HAMILTON GREGORY LAWRENCE (US)
WO2003101952A2 | 2003-12-11 | |||
WO2015095340A1 | 2015-06-25 | |||
WO2020152037A1 | 2020-07-30 |
EP1164125A1 | 2001-12-19 | |||
EP3842538A1 | 2021-06-30 | |||
JP2005181734A | 2005-07-07 | |||
US8058069B2 | 2011-11-15 | |||
US9364435B2 | 2016-06-14 | |||
US10113163B2 | 2018-10-30 | |||
US20210130805A1 | 2021-05-06 | |||
US10167457B2 | 2019-01-01 | |||
US9840699B2 | 2017-12-12 | |||
US20030083272A1 | 2003-05-01 | |||
US20170210698A1 | 2017-07-27 | |||
US20180058171W | 2018-10-30 |
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Claims We claim: 1. A compound of formula I: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein: each of L1, L1’, and L1” is independently absent, -C(O)-, or -OC(O)-; each of L2, L2’, and L2” is independently absent, an optionally substituted bivalent saturated or unsaturated, straight or branched C1-12 hydrocarbon chain, or –(CH2)m-CyA-(CH2)m’-; each CyA is independently an optionally substituted ring selected from phenylene and 3- to 12- membered saturated or partially unsaturated carbocyclylene; each of m and m’ is independently 0, 1, or 2; each of L3, L3’, and L3” is independently absent, -C(O)-, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-; each of R1, R1’, and R1” is independently – ( )p y , , or an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain wherein 1-3 methylene units are optionally and independently replaced with –O- or –NR-; each CyB is independently an optionally substituted ring selected from 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl; each p is independently 0, 1, 2, or 3; each L3a is independently absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C1-10 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O- or -NR-; each Ra is independently hydrogen or an optionally substituted group selected from C6-20 aliphatic, 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl; X1 is absent, –O–, –S–, or –NR–; X2 is absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –CyC-; CyC is an optionally substituted ring selected from 3- to 7- membered saturated or partially unsaturated carbocyclene, phenylene, 3- to 7-membered saturated or partially unsaturated heterocyclene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or 5- to 6-membered heteroarylene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; X3 is hydrogen or an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl, phenyl, 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and each R is independently hydrogen or an optionally substituted C1-6 aliphatic group. 2. The compound of claim 1, wherein the compound is of Formula I-A: or its N-oxide, or a pharmaceutically acceptable salt thereof. 3. The compound of claims 1 or 2, wherein the compound is of Formula I-B: or its N-oxide, or a pharmaceutically acceptable salt thereof. 4. The compound of any one of the preceding claims, wherein the compound is of Formula I-C: or its N-oxide, or a pharmaceutically acceptable salt thereof. 5. The compound of any one of the preceding claims, wherein the compound is of Formula I-D: or its N-oxide, or a pharmaceutically acceptable salt thereof. 6. The compound of any one of the preceding claims, wherein the compound is of Formula I-E: or its N-oxide, or a pharmaceutically acceptable salt thereof. 7. The compound of claim 1, wherein L1 is -C(O)-. 8. The compound of claim 1, wherein L1’ is -C(O)-. 9. The compound of claim 1, wherein L1” is -C(O)-. 10. The compound of any one of claims 1-9, wherein L2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C1-12 hydrocarbon chain. 11. The compound of any one of claims 1-10, wherein L2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C1-12 hydrocarbon chain. 12. The compound of any one of claims 1-11, wherein L2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C1-12 hydrocarbon chain. 13. The compound of any one of claims 1-9 or 11-12, wherein L2 is absent. 14. The compound of any one of claims 1-10 or 12-13, wherein L2’ is absent. 15. The compound of any one of claims 1-11 or 13-14, wherein L2” is absent. 16. The compound of any one of claims 1-15, wherein L3 is absent. 17. The compound of any one of claims 1-16, wherein L3’ is absent. 18. The compound of any one of claims 1-17, wherein L3” is absent. 19. The compound of any one of claims 1-15, wherein L3 is -C(O)O- or –OC(O)-. 20. The compound of any one of claims 1-16 and 18-19, wherein L3’ is -C(O)O- or –OC(O)-. 21. The compound of any one of claims 1-17 and 19-20, wherein L3” is -C(O)O- or –OC(O)-. 22. The compound of any one of claims 1-21, wherein R1 is -(CH2)p-CyB. 23. The compound of any one of claims 1-21, wherein R1 is . 24. The compound of any one of claims 1-21, wherein R1 is an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain. 25. The compound of any one of claims 1-24, wherein R1’ is -(CH2)p-CyB. 26. The compound of any one of claims 1-24, wherein R1’ is . 27. The compound of any one of claims 1-24, wherein R1’ is an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain. 28. The compound of any one of claims 1-27, wherein R1” is -(CH2)p-CyB. 29. The compound of any one of claims 1-27, wherein 30. The compound of any one of claims 1-27, wherein R1” is an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain. 31. The compound of any one of claims 1-30, wherein each CyB is 1-adamantyl. 32. The compound of any one of claims 1-31, wherein each p is independently 0 or 1. 33. The compound of any one of claims 1-32, wherein X1 is –O–. 34. The compound of any one of claims 1-32, wherein X1 is –NR–. 35. The compound of any one of claims 1-34, wherein X2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –CyC-. 36. The compound of any one of claims 1-35, wherein X2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C4-8 hydrocarbon chain, wherein 1 methylene unit is replaced with -NR-. 37. The compound of any one of claims 1-35, wherein X2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C4-8 hydrocarbon chain, wherein 1 methylene unit is replaced with –CyC-. 38. The compound of any one of claims 1-37, wherein X3 is hydrogen. 39. The compound of any one of claims 1-36, wherein X3 is optionally substituted 3- to 7- membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. 40. The compound of any one of the preceding claims, wherein -X2-X3 is , 41. The compound of any one of claims 1-40, R is hydrogen. 42. The compound of any one of claims 1-40, R is optionally substituted C1-6 aliphatic. 43. A compound selected from Table 1, or a pharmaceutically acceptable salt thereof. 44. A lipid nanoparticle (LNP) preparation comprising an ionizable lipid of any one of claims 1-43. 45. A lipid nanoparticle (LNP) preparation comprising: an ionizable lipid of any one of claims 1-43; a phospholipid; a sterol; and a conjugate-linker lipid (e.g., polyethylene glycol lipid). 46. The LNP preparation of claim 44, further comprising a therapeutic and/or prophylactic agent. 47. The LNP preparation of claim 46, wherein the therapeutic and/or prophylactic agent is or comprises one or more nucleic acids. 48. The LNP preparation of claim 47, wherein the one or more nucleic acids is or comprises RNA. 49. The LNP preparation of claim 47, wherein the one or more nucleic acids is or comprises DNA. 50. The LNP preparation of any one of claims 46-49, wherein the LNP preparation is formulated to deliver the therapeutic and/or prophylactic agent to target cells. 51. The LNP preparation of claim 50, wherein the target cells are or comprise spleen cells (e.g., splenic B cells, splenic T cells, splenic monocytes), liver cells (e.g., hepatocytes), bone marrow cells (e.g., bone marrow monocytes), immune cells, kidney cells, muscle cells, heart cells, lung cells, or cells in the central nervous system. 