9. Synthesis of (1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)bis(ethane - 2,1-diyl) bis(4,4 Synthesis of 4-bromobutanal (L57-2) [1480] To a solution of pyridinium chlorochromate (PCC) (12.14 g, 56.55mmol) in DCM (75 mL) was added 4-bromobutan-1-ol (5.77 g, 37.7 mmol) in DCM (25 mL) over 10 min (intermittent cooling was required to prevent solvent reflux). The reaction mixture was stirred at room temperature for 2h and then diluted with diethyl ether. The upper ether phase was decanted from the flask and filtered through celite and the celite cake was washed with ether. Combined ether phases were evaporated under reduced pressure to get crude 4-bromobutanal L57-2, which was used for the next step without further purification (4.5 g, crude); 1 H-NMR (300 MHz, CDCl3) δ 9.81 (s, 1H), 3.74 (m, 2H), 2.18 (m, 2H), 1.84 (s, 2H). Synthesis of 4-bromo-1,1-dimethoxybutane (L57-3) [1481] 4-bromobutanal L57-2 (4.5 g, crude) was dissolved in methanol (10 mL), then 2N HCl in ether (10 mL) was then added in. The reaction mixture was stirred at room temperature overnight. The volatile components were evaporated under reduced pressure to yield 4-bromo- 1,1-dimethoxybutane L57-3 as light-yellow oil (3.9 g, crude); 1 H-NMR (300 MHz, CDCl 3 ) δ 4.38 (m, 1H), 3.4 (m, 2H), 3.32 (s, 6H), 1.91 (m, 2H), 1.75 (m, 2H). Synthesis of 1-(4,4-dimethoxybutyl)-1H-imidazole (L57-4) [1482] To a solution of imidazole (1.48 g, 21.76 mmol) in anhydrous THF (40 mL) at 5-10 °C was added NaH (948 mg, 23.74 mmol, 60% in mineral oil) portionwise with stirring. The resulting mixture was then stirred at room temperature for 2h. To the suspension was added dropwise 4-bromo-1,1-dimethoxybutane L57-3 (3.9 g, 19.79 mmol) in THF (10 mL) over a period of 15 min and the reaction was further stirred for 3 h at room temperature to achieve a uniform mixture. The reaction mixture was heated at 60 °C overnight, cooled to room temperature and filtered. THF was removed under reduced pressure and the residue was purified by flash chromatography (SiO2: 0-5% MeOH in DCM gradient) to yield 1-(4,4- dimethoxybutyl)-1H-imidazole L57-4 (850 mg, 12 % over 3 steps). 1 H-NMR (300 MHz, CDCl3) δ 7.45 (s, 1H), 7.04 (s, 1H), 6.9 (s, 1H), 4.32 (t, J = 5.49 Hz, 1H), 3.95 (t, J = 7.14 Hz, 2H), 3.29 (s, 6H), 1.84 (m, 2H), 1.58 (m, 2H); CIMS m/z [M+H] + 185. Synthesis of 1-(4-oxobutyl)-1H-imidazol-1-ium chloride (L57-5) [1483] To a solution of 1-(4,4-dimethoxybutyl)-1H-imidazole L57-4 (1.05 g, 5.7 mmol) in THF (5.0 mL), was added 1.5N HCl (5.0 mL). The reaction mixture was stirred at room temperature overnight. THF was evaporated and water layer was washed with DCM (10 mL) and EtOAc (10 mL) to remove impurities. the aqueous layer was evaporated under reduced pressure followed by co-evaporation with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) and dried under high vacuum for 24 h to yield 1-(4-oxobutyl)-1H-imidazol-1-ium chloride L57-5 as a light-yellow gummy solid which was used for the next step without further purification (1.0 g, crude). 1 H-NMR (300 MHz, DMSO-D6) δ 9.63 (s, 1H), 9.21 (s, 1H), 7.81 (s, 1H), 7.7 (s, 1H), 4.19 (m, 2H), 3.34-3.62 (m, 2H) 2.05 (m, 2H); CIMS m/z [M+H] + 139. Synthesis of diethyl 2,2'-(1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)diace tate (L57-6) [1484] To a solution of diethyl 2,2'-(piperidine-4,4-diyl)diacetate (850 mg, 3.5 mmol) in a mixture of DMF (5 mL) and DCE (5 mL) was added 1-(4-oxobutyl)-1H-imidazol-1-ium chloride L57-5 (1.0 g, 5.74 mmol) in DMF (5 mL), followed by addition of Na(OAc) 3 BH (2.22 g, 10.5 mmol) and AcOH (240 µL, 4.2 mmol). The reaction mixture was stirred at room temperature under nitrogen for 18 hours. LC-MS confirms completion of the reaction. The reaction mixture was diluted with DCM and washed with Sat. NaHCO3. Aqueous layer was extracted with DCM (3 x 50 mL). Combined organic layers were dried over Na2SO4 and concentrated under reduced pressure to get crude product which was purified by flash chromatography (SiO2: 0-6% MeOH in DCM gradient) to yield diethyl 2,2'-(1-(4-(1H- imidazol-1-yl)butyl)piperidine-4,4-diyl)diacetate L57-6 (600 mg, 45%). 