52. The LNP preparation of claim 51, wherein the target cells are or comprise hematopoietic stem cells (HSCs). 53. A pharmaceutical composition comprising a LNP preparation of any one of claims 44-52 and a pharmaceutically acceptable excipient. 54. A method for administering a therapeutic and/or prophylactic agent to a subject in need thereof, the method comprising administering the LNP preparation of any one of claims 44-52 or the pharmaceutical composition of claim 53 to the subject. 55. A method for treating a disease or a disorder in a subject in need thereof, the method comprising administering the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53, to the subject, wherein the therapeutic and/or prophylactic agent is effective to treat the disease. 56. A method for delaying and/or arresting progression a disease or a disorder in a subject in need thereof, the method comprising administering the LNP preparation of any one of claims 44- 52, or the pharmaceutical composition of claim 53, to the subject, wherein the therapeutic and/or prophylactic agent is effective to treat the disease. 57. A method of delivering a therapeutic and/or prophylactic agent to a mammalian cell derived from a subject, the method comprising contacting the cell of the subject having been administered the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53. 58. A method of producing a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53, wherein the therapeutic and/or prophylactic agent is or comprises an mRNA, and wherein the mRNA encodes the polypeptide of interest, whereby the mRNA is capable of being translated in the cell to produce the polypeptide of interest. 59. A method of inhibiting production of a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53, wherein the therapeutic and/or prophylactic agent is or comprises an RNA, whereby the RNA is capable of inhibiting production of the polypeptide of interest. 60. A method of specifically delivering a therapeutic and/or prophylactic agent to a mammalian organ, tissue, cell, or population of cells, the method comprising contacting a mammalian organ, tissue, cell, or population of cells with the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53, whereby the therapeutic and/or prophylactic agent is delivered to the organ, tissue, cell, or population of cells. 61. The method of claim 60, comprising administering to a subject the LNP preparation of any one of claims 44-52, or the pharmaceutical composition of claim 53, to the subject. 62. A method of vaccinating by administering the LNP preparation of any one of claims 44- 52, or the pharmaceutical composition of claim 53. 63. A method of inducing an adaptive immune response in a subject, comprising administering to the subject an effective amount of a composition comprising at least one RNA; wherein the composition comprises a LNP preparation comprising a compound of any one of claims 1-43, or a pharmaceutically acceptable salt thereof. |
or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , L 3’ , L 3” , R 1 , R 1’ , R 1” , X 1 , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0105] In some embodiments, the present disclosure provides a compound of Formula I-A-a: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , L 3’ , L 3” , R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0106] In some embodiments, the present disclosure provides a compound of Formula I-A-b: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , L 3’ , L 3” , R, R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0107] In some embodiments, the present disclosure provides a compound of Formula I-A-c:
or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , L 3’ , L 3” , R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0108] In some embodiments, the present disclosure provides a compound of Formula I-B: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , R 1 , R 1’ , R 1” , X 1 , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0109] In some embodiments, the present disclosure provides a compound of Formula I-B-a: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0110] In some embodiments, the present disclosure provides a compound of Formula I-B-b: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , R, R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0111] In some embodiments, the present disclosure provides a compound of Formula I-B-c: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , R 1 , R 1’ , R 1” , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0112] In some embodiments, the present disclosure provides a compound of Formula I-C: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , R 1 , R 1’ , R 1” , X 1 , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0113] In some embodiments, the present disclosure provides a compound of Formula I-D: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 2’ , L 2” , L 3 , R 1 , R 1’ , R 1” , X 1 , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0114] In some embodiments, the present disclosure provides a compound of Formula I-E: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein each of L 2 , L 3 , R 1 , R 1’ , R 1” , X 1 , X 2 , and X 3 is as described above and in classes and subclasses herein, both singly and in combination. [0115] In some embodiments of any Formulae described herein, L 1 is absent, -C(O)-, or -OC(O)- . In some embodiments, L 1 is absent. In some embodiments, L 1 is -C(O)-, or -OC(O)-. In some embodiments, L 1 is -C(O)-. In some embodiments, L 1 is -OC(O)-. It will be appreciated that when L 1 is –OC(O)-, the carbonyl of L 1 is attached to the oxygen depicted in Formula I and the oxygen of L 1 is attached to L 2 , such that a carbonate moiety is formed (e.g., -O-C(O)-O-L 2 -). [0116] In some embodiments of any Formulae described herein, L 1’ is absent, -C(O)-, or -OC(O)- . In some embodiments, L 1’ is absent. In some embodiments, L 1’ is -C(O)-, or -OC(O)-. In some embodiments, L 1’ is -C(O)-. In some embodiments, L 1’ is -OC(O)-. It will be appreciated that when L 1’ is –OC(O)-, the carbonyl of L 1’ is attached to the oxygen depicted in Formula I and the oxygen of L 1’ is attached to L 2’ , such that a carbonate moiety is formed (e.g., -O-C(O)-O-L 2’ -). [0117] In some embodiments of any Formulae described herein, L 1” is absent, -C(O)-, or -OC(O)- . In some embodiments, L 1” is absent. In some embodiments, L 1” is -C(O)-, or -OC(O)-. In some embodiments, L 1” is -C(O)-. In some embodiments, L 1” is -OC(O)-. It will be appreciated that when L 1” is –OC(O)-, the carbonyl of L 1” is attached to the oxygen depicted in Formula I and the oxygen of L 1” is attached to L 2” , such that a carbonate moiety is formed (e.g., -O-C(O)-O-L 2” -). [0118] In some embodiments, L 1 , L 1’ , and L 1” are the same. In some embodiments, L 1 and L 1’ are the same, which are different from L 1” . In some embodiments, L 1’ and L 1” are the same, which are different from L 1 . In some embodiments, L 1 and L 1” are the same, which are different from L 1’ . In some embodiments, L 1 , L 1’ , and L 1” are different from each other. [0119] In some embodiments of any Formulae described herein, L 2 is absent, an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain, or – (CH 2 )m-Cy A -(CH 2 )m’-. In some embodiments, L 2 is absent. In some embodiments, L 2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2 is –(CH 2 )m-Cy A -(CH 2 )m’-. In some embodiments, L 2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C1-8 hydrocarbon chain. In some embodiments, L 2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2 is an optionally substituted bivalent saturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2 is an optionally substituted bivalent saturated, straight or branched C 1 - 8 hydrocarbon chain. In some embodiments, L 2 is an optionally substituted bivalent saturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2 is -CH 2 -, -(CH 2 ) 2 -, -(CH 2 ) 3 -, -(CH 2 ) 4 -, -(CH 2 ) 5 -, - (CH 2 ) 6 -, or -(CH 2 ) 7 -. In some embodiments, L 2 is -CH 2 -. In some embodiments, L 2 is -(CH 2 ) 2 -. In some embodiments, L 2 is -(CH 2 ) 3 -. In some embodiments, L 2 is -(CH 2 ) 4 -. In some embodiments, L 2 is -(CH 2 ) 5 -. In some embodiments, L 2 is -(CH 2 ) 6 -. In some embodiments, L 2 is -(CH 2 )7-. [0120] In some embodiments of any Formulae described herein, L 2’ is absent, an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain, or – (CH 2 )m-Cy A -(CH 2 )m’-. In some embodiments, L 2’ is absent. In some embodiments, L 2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2’ is –(CH 2 ) m -Cy A -(CH 2 ) m’ -. In some embodiments, L 2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1 - 8 hydrocarbon chain. In some embodiments, L 2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2’ is an optionally substituted bivalent saturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2’ is an optionally substituted bivalent saturated, straight or branched C1-8 hydrocarbon chain. In some embodiments, L 2’ is an optionally substituted bivalent saturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2’ is -CH 2 -, -(CH 2 ) 2 -, - (CH 2 ) 3 -, -(CH 2 ) 4 -, -(CH 2 ) 5 -, -(CH 2 ) 6 -, or -(CH 2 ) 7 -. In some embodiments, L 2’ is -CH 2 -. In some embodiments, L 2’ is -(CH 2 )2-. In some embodiments, L 2’ is -(CH 2 )3-. In some embodiments, L 2’ is -(CH 2 ) 4 -. In some embodiments, L 2’ is -(CH 2 ) 5 -. In some embodiments, L 2’ is -(CH 2 ) 6 -. In some embodiments, L 2’ is -(CH 2 ) 7 -. [0121] In some embodiments of any Formulae described herein, L 2” is absent, an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain, or – (CH 2 ) m -Cy A -(CH 2 ) m’ -. In some embodiments, L 2” is absent. In some embodiments, L 2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2” is –(CH 2 )m-Cy A -(CH 2 )m’-. In some embodiments, L 2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1 - 8 hydrocarbon chain. In some embodiments, L 2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2” is an optionally substituted bivalent saturated, straight or branched C 1-12 hydrocarbon chain. In some embodiments, L 2” is an optionally substituted bivalent saturated, straight or branched C 1 - 8 hydrocarbon chain. In some embodiments, L 2” is an optionally substituted bivalent saturated, straight or branched C 1-6 hydrocarbon chain. In some embodiments, L 2” is -CH 2 -, -(CH 2 ) 2 -, - (CH 2 ) 3 -, -(CH 2 ) 4 -, -(CH 2 ) 5 -, -(CH 2 ) 6 -, or -(CH 2 ) 7 -. In some embodiments, L 2’’ is -CH 2 -. In some embodiments, L 2’’ is -(CH 2 )2-. In some embodiments, L 2’’ is -(CH 2 )3-. In some embodiments, L 2’’ is -(CH 2 )4-. In some embodiments, L 2’’ is -(CH 2 ) 5 -. In some embodiments, L 2’’ is -(CH 2 ) 6 -. In some embodiments, L 2’’ is -(CH 2 ) 7 -. [0122] In some embodiments, L 2 , L 2’ , and L 2” are the same. In some embodiments, L 2 and L 2’ are the same, which are different from L 2” . In some embodiments, L 2’ and L 2” are the same, which are different from L 2 . In some embodiments, L 2 and L 2” are the same, which are different from L 2’ . In some embodiments, L 2 , L 2’ , and L 2” are different from each other. [0123] In some embodiments of any Formulae described herein, each Cy A is independently an optionally substituted ring selected from phenylene and 3- to 12-membered saturated or partially unsaturated carbocyclylene. In some embodiments, Cy A is phenylene. In some embodiments, Cy A is 3- to 12-membered saturated or partially unsaturated carbocyclylene. [0124] In some embodiments of any Formulae described herein, each m is independently 0, 1, or 2. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. [0125] In some embodiments of any Formulae described herein, each m’ is independently 0, 1, or 2. In some embodiments, m’ is 0. In some embodiments, m’ is 1. In some embodiments, m’ is 2. [0126] In some embodiments, m and m’ are the same. In some embodiments, m and m’ are different. [0127] In some embodiments of any Formulae described herein, L 3 is absent, -C(O)-, -C(O)O-, - OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3 is absent. In some embodiments, L 3 is - C(O)-, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3 is -C(O)-, -C(O)O-, - OC(O)-, or -OC(O)O-. In some embodiments, L 3 is -C(O)O- or -OC(O)-. In some embodiments, L 3 is -C(O)-. In some embodiments, L 3 is -C(O)O-. In some embodiments, L 3 is -OC(O)-. In some embodiments, L 3 is -O-. In some embodiments, L 3 is -OC(O)O-. [0128] In some embodiments of any Formulae described herein, L 3’ is absent, -C(O)-, -C(O)O-, - OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3’ is absent. In some embodiments, L 3’ is - C(O)-, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3’ is -C(O)-, -C(O)O-, - OC(O)-, or -OC(O)O-. In some embodiments, L 3’ is -C(O)O- or -OC(O)-. In some embodiments, L 3’ is -C(O)-. In some embodiments, L 3’ is -C(O)O-. In some embodiments, L 3’ is -OC(O)-. In some embodiments, L 3’ is -O-. In some embodiments, L 3’ is -OC(O)O-. [0129] In some embodiments of any Formulae described herein, L 3” is absent, -C(O)-, -C(O)O-, - OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3” is absent. In some embodiments, L 3” is - C(O)-, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-. In some embodiments, L 3” is -C(O)-, -C(O)O-, - OC(O)-, or -OC(O)O-. In some embodiments, L 3” is -C(O)O- or -OC(O)-. In some embodiments, L 3” is -C(O)-. In some embodiments, L 3” is -C(O)O-. In some embodiments, L 3” is -OC(O)-. In some embodiments, L 3” is -O-. In some embodiments, L 3” is -OC(O)O-. [0130] In some embodiments, L 3 , L 3’ , and L 3” are the same. In some embodiments, L 3 and L 3’ are the same, which are different from L 3” . In some embodiments, L 3’ and L 3” are the same, which are different from L 3 . In some embodiments, L 3 and L 3” are the same, which are different from L 3’ . In some embodiments, L 3 , L 3’ , and L 3” are different from each other. [0131] In some embodiments of any Formulae described herein , or an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain wherein 1-3 methylene units are optionally and independently replaced with –O- or –NR-. In some embodiments, R 1 is -(CH 2 )p-Cy B . In some embodiments . In some embodiments, R 1 is an optionally substituted saturated or unsaturated, straight or branched C 1-20 hydrocarbon chain. In some embodiments, R 1 is an optionally substituted saturated or unsaturated, straight or branched C6-20 hydrocarbon chain. In some embodiments, R 1 is an optionally substituted saturated or unsaturated, straight or branched C 6-12 hydrocarbon chain. In some embodiments, R 1 is an optionally substituted saturated or unsaturated, straight or branched C 8-9 In some embodiments, R 1 is . In some embodiments, R 1 is . In some embodiments, R 1 is In 1 some embodiments, R is [0132] In some embodiments of any Formulae described herein, , or an optionally substituted saturated or unsaturated, straight or branched C 1-20 hydrocarbon chain wherein 1-3 methylene units are optionally and independently replaced with –O- or –NR-. In some embodiments, R 1’ is -(CH 2 ) p -Cy B . In some embodiments, R 1’ is . In some embodiments, R 1’ is an optionally substituted saturated or unsaturated, straight or branched C 1-20 hydrocarbon chain. In some embodiments, R 1’ is an optionally substituted saturated or unsaturated, straight or branched C 6-20 hydrocarbon chain. In some embodiments, R 1’ is an optionally substituted saturated or unsaturated, straight or branched C 6-12 hydrocarbon chain. In some embodiments, R 1’ is an optionally substituted saturated or unsaturated, straight or branched