1 H-NMR (300 MHz, CDCl3) δ 7.45 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.08 (q, J = 7.14 Hz, 4H), 3.93 (t, J = 7.14 Hz, 2H), 2.53 (s, 4H), 2.36 (m, 6H), 1.78 (m, 2H), 1.67 (m, 4H), 1.5 (m, 2H), 1.23 (t, J = 7.14 Hz, 6H); CIMS m/z [M+H] + 380. Synthesis of 2,2'-(1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)bis(e than-1-ol) (L57-7) [1485] To a solution of 2,2'-(1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)diace tate L57- 6 (600 mg, 1.58 mmol) in anhydrous THF (10 mL) and 0°C was added dropwise a solution of LiAlH 4 in anhydrous THF (2.0 M, 1.6 mL, 3.16 mmol) under nitrogen. The resulting reaction mixture was stirred at room temperature overnight. The reaction mixture was cooled to 0°C and Na 2 SO 4 .10H 2 O was added slowly until all gas evolution stopped. After filtration through celite, the celite cake was washed with THF. Combined filtrates were concentrated under reduced pressure to give 2,2'-(1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)bis(e than-1- ol) L57-7 as colorless viscous liquid, which was used for the next step without further purification (440 mg, crude). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.45 (s, 1H), 7.02 (s, 1H), 6.89 (s, 1H), 3.93 (t, J = 7.14 Hz, 2H), 3.7 (t, J = 6.6 Hz, 4H), 2.33 (m, 6H), 1.77 (m, 2H), 1.65 (t, J = 6.75 Hz, 4H), 1.55 (m, 2H), 1.47 (m, 4H); CIMS m/z [M+H] + 296. Synthesis of (1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4-diyl)bis(ethane -2,1-diyl) bis(4,4- bis(nonyloxy)butanoate) (CY71) [1486] To a solution of 4,4-bis(nonyloxy)butanoic acid (1.21 g, 3.27 mmol) in DCM (15 mL) was added DMAP (363 mg, 2.98 mmol) and EDC (1.25 g, 6.55 mmol). The reaction mixture was stirred at room temperature for 15 min, 2,2'-(1-(4-(1H-imidazol-1-yl)butyl)piperidine-4,4- diyl)bis(ethan-1-ol) L57-7 (440 mg, 1.49 mmol) in DCM (5 mL) was added to the reaction mixture. The reaction mixture was stirred at room temperature overnight. Formation of product was confirmed by LCMS. The reaction mixture was diluted with DCM, then washed with water and brine. The DCM layer was dried over Na2SO4 and concentrated under reduced pressure to give crude product. Crude product was purified by flash chromatography (SiO 2 : 0-5% MeOH in DCM and 1% NH4OH gradient) to yield Compound CY71 as colorless oil (404 mg, 26% after two steps). 1 H-NMR (300 MHz, CDCl 3 ) δ : 7.45 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.47 (t, J = 5.3 Hz, 2H), 4.1 (t, J = 7.4 Hz, 4H), 3.93 (t, J = 6.75 Hz, 2H), 3.53 (m, 4H), 3.4 (m, 4H), 2.35 (m, 10H), 1.89 (m, 4H), 1.75 (m, 2H), 1.66 (m, 2H), 1.54 (m, 12H), 1.25 (m, 48H), 0.87 (m, 12H); CIMS m/z [M+H] + : 1004.1. Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: ELSD, t R = 8.5 min, purity: > 99%; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.7 min, purity: > 99 %. Example 10. Synthesis of (1-(3-(1H-imidazol-1-yl)propyl)piperidine-4,4- diyl)bis(ethane-2,1-diyl) bis(4,4-bis(nonyloxy)butanoate) (CY70) Synthesis of 1-(3,3-dimethoxypropyl)-1H-imidazole (L64-2) [1487] To a solution of imidazole (2.0 g, 30 mmol) in anhydrous THF (60 mL) was added NaH (1.31 g, 32.78 mmol, 60% in mineral oil) portionwise with stirring. The resulting mixture was stirred at room temperature for 2h. To the suspension formed was added 3-bromo-1,1- dimethoxypropane (5.0 g, 27.32 mmol) in THF (15 mL) dropwise over a period of 10 min and further stirred for 3 h to achieve uniform mixture. The reaction mixture was heated at 60°C overnight and then cooled to room temperature and filtered. THF was removed under reduced pressure. DCM was added followed by addition of activated charcoal and anhydrous Na2SO4, stirred for 2 h and filtered over celite. DCM was removed under reduced pressure to get crude product as light yellowish liquid, which was purified by flash chromatography (SiO2: 0-5% in MeOH in DCM gradient) to yield 1-(3,3-dimethoxypropyl)-1H-imidazole L64-2 (3.64 g, 78%). 1 H-NMR (300 MHz, CDCl3) δ: 7.45 (s, 1H), 7.04 (s, 1H), 6.9 (s, 1H), 4.24 (t, J = 5.49 Hz, 2H), 4.01 (t, J = 7.14 Hz, 4H), 2.05 (q, J = 6.84 Hz, 4H); CIMS m/z [M+H] + 171.1. Synthesis of 3-(1H-imidazol-1-yl)propanal hydrochloride (L64--3) [1488] To a solution of compound 1-(3,3-dimethoxypropyl)-1H-imidazole L64-2 (3.0 g17.64 mmol) in THF (15.0 mL), was added 1.5N HCl (15.0 mL). The reaction mixture was stirred at room temperature overnight. THF was evaporated and water layer was washed with DCM and EtOAc to remove the impurities. The aqueous layer was evaporated under reduced pressure followed by co-evaporation with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) and dried under high vacuum for 24 h to yield 3-(1H-imidazol-1-yl)propanal hydrochloride L64-3 as light-yellow gummy solid (2.7 g) which was used for the next step without further purification. 1 H-NMR (300 MHz, DMSO-D6) δ: 9.67 (s, 1H), 9.14 (s, 1H), 7.75 (s, 1H), 7.66 (s, 1H), 4.42 (t, J = 6.45 Hz, 2H), 3.18 (t, J = 6.6 Hz, 2H); CIMS m/z [M+H] + 125.2. Synthesis of diethyl 2,2'-(1-(3-(1H-imidazol-1-yl)propyl)piperidine-4,4-diyl)diac etate (L64-4) [1489] To a solution of diethyl 2,2'-(piperidine-4,4-diyl)diacetate (555 mg, 2.28 mmol) in DCE (10 mL) was added 3-(1H-imidazol-1-yl)propanal hydrochloride L64-3 (730 mg, 4.56 mmol), followed by addition of Na(OAc) 3 BH (1.45 g, 6.84 mmol) and AcOH (156 mL, 2.73 mmol). The reaction mixture was stirred at room temperature under nitrogen for 18 hours and then was heated at 50°C and stirred for 2h. Completion of the reaction was confirmed by LCMS. The reaction mixture was diluted with DCM and washed with Sat. NaHCO3. the aqueous layer was extracted with DCM. Combined organic layers were dried over Na 2 SO 4 and concentrated under reduced pressure to get crude product. Crude product was purified by flash chromatography (SiO2: 0-6% MeOH in DCM gradient) to yield diethyl 2,2'-(1-(3-(1H-imidazol-1- yl)propyl)piperidine-4,4-diyl)diacetate L64-4 (440 mg, 52%). 1 H-NMR (300 MHz, CDCl 3 ) δ : 7.48 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.09 (q, J = 7.1 Hz, 2H), 3.99 (t, J = 6.84 Hz, 4H), 2.54 (s, 4H) 2.4 (m, 4H), 2.28 (t, J = 7.14 Hz, 2H), 1.92 (m, 2H), 1.69 (m, 4H), 1.26 (t, J = 7.14 Hz, 6H); CIMS m/z [M+H] + 366.2. Synthesis of 2,2'-(1-(3-(1H-imidazol-1-yl)propyl)piperidine-4,4-diyl)bis( ethan-1-ol) (L64-5) [1490] To a solution of diethyl 2,2'-(1-(3-(1H-imidazol-1-yl)propyl)piperidine-4,4- diyl)diacetate L64-4 (430 mg, 1.17 mmol) in anhydrous THF (10 mL) was added dropwise a solution of 2.0M LiAlH4 in THF (1.2 mL, 2.35 mmol) at 0°C. The resulting mixture was stirred at room temperature overnight. The reaction mixture was cooled to 0°C and Na 2 SO 4 .10H 2 O was added slowly until all gas evolution stopped. After filtration through celite, the celite cake was washed with THF. All filtrates were concentrated under reduced pressure to give 2,2'-(1- (3-(1H-imidazol-1-yl)propyl)piperidine-4,4-diyl)bis(ethan-1- ol) L64-5 as colorless viscous liquid (375 mg, crude), which was used for the next step without further purification. 1 H-NMR (300 MHz, CDCl3) δ 7.64 (s, 1H), 7.12 (s, 1H), 6.91 (s, 1H), 4.05 (t, J = 6.6 Hz, 2H), 3.61 (t, J = 7.6 Hz, 4H), 2.45 (m, 4H), 2.32 (t, J = 7.14 Hz, 2H), 1.9 (m, 2H), 1.56 (m, 8H); CIMS m/z [M+H] + 282.2. Synthesis of (1-(3-(1H-imidazol-1-yl)propyl)piperidine-4,4-diyl)bis(ethan e-2,1-diyl) bis(4,4- bis(nonyloxy)butanoate) (CY70) [1491] To a solution of 4,4-bis(nonyloxy)butanoic acid (L4-3(T9)) (375 mg, 1.31 mmol) in DCM (15 mL) was added DMAP (320 mg, 2.62 mmol) and EDC (1.2 g, 6.29 mmol). The reaction mixture was stirred at room temperature for 15 min, 2,2'-(1-(3-(1H-imidazol-1- yl)propyl)piperidine-4,4-diyl)bis(ethan-1-ol) L64-5 in DCM (5 mL) was added in. The reaction mixture was stirred at room temperature overnight and then diluted with DCM, washed with water and brine. DCM layer was dried over Na 2 SO 4 and concentrated under reduced pressure. Crude product was purified by flash chromatography (SiO2: 0-5% MeOH in DCM and 1% NH4OH gradient) to yield Compound CY70 as colorless oil (445 mg, 25% after two steps). 