embodiments, In some embodiments, R 1’ is . In some embodiments, R 1’ is . In some embodiments, R 1’ is . In some embodiments, R 1’ is . In some embodiments, R 1’ is . In some embodiments, R 1’ is . In some embodiments, R 1’ is . [0133] In some embodiments of any Formulae described herein, , or an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain wherein 1-3 methylene units are optionally and independently replaced with –O- or –NR-. In some embodiments, R 1” is -(CH 2 ) p -Cy B . In some embodiments, In some embodiments, R 1” is an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain. In some embodiments, R 1” is an optionally substituted saturated or unsaturated, straight or branched C6-20 hydrocarbon chain. In some embodiments, R 1” is an optionally substituted saturated or unsaturated, straight or branched C 6-12 hydrocarbon chain. In some embodiments, R 1” is an optionally substituted saturated or unsaturated, straight or branched C 8-9 hydrocarbon chain. In some embodiments, R 1” is , , . In some embodiments, In some embodiments, R is . In some embodiments, R 1’’ is . In some embodiments, . In some embodiments, R 1’’ is . In some embodiments, R 1’’ is . In some embodiments, R 1’’ is . In some embodiments, R 1’’ is . [0134] In some embodiments, R 1 , R 1’ , and R 1” are the same. In some embodiments, R 1 and R 1’ are the same, which are different from R 1” . In some embodiments, R 1’ and R 1” are the same, which are different from R 1 . In some embodiments, R 1 and R 1” are the same, which are different from R 1’ . In some embodiments, R 1 , R 1’ , and R 1” are different from each other. [0135] In some embodiments of any Formulae described herein, each Cy B is independently an optionally substituted ring selected from 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl. In some embodiments, Cy B is 1-adamantyl. [0136] In some embodiments of any Formulae described herein, each p is independently 0, 1, 2, or 3. In some embodiments, p is 0. In some embodiments, p is 1, 2, or 3. In some embodiments, p is 0 or 1. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. [0137] In some embodiments of any Formulae described herein, each L 3a is independently absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-10 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with - O- or -NR-. In some embodiments, L 3a is absent. In some embodiments, L 3a is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-10 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O- or -NR-. In some embodiments, L 3a is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C1-10 hydrocarbon chain. [0138] In some embodiments of any Formulae described herein, each R a is independently hydrogen or an optionally substituted group selected from C6-20 aliphatic, 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl. In some embodiments, R a is optionally substituted C6-20 aliphatic. In some embodiments, R a is optionally substituted C6-12 aliphatic. In some embodiments, R a is optionally substituted C 6-20 alkenyl. In some embodiments, R a is optionally substituted C 6-12 alkenyl. In some embodiments, R a is . [0139] In some embodiments, each [0140] In some embodiments of any Formulae described herein, X 1 is absent, –O–, –S–, or –NR– . In some embodiments, X 1 is absent. In some embodiments, X 1 is –O–, –S–, or –NR–. In some embodiments, X 1 is –O–. In some embodiments, X 1 is –NR–. In some embodiments, X 1 is –S–. [0141] In some embodiments of any Formulae described herein, X 2 is absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –Cy C -. In some embodiments, X 2 is absent. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –Cy C -. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -NR-. In some embodiments, X 2 is an optionally substituted, bivalent saturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -NR-. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1 methylene unit is replaced with -NR-. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 4-8 hydrocarbon chain, wherein 1 methylene unit is replaced with -NR-. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with –Cy C -. In some embodiments, X 2 is an optionally substituted, bivalent saturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with –Cy C -. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1 methylene unit is replaced with –Cy C -. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 4-8 hydrocarbon chain, wherein 1 methylene unit is replaced with –Cy C -. In some embodiments, X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain. In some embodiments, X 2 is an optionally substituted, bivalent saturated, straight or branched, C 1-12 hydrocarbon chain. In some embodiments, X 2 is an optionally substituted, bivalent saturated, straight or branched, C 1-6 hydrocarbon chain. In some embodiments, X 2 is an optionally substituted, bivalent saturated, straight or branched, C 2-3 hydrocarbon chain. [0142] In some embodiments of any Formulae described herein, Cy C is an optionally substituted ring selected from 3- to 7- membered saturated or partially unsaturated carbocyclene, phenylene, 3- to 7-membered saturated or partially unsaturated heterocyclene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or 5- to 6-membered heteroarylene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy C is optionally substituted 3- to 7-membered saturated or partially unsaturated heterocyclene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy C is optionally substituted 5- to 6-membered saturated or partially unsaturated heterocyclene having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy C is optionally substituted 5- to 6-membered saturated heterocyclene having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy C is 5- to 6-membered saturated heterocyclene having 1 nitrogen. In some embodiments, Cy C is an optionally substituted piperidine ring. In some embodiments, Cy C is a piperidine ring. [0143] In some embodiments of any Formulae described herein, X 3 is hydrogen or an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl, phenyl, 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 3 is hydrogen. In some embodiments, X 3 is optionally substituted 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 3 is optionally substituted 5- to 6-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 3 is optionally substituted 5- to 6-membered saturated heterocyclyl having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, X 3 is optionally substituted 5-membered saturated or partially unsaturated heterocyclyl having 1-2 nitrogen atoms. In some embodiments, X 3 is pyrrolidinyl. In some embodiments, X 3 is optionally substituted 6-membered saturated or partially unsaturated heterocyclyl having 1-2 nitrogen atoms. In some embodiments, X 3 is optionally substituted piperidinyl (e.g., piperidinyl substituted with C 1-6 alkyl). [0144] In some embodiments, In some embodiments, -X 2 -X 3 is . In some embodiments, -X 2 -X 3 is . In some embodiments, -X 2 -X 3 is . In some embodiments, -X 2 -X 3 is . [0145] In some embodiments of any Formulae described herein, each R is independently hydrogen or an optionally substituted C 1-6 aliphatic group. In some embodiments, R is hydrogen. In some embodiments, R is optionally substituted C 1-6 aliphatic. In some embodiments, R is optionally substituted C1-3 aliphatic. In some embodiments, R is –CH3 or -CH 2 CH3. [0146] It will be appreciated that “[compound/formula] or its N-oxide, or a pharmaceutically acceptable salt thereof”, as used herein, refers to pharmaceutically acceptable salts of i) the respective compound or formula or ii) N-oxides of such compound or formula. [0147] In some embodiments, the present disclosure provides a compound selected from Table 1. Table 1.