1 H-NMR (300 MHz, CDCl 3 ) δ 7.45 (s, 1H), 7.04 (s, 1H), 6.89 (s, 1H), 4.48 (t, J = 5.4 Hz, 2H), 4.11 (t, J = 7.4 Hz, 4H), 3.99 (t, J = 7.17 Hz, 2H), 3.54 (m, 4H), 3.4 (m, 4H), 2.36 (m, 8H), 2.25 (m, 2H), 1.92 (m, 6H), 1.67 (m, 2H), 1.52 (m, 10H), 1.25 (m, 50H), 0.87 (m, 12H); CIMS m/z [M+H] + 990.1. Analytical HPLC column: Agilent Zorbax SB-C18, 5 μm, 4.6×150 mm, mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: ELSD, t R = 8.3 min, purity: > 99%; UPLC column: Waters Aquity UPLC® CSHTM, C18, 1.7 μm, 3.0×150 mm, (Part No. 186005302), mobile phase A: acetonitrile with 0.1% trifluoroacetic acid, mobile phase B: water with 0.1% trifluoroacetic acid, use gradient: A in B 5% to 95% in 15 min, flow rate: 1mL/min, column temperature: 20±2 °C, detector: CAD, t R = 13.4 min, purity: > 99 %. [1492] Preparation of Lipid Nanoparticles – General Procedure [1493] Representative Lipids of the Disclosure, distearoylphosphatidylcholine (DSPC), cholesterol, and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) were dissolved in pure ethanol at a molar ratio of 48.5:10:40:1.5 (IM administration) or 48.5:10:39:2.5 (IV administration) with a total lipid concentration of 10.8 mM. See, e.g., Qiu et al., PNAS 118:e2020401118 (2021). The lipid solution was then mixed with an acidic sodium acetate buffer (pH 4.0) or sodium citrate buffer (pH 4.0) containing mRNA (0.10 mg/mL0) by using the NanoAssemblr microfluidic system. The mRNA solution and the lipid solution were each injected into the NanoAssemblr microfluidic device at 12 mL/min total flow rate with mRNA solution to lipid solution at ratio of 3:1, and the device resulted in the rapid mixing of the two components and thus the self-assembly of LNPs. Formulations were further dialyzed against PBS (pH 7.4) or 20 mM Tris (pH 7.4) with 8% sucrose solution in dialysis cassettes overnight at 4 °C. The particle size of formulations was measured by dynamic light scattering (DLS) using a Zetasizer Ultra (Malvern Panalytical). RNA encapsulation efficiency was characterized by Ribogreen assay. Example 11. LNP Formulations A [1494] Ionizable lipids, DSPC, cholesterol, and PEG2K-DMG were dissolved in pure ethanol at a 48.5:10:39:2.5 mol% ratio with a total lipid concentration of 10.8 mM. A 0.10 mg/mL mRNA solution was prepared using acidic buffer (pH 4.0-5.0) containing mRNAs encoding human erythropoietin (hEPO) and firefly luciferase (fLuc) (1:2 ratio). The nucleotide and lipid solutions were mixed at a 3:1 volume ratio using the NanoAssemblr microfluidic system at a 12 mL/min total flow rate resulting in rapid mixing and self-assembly of LNPs. Formulations were further dialyzed against PBS (pH 7.4) overnight at 4 °C, concentrated using centrifugal filtration and filtered (0.2 µm pore size). The particle size and polydispersity index (PDI) of formulations was measured by dynamic light scattering (DLS) using a Zetasizer Ultra (Malvern Panalytical). RNA encapsulation efficiency (EE%) was determined by Ribogreen assay. Table 2A: LNP Formulations [1495] Buffer A: 25 mM Sodium Acetate, pH 5.0; Buffer B: 50 mM Citrate, pH 4.0 Example 12. LNP Formulations B [1496] Ionizable lipids, DSPC, cholesterol, and PEG2K-DSPE were dissolved in pure ethanol at a 48.5:10:40:1.5 mol% ratio with a total lipid concentration of 10.8 mM. A 0.10 mg/mL mRNA solution was prepared using acidic buffer (pH 4.0-5.0) containing mRNAs encoding firefly luciferase (fLuc). The nucleotide and lipid solutions were mixed at a 3:1 volume ratio using the NanoAssemblr microfluidic system at a 12 mL/min total flow rate resulting in rapid mixing and self-assembly of LNPs. Formulations were further dialyzed against PBS (pH 7.4) overnight at 4 °C, concentrated using centrifugal filtration and filtered (0.2 µm pore size). The particle size and polydispersity index (PDI) of formulations was measured by dynamic light scattering (DLS) using a Zetasizer Ultra (Malvern Panalytical). RNA encapsulation efficiency (EE%) was determined by Ribogreen assay. Table 2B: LNP Formulations [1497] Buffer A: 25 mM Sodium Acetate, pH 5.0; Buffer B: 50 mM Citrate, pH 4.0 Example 13. LNP Formulations C [1498] Ionizable lipids, DSPC, cholesterol, and PEG2K-DMG were dissolved in pure ethanol at a 48.5:10:40:1.5 mol% ratio (formulations comprising ionizable lipids of the present disclosure) or 50:10:38.5:1.5 mol% ratio (SM102 formulations) with a total lipid concentration of 10.8 mM. A 0.10 mg/mL RNA solution was prepared using acidic buffer (pH 4.0-5.0) containing circular RNAs (oRNA) or linear mRNA encoding COVID spike protein, as indicated in Table XC. The nucleotide and lipid solutions were mixed at a 3:1 volume ratio using the NanoAssemblr microfluidic system at a 12 mL/min total flow rate resulting in rapid mixing and self-assembly of LNPs. Formulations were further dialyzed against cryobuffer overnight at 4 °C, concentrated using centrifugal filtration and filtered (0.2 µm pore size). The formulations were then stored at -80°C until used. The particle size and polydispersity index (PDI) of formulations was measured by dynamic light scattering (DLS) using a Zetasizer Ultra (Malvern Panalytical). RNA encapsulation efficiency (EE%) was determined by Ribogreen assay. Table 2C: LNP Formulations Example 14. hEPO and fLUC in vivo reporter assays [1500] Balb/cAnNCrl (female, 6-8 weeks) were administrated with LNPs (formulated with 0.1 mg/kg EPO and 0.2 mg/kg Luc, see Example 11) by intravenous injection. Plasma samples were harvested at 5, 23 and 47 hours post dose for hEPO analysis. Bioluminescence imaging (BLI) of the mice was taken at 6, 24 and 48 hours post-dosing using an IVIS Lumina III LT system (PerkinElmer) after injection of D-luciferin solution (150 mg/kg, intraperitoneal injection (IP)) to determine. hEPO concentrations were measured using an ELISA kit (DEP00, R&D Systems). The maximal concentration or BLI signal (Cmax) and area under concentration vs time curve (AUC) of the individual mouse plasma hEPO or whole body BLI data was calculated using a non-compartment analysis (NCA) program (WinNonlin ® , Version 8.3.4 [Pharsight Corp (Mountain View, CA, USA)]). [1501] Table Y reports the hEPO concentration at 5 hours and the AUC over the 48 hour period after dosing, both overall and vs an internal standard across experiments, for each formulation tested. Table Y also reports the luciferase bioluminescence imaging measured at 6 hours and the AUC over the 48 hour period after dosing, for each formulation tested. [1502] Data keys: [1503] hEPO C5hr (IU/µL): + = <10 IU/µL; 10 IU/µL ≤ ++ < 100 IU/µL; 100 ≤ +++ < 1,000 IU/µL [1504] hEPO AUC (hr*IU/µL): + = <10 hr*IU/µL; 10 hr*IU/µL ≤ ++ < 100 hr*IU/µL; 100 hr*IU/µL ≤ +++ < 1,000 hr*IU/µL; 1,000 hr*IU/µL ≤ ++++ < 10,000 hr*IU/µL [1505] AUC ratio vs standard: * = < 0.1; 0.1 ≤ ** < 0.5; 0.5 ≤ *** < 1.0; 1.0 ≤ **** < 1.5; 1.5 ≤ ***** < 2.0; ****** ≥ 2.0 [1506] Luciferase BLI C6hr (photons/sec): # = <100 million p/s; 100 million p/s ≤ ## < 1 billion p/s; 1 billion p/s ≤ ### < 10 billion p/s; 10 billion p/s ≤ #### < 100 billion p/s; 100 billion p/s ≤ ##### < 1 trillion p/s [1507] Luciferase BLI AUC48hr (hr*photons/sec): $ < 10 billion hr*p/s; 10 billion hr*p/s ≤ $$ < 100 billion hr*p/s; 100 billion hr*p/s ≤ $$$ < 1 trillion hr*p/s; 1 trillion hr*p/s ≤ $$$$ < 10 trillion hr*p/s Table 3: In Vivo Assay Data Example 15. In Vivo Organ Tropism assays [1508] Balb/cAnNCrl (female, 6-8 weeks) were dosed LNP formulations (formulated with 0.2 mg/kg Luc mRNA, see Example 12) by IV injection. At 6 hour post LNP dose, the mice were injected with D-luciferin solution (150 mg/kg, intraperitoneal (IP)).10 minutes post D-luciferin dosing, Mice were sacrificed and organs (liver, spleen, lung, heart, kidney) were harvested. Bioluminescence imaging of the organs from each dosing groups were taken simultaneously using an IVIS Lumina III LT system (PerkinElmer). [1509] The sum of