yyy
or a pharmaceutically acceptable salt thereof. [0148] It will be understood that, unless otherwise specified or prohibited by the foregoing definition of any of Formulae I, I-A, I-A-a, I-A-b, I-A-c, I-B, I-B-a, I-B-b, I-B-c, I-C, I-D, and I- E embodiments of variables L 1 , L 1’ , L 1” , L 2 , L 2’ , L 2” , L 3 , L 3’ , L 3” , R 1 , R 1’ , R 1” , Cy A , Cy B , Cy C , m, m’, p, L 3a , R a , R, X 1 , X 2 , and X 3 as defined above and described in classes and subclasses herein, apply to compounds of any of Formulae I, I-A, I-A-a, I-A-b, I-A-c, I-B, I-B-a, I-B-b, I-B-c, I-C, I- D, and I-E both singly and in combination. [0149] In some embodiments, provided compounds are compounds of any of Formulae I, I-A, I- A-a, I-A-b, I-A-c, I-B, I-B-a, I-B-b, I-B-c, I-C, I-D, and I-E or a pharmaceutically acceptable salt thereof. [0150] In some embodiments, provided compounds are provided and/or utilized in a salt form (e.g., a pharmaceutically acceptable salt form). Reference to a compound provided herein is understood to include reference to salts thereof, unless otherwise indicated. [0151] It will be appreciated that throughout the present disclosure, unless otherwise indicated, reference to a compound of Formula I is intended to also include compounds of any of Formulae I-A, I-A-a, I-A-b, I-A-c, I-B, I-B-a, I-B-b, I-B-c, I-C, I-D, and I-E and compound species of such formula disclosed herein. [0152] In some embodiments, the present disclosure encompasses the recognition that provided compounds display certain desirable characteristics, e.g., as compared to reference compounds or other known compounds. For example, in some embodiments, provided compounds exhibit more potent delivery to various cell types in one or more experiments described herein, and/or have one or more other characteristics that make them more suitable for delivery of cargos such as therapeutic or prophylactic agents than other known compounds. Without wishing to be bound by any particular theory, the present disclosure encompasses the recognition that provided compounds comprising a tetravalent core (e.g., a core derived from 2,2-bis(hydroxymethyl)propane-1,3-diol) feature display certain more desirable characteristics (e.g., more potent delivery to various cell types in one or more experiments described herein) than corresponding compounds lacking the same tetravalent core feature. B. Preparing Provided Compounds [0153] Provided compounds may generally be made by the processes described in the ensuing schemes and examples. C. Ionizable lipids [0154] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that comprise one or more ionizable lipids as described herein. [0155] Among other things, it was surprisingly found that different ratios of ionizable lipids influence one or more functional activities such as desired tropisms, stabilization, and drug delivery efficacy of compositions, preparations, nanoparticles, and/or nanomaterials described herein. For example, the present disclosure demonstrates a surprising finding that amounts of ionizable lipids different to those amounts described in the art (e.g., see U.S. Patent No.8,058,069 B2, or see, e.g., U.S. Patent No.9,364,435, the contents of both which are hereby incorporated by reference in their entireties herein) are important to and/or influence one or more functional activities of compositions, preparations, nanoparticles, and/or nanomaterials described herein. For example, in some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials having an ionizable lipid that is at about 50 mol percent or less, based on total moles of components of the lipid nanoparticle, was found to be useful and/or critical to functional activity of lipid nanoparticles such as desired tropisms, stabilization, and drug delivery efficacy as described herein. [0156] In some embodiments, an ionizable lipid may include an amine-containing group on the head group. In some embodiments, an ionizable lipid is or comprises a compound described herein. In some embodiments, an ionizable lipid is present in a lipid nanoparticle (LNP) preparation from about 30 mole percent to about 70 mole percent, based on total moles of components of the lipid nanoparticle. In some embodiments, an ionizable lipid is present from about 33 mol percent to about 60 mole percent, based on total moles of components of the lipid nanoparticle. In some embodiments, an ionizable lipid is present from about 34 mol percent to about 55 mole percent, based on total moles of components of the lipid nanoparticle. In some embodiments, an ionizable lipid is present from about 33 mol percent to about 51 mole percent, based on total moles of components of the lipid nanoparticle. In some embodiments, an ionizable lipid is present at about 34.7 mole percent, based on total moles of components of the lipid nanoparticle. In some embodiments, an ionizable lipid is present at about 50 mole percent, based on total moles of components of the lipid nanoparticle. [0157] Among other things, in some embodiments, a lipid nanoparticle composition comprises an ionizable lipid. In some embodiments, a lipid nanoparticle preparation comprises an ionizable lipid; a phospholipid; a conjugate-linker lipid; and a cholesterol. In some embodiments, an ionizable lipid is or comprises a structure according to a compound described herein. In some embodiments, an ionizable lipid is present in a LNP preparation from about 30 mole percent to about 70 mole percent, based on total moles of components of the lipid nanoparticle. D. Sterols [0158] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that comprise one or more sterols as described herein. [0159] In some embodiments, a sterol is a cholesterol, or a variant or derivative thereof. In some embodiments, a cholesterol is modified. In some embodiments, a cholesterol is an oxidized cholesterol. In some embodiments, a cholesterol is esterified cholesterol. Unmodified cholesterol can be acted upon by enzymes to form variants that are side-chain or ring oxidized. In some embodiments, a cholesterol can be oxidized on the beta-ring structure or on the hydrocarbon tail structure. In some embodiments, a sterol is a phytosterol. Exemplary sterols that are considered for use in the disclosed lipid nanoparticles include but are not limited to 25-hydroxycholesterol (25-OH), 20α-hydroxycholesterol (20α-OH), 27-hydroxycholesterol, 6-keto-5α- hydroxycholesterol, 7-ketocholesterol, 7β-hydroxycholesterol, 7α-hydroxycholesterol, 7β-25- dihydroxycholesterol, beta-sitosterol, stigmasterol, brassicasterol, campesterol, or combinations thereof. In some embodiments, a side-chain oxidized cholesterol can enhance cargo delivery relative to other cholesterol variants. In some embodiments, a cholesterol is an unmodified cholesterol. [0160] In some embodiments, a LNP composition comprises from about 20 mol percent to about 50 mol percent sterol. In some embodiments, a LNP composition comprises about 38 mol percent sterol. In some embodiments, a LNP composition comprises about 38.5 mol percent sterol. In some embodiments, a LNP composition comprises about 33.8 mol percent cholesterol. E. Conjugate-linker lipids [0161] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that comprise one or more conjugate-linker lipids as described herein. [0162] In some embodiments, a conjugate-linker lipid is or comprises a polyethylene glycol (PEG)-lipid or PEG-modified lipid. In some embodiments, PEG or PEG-modified lipids may be alternately referred to as PEGylated lipids or PEG-lipids. Inclusion of a PEGylating lipid can be used to enhance lipid nanoparticle colloidal stability in vitro and circulation time in vivo. In some embodiments, the PEGylation is reversible in that the PEG moiety is gradually released in blood circulation. Exemplary PEG-lipids include but are not limited to PEG conjugated to saturated or unsaturated alkyl chains having a length of C6-C20. PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides (PEG-CER), PEG-modified dialkylamines, PEG-modified diacylglycerols (PEG-DAG), PEG-modified dialkylglycerols, and mixtures thereof. For example, in some embodiments, a PEG lipid may be PEG-c-DOMG, PEG- DMG, PEG-DLPE, PEG-DMPE, PEG-DPPE, PEG-DSG or a PEG-DSPE lipid. [0163] In some embodiments, a conjugate-linker lipid comprises a polyethylene glycol lipid. In some embodiments, the conjugate-linker lipid comprises DiMystyrlGlycerol (DMG), 1,2- Dipalmitoyl-rac-glycerol, methoxypolyethylene Glycol (DPG-PEG), or 1,2-Distearoyl-rac- glycero-3-methylpolyoxyethylene (DSG – PEG). In some embodiments, a conjugate-linker lipid has an average molecular mass from about 500 Da to about 5000 Da. In some embodiments, a conjugate-linker lipid has an average molecular mass of about 2000 Da. In some embodiments, a LNP composition comprises from about 0 mol percent to about 5 mol percent conjugate-linker lipid. In some embodiments, a LNP composition comprises about 1.5 mol percent conjugate-linker lipid. In some embodiments, a LNP composition comprises about 3 mol percent conjugate-linker lipid. F. Phospholipids [0164] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that comprise one or more phospholipids as described herein. In some embodiments, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that comprise