the bioluminescence of all organs from each individual mouse were summed as the total flux (photons/second). The percentage of bioluminescence of each individual organ was calculated to determine the organ tropism of the LNP formulations. [1510] Luciferase BLI C6hr (photons/sec): # = <100 million p/s; 100 million p/s ≤ ## < 1 billion p/s Table 4. Total Flux of organs and percentage flux in each organ Example 16. In Vivo T cell responses to spike protein encoding RNA - Murine Dosing Protocol [1511] LNP formulations were prepared as described in Example 13. Each formulation was injected in 5 BALB/c mice intramuscularly on day 0 and 21 with 0.02 mg/ml oRNA or linear mRNA encoding COVID spike protein in a total volume of 0.5 mL. Prior to dosing BALB/c mice were placed in a chamber prefilled with isoflurane at a flow rate of 0.4-0.8 liter/min until sedated so that no movement occurred during injection. The injection site was monitored for irritation after both doses. On day 35 all mice were humanely euthanized by CO2 inhalation and spleens were collected and stored on wet ice until processing. All in vivo experiments in this study were performed under the approved animal care guidelines. Analysis [1512] Spleens were harvested and manually dissociated into single cell suspensions by filtration using a 70µm filter (Miltenyi 130-098-462) and washed with 1x PBS (Fisher 10010049) containing 2mM EDTA (ThermoFisher 15575-020) and 0.5% BSA (Miltenyi 130- 091-376). Red blood cells were lysed using ACK Lysisg Buffer (ThermoFisher A1049201) and washed twice with 1x PBS + 2mM EDTA + 0.5% BSA. Following final wash, cells were resuspended in 1x PBS and counted (ViCell XR, Beckman Coulter 731196). Cells were resuspended in CTL Test Plus Medium (C.T.L. CTLTP-005) containing 1x GlutaMAX (ThermoFisher TP-050122) and 1x Pen/Strep (ThermoFisher 15-140-122) at appropriate concentrations and plated for downstream functional assays. [1513] ELISpot analysis was performed using the mouse IFN-γ ELISpotPLUS Kit (Mabtech 3321-4HST-10), according to the manufacturer’s protocol. Briefly, plates were washed with 1x PBS and blocked with RPMI (ThermoFisher 72400-047) containing 10% FBS (ThermoFisher A38400-01) for 1 h at 37°C. Following blocking, cells were plated at 200,000 cells/well for DMSO and peptide-stimulated wells or 25,000 cells/well for PMA/Ionomycin treatment. Cells were incubated with either 1% DMSO (ThermoFisher D12345), 7.5µg/mL of S1 or S2 peptide pools spanning the Spike protein of SARS-CoV-2 (JPT PM-WCPV-S-1), or 1x PMA/Ionomycin (ThermoFisher 00-4970-93) in triplicate. The plates were incubated overnight 37°C, 5% CO2. Following incubation, plates were washed, and 1 µg/mL detection antibody added for 2h at room temperature. Washes were repeated and 1x Streptavidin-HRP added and incubated for 1hr at room temperature. Finally, plates were washed and TMB substrate added, incubated in the dark for spot development, then washed out using tap water. Plates were allowed to dry and counted by an ELISpot analyzer (ZellNet Consulting). [1514] For intracellular staining (ICS) 5,000,000 cells per well were plated in a 96-well round bottom plate (Costar 3799) and stimulated using the same ELISpot conditions as described above and incubated at 37C with 5% CO2 for a total of 5.5h. Golgi Plug (BD 555029) was added to all wells for the last 4.5h of stimulation. Following incubation, cells were stained for flow cytometry using surface or intracellular antibodies listed in the table below. Briefly, cells were washed with 1x PBS and stained with Live/Dead Fixable Aqua (Invitrogen L34966) for 20min at room temperature. Cells were then washed twice with Cell Staining Buffer (BioLegend 420201) and incubated with Fc Block (Biolegend 156604) for 5min at 4°C, followed by surface antibody staining for 30min at 4°C. Thereafter, cells were washed twice with Cell Staining Buffer, fixed at 4°C for 30min IC Fixation Buffer (ThermoFisher 88-8824- 00) and permeabilized in 1x permeabilization buffer (ThermoFisher 88-8824-00) and intracellular staining performed overnight at 4°C.Thereafter, cells were washed twice with 1x permeabilization buffer, resuspended in 1x PBS, and acquired on cytometer (ThermoFisher Attune NXT with a laser configuration of Blue(3)/Red(3)/Violet(4)/Yellow(4)) equipped with a high-throughput autosampler (ThermoFisher CytKick). Compensation was performed using UltraComp eBeads (ThermoFisher 01-3333-41) and ArC Amine Reactive Compensation Bead Kit (ThermoFisher A10346). Results [1515] Mice dosed with LNP formulation F-16 demonstrated spike-specific polyfunctional CD4 T Cell responses comparable to the Moderna SM102 LNP formulations F-16 and F-17. Example 17. In Vivo T cell responses to spike protein encoding RNA – Non-human Primate Dosing Protocol [1516] LNP formulations were prepared as described in Example 13. Each formulation was injected in 3 non-naïve Cynomolgus monkeys once on Day 1 and once on Day 22 via intramuscular injections at a dose level of 100 µg. All NHPs were temporarily restrained for dose administration and not sedated. Prior to dosing, the dosing site was shaved and marked as necessary for clinical observations. Each dose was administered using a syringe/needle within the demarcated area. Samples were collected throughout the study for clinical pathology parameters, pharmacokinetic analysis and immunogenicity analysis. Analysis [1517] Cryopreserved PBMCs were thawed in a 37°C water bath and cells transferred to conical tube containing complete RPMI (RPMI [ThermoFisher 72400-047] containing 10% FBS [ThermoFisher A38400-01] and 1x Pen/Strep [ThermoFisher 15-140-122]. Cells were centrifuged, resuspended in complete RPMI containing 50U/mL Benzonase (EMD 70664- 10KUN), and incubated for 15min at 37°C. Cells were centrifuged, resuspended in complete RPMI, and rested for 3hr. Cells were centrifuged and resuspended in CTL Test Plus Medium (C.T.L. CTLTP-005) containing 1x GlutaMAX (ThermoFisher TP-050122) and 1x Pen/Strep and counted (ViCell XR, Beckman Coulter 731196). Concentrations were adjusted and cells plated for downstream functional assays. [1518] ELISpot analysis was performed using the Monkey IFN-γ ELISpotPLUS Kit (Mabtech 3421M-4HST-10) according to the manufacturer’s protocol. Briefly, plates were washed with 1x PBS and blocked with RPMI (ThermoFisher 72400-047) containing 10% FBS (ThermoFisher A38400-01) for 1h at 37°C. Following blocking, cells were plated in triplicate and stimulated under the following conditions: no peptide (1% DMSO (ThermoFisher D12345)),0.5µg/mL of S1+S2 peptide pools spanning the Spike protein of SARS-CoV-2 (JPT PM-WCPV-S-1), and 1x PMA/Ionomycin (ThermoFisher 00-4970-93). 200,000 cells/well were plated for DMSO and peptide pool stimulations and 10,000/well from pooled samples from each group for PMA/Ionomycin stimulation. [1519] The plates were incubated overnight 37°C, 5% CO2. Following incubation plates were washed, and 1 µg/mL detection antibody added for 2h at room temperature. Washes were repeated and 1x Streptavidin-HRP added and incubated for 1h at room temperature. Finally, plates were washed and TMB substrate added, incubated in the dark for spot development, then washed out using tap water. Plates were allowed to dry and counted using an ELISpot analyzer (ZellNet Consulting). [1520] For intracellular staining (ICS), approximately 2,000,000 cells per well from each animal were plated in a 96-well round bottom plate (Costar 3799) and stimulated using the same ELISpot conditions described above. After one hour of stimulation, Golgi Plug (BD 555029) was added to all wells and plates incubated overnight at 37°C with 5% CO 2 . Thereafter, cells were washed and stained for flow cytometry. Briefly, cells were washed with 1x PBS and stained with Live/Dead Fixable Aqua (Invitrogen L34966) for 20min at room temperature. Cells were then washed twice with Cell Staining Buffer (BioLegend 420201) and incubated with Fc Block (Biolegend 156604) for 5min at 4°C, followed by surface antibody staining for 30min at 4°C. Following surface staining, cells were then washed twice with Cell Staining Buffer and fixed at 4°C for 30min in IC Fixation Buffer and permeabilized in (ThermoFisher 88-8824-00) 1x permeabilization buffer (ThermoFisher 88-8824-00). Intracellular staining was performed for 1hr at 4°C. Thereafter, cells were washed twice with 1x permeabilization buffer, resuspended in 1x PBS, and acquired on cytometer (ThermoFisher Attune NXT with a laser configuration of Blue(3)/Red(3)/Violet(4)/Yellow(4)) equipped with a high-throughput autosampler (ThermoFisher CytKick). Compensation was performed using UltraComp eBeads (ThermoFisher 01-3333-41) and ArC Amine Reactive Compensation Bead Kit (ThermoFisher A10346). Results [1521] NHPs dosed with LNP formulation F-16 demonstrated spike-specific polyfunctional CD4 T Cell responses that were greater than or comparable to the Moderna SM102 LNP formulations as determined by ICS assay. At day 36, formulation F-16 demonstrated ~10 fold greater spike-specific T Cell response as compared to control formulation F-17, and similar response to control formulation F-18. At Day 36, T cells from NHPs dosed with F-16 also demonstrated increased spike-specific IFN ^ secreting T cells that were 3 fold higher than those induced by F-17 and comparable to F-18 formulations as determined via ELISpot assay. XIII. EQUIVALENTS AND SCOPE [1522] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments in accordance with the disclosure described herein. The scope of the present disclosure is not intended to be limited to the above Description, but rather is as set forth in the appended claims. [1523] In the claims, articles such as "a," "an," and "the" may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include "or" between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The disclosure includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The disclosure includes embodiments in which more than one, or the entire group members are present in, employed in, or otherwise relevant to a given product or process. [1524] It is also noted that the term "comprising" is intended to be open and permits but does not require the inclusion of additional elements or steps. When the term "comprising" is used herein, the term "consisting of" is thus also encompassed and disclosed. [1525] The term "about" as used herein, means within 10% of a given value or range. Thus, "about 10" means 9 to 11. [1526] Where ranges are given, endpoints are included. Furthermore, it is to be understood that unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or subrange within the stated ranges in different embodiments of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. [1527] In addition, it is to be understood that any particular embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Since such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the compositions of the disclosure (e.g., any antibiotic, therapeutic or active ingredient; any method of production; any method of use; etc.) can be excluded from any one or more claims, for any reason, whether or not related to the existence of prior art. [1528] It is to be understood that the words which have been used are words of description rather than limitation, and that changes may be made within the purview of the appended claims without departing from the true scope and spirit of the disclosure in its broader aspects. [1529] While the present disclosure has been described at some length and with some particularity with respect to the several described embodiments, it is not intended that it should be limited to any such particulars or embodiments or any particular embodiment, but it is to be construed with references to the appended claims so as to provide the broadest possible interpretation of such claims in view of the prior art and, therefore, to effectively encompass the intended scope of the disclosure.
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