one or more (poly)unsaturated lipids. [0165] In some embodiments, one or more phospholipids may assemble into one or more lipid bilayers. In some embodiments, one or more phospholipids may include a phospholipid moiety. In some embodiments, one or more phospholipids may include one or more fatty acid moieties. In some embodiments, one or more phospholipids may include a phospholipid moiety and one or more fatty acid moieties. In some embodiments, a phospholipid moiety includes but is not limited to phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl serine, phosphatidic acid, 2-lysophosphatidyl choline, and sphingomyelin. In some embodiments, a fatty acid moiety includes but is not limited to lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alphalinolenic acid, erucic acid, phytanic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid. Non-natural species including natural species with modifications and substitutions including branching, oxidation, cyclization, and alkynes are also contemplated. For example, a phospholipid may be functionalized with or cross-linked to one or more alkynes (e.g., an alkenyl group in which one or more double bonds is replaced with a triple bond). Under appropriate reaction conditions, an alkyne group may undergo a copper-catalyzed cycloaddition upon exposure to an azide. Such reactions may be useful in functionalizing a lipid bilayer of a nanoparticle composition to facilitate membrane permeation or cellular recognition or in conjugating a nanoparticle composition to a useful component such as a targeting or imaging moiety (e.g., a dye). [0166] Exemplary phospholipids include but are not limited to 1,2-distearoyl-snglycero-3- phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2- dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycerophosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycerophosphocholine (DUPC), l-palmitoyl-2- oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoy l-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl snglycero-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-diphytanoyl-sn-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), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearoyl-phosphatidyl-ethanolamine (DSPE), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidy ethanolamine (SOPE), 1-stearoyl-2 oleoylphosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoyl phosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), or combinations thereof. In some embodiments, a phospholipid is DSPC. In some embodiments, a phospholipid is DMPC. [0167] In some embodiments, the phospholipid comprises 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine-N-(succinyl) (succinyl PE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-dipalmitoyl- sn-glycero-3-phosphoethanolamine-N-(succinyl) (succinyl-DPPE), 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2- dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), or a combination thereof. G. Diameter [0168] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that have an average hydrodynamic diameter from about 30 to about 220 nm. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have an average hydrodynamic diameter that is about 30 nm, 35 nm,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, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have an average hydrodynamic diameter from between 50 nm to 200 nm. [0169] In some embodiments, lipid nanoparticles described herein have an average hydrodynamic diameter from about 30 to about 220 nm. In some embodiments, lipid nanoparticles described herein have an average hydrodynamic diameter that is about 30 nm, 35 nm,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, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, lipid nanoparticles described herein have an average hydrodynamic diameter from between 50 nm to 200 nm. H. Polydispersity [0170] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that have a polydispersity index (PDI) of about 0.01 to about 0.3. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have a PDI that is about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have a PDI from about 0.05 to about 0.2, about 0.06 to about 0.1, or about 0.07 to about 0.09. [0171] In some embodiments, lipid nanoparticles described herein have a PDI from about 0.01 to about 0.3. In some embodiments, lipid nanoparticles described herein have a PDI that is about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, lipid nanoparticles described herein have a PDI from about 0.05 to about 0.2, about 0.06 to about 0.1, or about 0.07 to about 0.09. I. Encapsulation efficiency [0172] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials, wherein encapsulation efficiency of provided compositions, preparations, nanoparticles, and/or nanomaterials is from about 80% to about 100%. In some embodiments, encapsulation efficiency of compositions, preparations, nanoparticles, and/or nanomaterials described herein is about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 100%, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, encapsulation efficiency of compositions, preparations, nanoparticles, and/or nanomaterials described herein is from about 90% to about 100%, about 95% to about 100%, about 95% to about 98%, or about 95.5% to about 97.5%. In some embodiments, encapsulation efficiency of compositions, preparations, nanoparticles, and/or nanomaterials described herein is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. [0173] In some embodiments, encapsulation efficiency of lipid nanoparticles described herein is from about 80% to about 100%. In some embodiments, encapsulation efficiency of lipid nanoparticles described herein is about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 100%, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, encapsulation efficiency of lipid nanoparticles described herein is from about 90% to about 100%, about 95% to about 100%, about 95% to about 98%, or about 95.5% to about 97.5%. In some embodiments, encapsulation efficiency of lipid nanoparticles described herein is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. J. pKa [0174] Among other things, the present disclosure describes compositions, preparations, nanoparticles, and/or nanomaterials that have a pKa from about 5 to about 9. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have a pKa that is about 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, or any range having endpoints defined by any two of the aforementioned values. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein have a pKa that is about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, or any range having endpoints defined by any two of the aforementioned values. [0175] In some embodiments, lipid nanoparticles described herein have a pKa from about 5 to about 9. In some embodiments, lipid nanoparticles described herein have a pKa that is about 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, or any range having endpoints defined by any two of the aforementioned values. In some embodiments, lipid nanoparticles described herein have a pKa that is about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, or any range having endpoints defined by any two of the aforementioned values. II. Exemplary LNP Preparations [0176] The present invention provides for compositions, preparations, nanoparticles, and/or nanomaterials that comprise lipid nanoparticles. In some embodiments, a lipid nanoparticle preparation comprises about 30 mole percent to about 70 mole percent ionizable lipid, about 5 mole percent to about 25 mole percent phospholipid, about 25 mole percent to about 45 mole percent cholesterol, and about 0 mole percent to about 5 mole percent conjugate-linker lipid. [0177] In some embodiments, a lipid nanoparticle preparation comprises about 45 mole percent ionizable lipid, about 9 mole percent phospholipid, about 44 mole percent cholesterol, and about 2 mole percent conjugate-linker lipid. In some embodiments, a lipid nanoparticle preparation comprises about 50 mole percent ionizable lipid, about 9 mole percent phospholipid, about 38 mole percent cholesterol, and about 3 mole percent conjugate-linker lipid. [0178] In some embodiments, a lipid nanoparticle preparation comprises about 40 mole percent to about 60 mole percent ionizable lipid of any provided compound, about 5 mole percent to about 15 mole percent 1-2-distearoyl-sn-glycero-3-phosphocholine, about 1 mole percent to about 5 mole percent C14PEG2000, and about 30 mole percent to about 47 mole percent cholesterol, based on the total moles of these four ingredients. [0179] In some embodiments, a lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA from about 2:1 and 50:1. In some embodiments, a LNP preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1. In some embodiments, a lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 11.7:1 and 19:1. [0180] In some embodiments, a lipid nanoparticle preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):siRNA from about 2:1 and 50:1. In some embodiments, a LNP preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1. In some embodiments, a lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 11.7:1 and 19:1. [0181] In some embodiments, an LNP preparation comprises one or more nucleic acids such as RNAs. In some embodiments, one or more nucleic acids (e.g., RNAs), lipids, and amounts thereof in an LNP preparation may be selected to provide a specific N/P ratio. The N/P ratio can be selected from about 1 to about 30. The N/P ratio can be selected from about 2 to about 12. In some embodiments, the N/P ratio is from about 0.1 to about 50. In some embodiments, the N/P ratio is from about 2 to about 8. In some embodiments, the N/P ratio is from about 2 to about 15, from about 2 to about 10, from about 2 to about 8, from about 2 to about 6, from about 3 to about 15, from about 3 to about 10, from about 3 to about 8, from about 3 to about 6, from about 4 to about 15, from about 4 to about 10, from about 4 to about 8, from about 4 to about 6, or from about 5 to about 7. In some embodiments, the N/P ratio is about 2, about 2.5, about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, about 9, or about 10. In some embodiments, the N/P ratio is from about 4 to about 6. In some embodiments, the N/P ratio is about 4, about 4.5, about 5, about 5.5, or about 6. [0182] As used herein, the “N/P ratio” is the molar ratio of ionizable (e.g., in the physiological pH range) nitrogen atoms in a lipid (or lipids) to phosphate groups in a nucleic acid molecular entity (or nucleic acid molecular entities), e.g., in a nanoparticle composition comprising a lipid component and an RNA. Ionizable nitrogen atoms can include, for example, nitrogen atoms that can be protonated at about pH 1, about pH 2, about pH 3, about pH 4, about pH 4.5, about pH 5, about pH 5.5, about pH 6, about pH 6.5, about pH 7, about pH 7.5, or about pH 8 or higher. The physiological pH range can include, for example, the pH range of different cellular compartments (such as organs, tissues, and cells) and bodily fluids (such as blood, CSF, gastric juice, milk, bile, saliva, tears, and urine). In certain specific embodiments, the physiological pH range refers to the pH range of blood in a mammal, for example, from about 7.35 to about 7.45. Similarly, for phosphate charge neutralizers that have one or more ionizable nitrogen atoms, the N/P ratio can refer to a molar ratio of ionizable nitrogen atoms in the phosphate charge neutralizer to the phosphate groups in a nucleic acid. In some embodiments, ionizable nitrogen atoms refer to those nitrogen atoms that are ionizable within a pH range between 5 and 14. III. Pharmaceutical compositions [0183] The present invention provides for compositions, preparations, nanoparticles, and/or nanomaterials that comprise pharmaceutical compositions. Among other things, in some embodiments, pharmaceutical compositions comprise lipid nanoparticles and lipid nanoparticle preparations described herein. For example, in some embodiments, lipid nanoparticles and lipid nanoparticle preparations described herein can be formulated in whole or in part as pharmaceutical compositions. [0184] In some embodiments, pharmaceutical compositions may include one or more nanoparticle compositions described herein. For example, a pharmaceutical composition may comprise one or more nanoparticle compositions including one or more different therapeutic and/or prophylactics including but not limited to one or more nucleic acids of different types or encode different agents. In some embodiments, a pharmaceutical composition comprises one or more pharmaceutically acceptable excipients or accessory ingredients including but not limited to a pharmaceutically acceptable carrier. [0185] A pharmaceutical composition may be administered to a subject. In some embodiments, a pharmaceutical composition is administered as described herein. In some in vivo approaches, the nanoparticle compositions disclosed herein are administered to a subject in a therapeutically effective amount as described herein. [0186] In some embodiments, the ordinary skilled worker, considering the therapeutic context, age, and general health of the recipient, will be able to devise an appropriate dosage level and dosing regimen using the pharmaceutical compositions described herein for treatment of various conditions in various patients. For example, in some embodiments, a selected dosage depends upon the desired therapeutic effect, on the route of administration, and on the duration of the treatment desired. In some embodiments, generally dosage levels of about 0.001 mg to about 5 mg of nucleic acid per kg of body weight are administered each dosage to mammals. More specifically, in some embodiments, a preferential dose for nucleic acids within the disclosed nanoparticles is about 0.1 mg / kg to about 1.0 mg/kg. For the disclosed nanoparticles, generally dosage levels of about 0.2 mg to about 100 mg of four components (ionizable lipid, cholesterol, conjugate-linker conjugate, and phospholipid) / kg of body weight are administered to mammals. More specifically, in some embodiments, a preferential dose of the disclosed nanoparticles is about 0.5 mg / kg to about 5 mg / kg of the four components / kg of body weight. [0187] In some embodiments, a pharmaceutical composition described herein is administered locally, for example by injection directly into a site to be treated. Typically, the injection causes an increased localized concentration of the composition which is greater than that which can be achieved by systemic administration. In some embodiments, a pharmaceutical composition described herein can be combined with a matrix as described herein to assist in creating an increased localized concentration of the polypeptide compositions by reducing the passive diffusion of the polypeptides out of the site to be treated. A. Preparations for parenteral administration [0188] In some embodiments, the compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein, including those containing lipid nanoparticles, are administered in an aqueous solution, by parenteral injection. In some embodiments, a preparation may also be in the form of a suspension or emulsion. In general, pharmaceutical compositions are provided including effective amounts of a lipid nanoparticle, and optionally include pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers. Such compositions optionally include one or more for the following: diluents, sterile water, buffered saline of various buffer content (e.g., Tris-HCl, acetate, phosphate), pH and ionic strength; and additives such as detergents and solubilizing agents (e.g., TWEEN 20 (polysorbate-20), TWEEN 80 (polysorbate- 80)), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), and preservatives (e.g., Thimersol, benzyl alcohol) and bulking substances (e.g., lactose, mannitol). Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate. Formulations may be lyophilized and redissolved/resuspended immediately before use. A formulation may be sterilized by, for example, filtration through a bacteria retaining filter, by incorporating sterilizing agents into the compositions, by irradiating the compositions, or by heating the compositions. B. Controlled delivery polymeric matrices [0189] In some embodiments, the compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein can also be administered in controlled release formulations. In some embodiments, controlled release polymeric devices can be made for long term release systemically following implantation of a polymeric device (such as a rod, cylinder, film, disk) or injection (such as microparticles). In some embodiments, a matrix can be in the form of microparticles such as microspheres. In some embodiments, an agent is dispersed within a solid polymeric matrix or microcapsules. In some embodiments, a core is of a different material than a polymeric shell of any of the described compositions, preparations, nanoparticles, and/or nanomaterials. In some embodiments, a peptide is dispersed or suspended in a core, which may be liquid or solid in nature, of any of the described compositions, preparations, nanoparticles, and/or nanomaterials. Unless specifically defined herein, microparticles, microspheres, and microcapsules are used interchangeably. In some embodiments, a polymer may be cast as a thin slab or film, ranging from nanometers to four centimeters, a powder produced by grinding or other standard techniques, or even a gel such as a hydrogel. [0190] In some embodiments, non-biodegradable matrices are used for delivery of the described compositions, preparations, nanoparticles, and/or nanomaterials. In some embodiments, biodegradable matrices are used for delivery of the described compositions, preparations, nanoparticles, and/or nanomaterials. In some embodiments, biodegradable matrices are preferred. In some embodiments, biodegradable matrices comprise natural or synthetic polymers. In some embodiments, synthetic polymers are preferred due to the better characterization of degradation and release profiles. In some embodiments, a polymer is selected based on the period over which release is desired. In some embodiments, linear release may be most useful, although in others a pulse release or “bulk release” may provide more effective results. In some embodiments, a polymer may be in the form of a hydrogel (typically in absorbing up to about 90% by weight of water), and can optionally be crosslinked with multivalent ions or polymers. [0191] The matrices can be formed by solvent evaporation, spray drying, solvent extraction and other methods known to those skilled in the art. Bioerodible microspheres can be prepared using any of the methods developed for making microspheres for drug delivery, for example, as described by Mathiowitz and Langer, J. Controlled Release, 5:13-22 (1987); Mathiowitz, et al., Reactive Polymers, 6:275-283 (1987); and Mathiowitz, et al., J. Appl. Polymer Sci., 35:755-774 (1988), the disclosure of which is hereby incorporated by reference in its entirety herein. [0192] In some embodiments, the described compositions, preparations, nanoparticles, and/or nanomaterials can be formulated for local release to treat the area of implantation or injection – which will typically deliver a dosage that is much less than the dosage for treatment of an entire body – or systemic delivery. These can be implanted or injected subcutaneously, into the muscle, fat, or swallowed. C. Cargo [0193] Among other things, the present invention provides for compositions, preparations, nanoparticles, and/or nanomaterials that comprise cargo as described herein. In some embodiments, the compositions, preparations, nanoparticles, and/or nanomaterials include a therapeutic or prophylactic agent for delivery to a subject. In some embodiments, a therapeutic or prophylactic agent is encapsulated by a lipid nanoparticle. In some embodiments, a lipid nanoparticle is loaded with one or more nucleic acids. D. Therapeutic and/or prophylactic agents [0194] Cargo delivered via a LNP composition may be a biologically active agent. In some embodiments, the cargo is or comprises one or more biologically active agents, such as mRNA, guide RNA (gRNA), nucleic acid, RNA-guided DNA-binding agent, expression vector, template nucleic acid, antibody (e.g. , monoclonal, chimeric, humanized, nanobody, and fragments thereof etc.), cholesterol, hormone, peptide, protein, chemotherapeutic and other types of antineoplastic agent, low molecular weight drug, vitamin, co-factor, nucleoside, nucleotide, oligonucleotide, enzymatic nucleic acid, antisense nucleic acid, triplex forming oligonucleotide, antisense DNA or RNA composition, chimeric DNA:RNA composition, allozyme, aptamer, ribozyme, decoys and analogs thereof, plasmid and other types of vectors, and small nucleic acid molecule, RNAi agent, short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA) and “self-replicating RNA” (encoding a replicase enzyme activity and capable of directing its own replication or amplification in vivo) molecules, peptide nucleic acid (PNA), a locked nucleic acid ribonucleotide (LNA), morpholino nucleotide, threose nucleic acid (TNA), glycol nucleic acid (GNA), sisiRNA (small internally segmented interfering RNA), and iRNA (asymmetrical interfering RNA). The above list of biologically active agents is exemplary only, and is not intended to be limiting. Such compounds may be purified or partially purified, and may be naturally occurring or synthetic, and may be chemically modified. [0195] Cargo delivered via a LNP composition may be an RNA, such as an mRNA molecule encoding a protein of interest. For example, in some embodiments, an mRNA for expressing a protein such as green fluorescent protein (GFP), an RNA-guided DNA-binding agent, or a Cas nuclease is described herein. LNP compositions that include a Cas nuclease mRNA, for example a Class 2 Cas nuclease mRNA that allows for expression in a cell of a Class 2 Cas nuclease such as a Cas9 or Cpfl protein are provided. Further, cargo may contain one or more guide RNAs or nucleic acids encoding guide RNAs. A template nucleic acid, e.g., for repair or recombination, may also be included in the composition or a template nucleic acid may be used in the methods described herein. In some embodiments, cargo comprises an mRNA that encodes a Streptococcus pyogenes Cas9, optionally and an S. pyogenes gRNA. In some embodiments, cargo comprises an mRNA that encodes a Neisseria meningitidis Cas9, optionally and an nme gRNA. [0196] “mRNA” refers to a polynucleotide and comprises an open reading frame that can be translated into a polypeptide (i.e., can serve as a substrate for translation by a ribosome and amino- acylated tRNAs). mRNA can comprise a phosphate-sugar backbone including ribose residues or analogs thereof, e.g., 2’-methoxy ribose residues. In some embodiments, the sugars of an mRNA phosphate-sugar backbone consist essentially of ribose residues, 2’-methoxy ribose residues, or a combination thereof. In general, mRNAs do not contain a substantial quantity of thymidine residues (e.g., 0 residues or fewer than 30, 20, 10, 5, 4, 3, or 2 thymidine residues; or less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, or 0.1% thymidine content). An mRNA can contain modified uridines at some or all of its uridine positions. E. CRISPR/Cas Cargo [0197] In some embodiments, the disclosed compositions, preparations, nanoparticles, and/or nanomaterials comprise an mRNA encoding an RNA-guided DNA-binding agent, such as a Cas nuclease. In particular embodiments, the disclosed compositions, preparations, nanoparticles, and/or nanomaterials comprise an mRNA encoding a Class 2 Cas nuclease, such as S. pyogenes Cas9. [0198] As used herein, an “RNA-guided DNA binding agent” means a polypeptide or complex of polypeptides having RNA and DNA binding activity, or a DNA-binding subunit of such a complex, wherein the DNA binding activity is sequence-specific and depends on the sequence of the RNA. Exemplary RNA-guided DNA binding agents include Cas cleavases/nickases and inactivated forms thereof (“dCas DNA binding agents”). “Cas nuclease”, as used herein, encompasses Cas cleavases, Cas nickases, and dCas DNA binding agents. Cas cleavases/nickases and dCas DNA binding agents include a Csm or Cmr complex of a type III CRISPR system, the CaslO, Csml, or Cmr2 subunit thereof, a Cascade complex of a type I CRISPR system, the Cas3 subunit thereof, and Class 2 Cas nucleases. As used herein, a“Class 2 Cas nuclease” is a single chain polypeptide with RNA-guided DNA binding activity. Class 2 Cas nucleases include Class 2 Cas cleavases/nickases (e.g., H840A, D10A, or N863 A variants), which further have RNA-guided DNA cleavases or nickase activity, and Class 2 dCas DNA binding agents, in which cleavase/nickase activity is inactivated. Class 2 Cas nucleases include, for example, Cas9, Cpfl, C2cl, C2c2, C2c3, HF Cas9 (e.g., N497A, R661A, Q695A, Q926A variants), HypaCas9 (e.g., N692A, M694A, Q695A, H698A variants), eSPCas9(1.0) (e.g., K810A, K1003A, R1060A variants), and eSPCas9(1.1) (e.g., K848A, K1003A, R1060A variants) proteins and modifications thereof. Cpfl protein, Zetsche et al., Cell, 163: 1-13 (2015), is homologous to Cas9, and contains a RuvC-like nuclease domain. Cpfl sequences of Zetsche are incorporated by reference in their entirety herein. See, e.g., Zetsche, Tables Sl and S3. See, e.g, Makarova et al., Nat Rev Microbiol, 13(11): 722-36 (2015); Shmakov et al., Molecular Cell, 60:385-397 (2015), the contents of which are hereby incorporated in its entirety herein. [0199] As used herein, “ribonucleoprotein” (RNP) or “RNP complex” refers to a guide RNA together with an RNA-guided DNA binding agent, such as a Cas nuclease, e.g., a Cas cleavase, Cas nickase, or dCas DNA binding agent (e.g., Cas9). In some embodiments, the guide RNA guides the RNA-guided DNA binding agent such as Cas9 to a target sequence, and the guide RNA hybridizes with and the agent binds to the target sequence; in cases where the agent is a cleavase or nickase, binding can be followed by cleaving or nicking. [0200] In some embodiments, cargo for a LNP composition includes at least one guide RNA comprising guide sequences that direct an RNA-guided DNA binding agent, which can be a nuclease (e.g., a Cas nuclease such as Cas9), to a target DNA. gRNA may guide the Cas nuclease or Class 2 Cas nuclease to a target sequence on a target nucleic acid molecule. In some embodiments, a gRNA binds with and provides specificity of cleavage by a Class 2 Cas nuclease. In some embodiments, a gRNA and a Cas nuclease may form a ribonucleoprotein (RNP), e.g., a CRISPR/Cas complex such as a CRISPR/Cas9 complex. In some embodiments, a CRISPR/Cas complex may be a Type-II CRISPR/Cas9 complex. In some embodiments, a CRISPR/Cas complex may be a Type-V CRISPR/Cas complex, such as a Cpfl/guide RNA complex. Cas nucleases and cognate gRNAs may be paired. A gRNA scaffold structures that pair with each Class 2 Cas nuclease vary with the specific CRISPR/Cas system. [0201] “Guide RNA”, “gRNA”, and simply “guide” are used herein interchangeably to refer to either a crRNA (also known as CRISPR RNA), or the combination of a crRNA and a trRNA (also known as tracrRNA). Guide RNAs can include modified RNAs as described herein. The crRNA and trRNA may be associated as a single RNA molecule (single guide RNA, sgRNA) or in two separate RNA molecules (dual guide RNA, dgRNA). “Guide RNA” or “gRNA” refers to each type. trRNA may be a naturally-occurring sequence, or a trRNA sequence with modifications or variations compared to naturally-occurring sequences. [0202] As used herein, a “guide sequence” refers to a sequence within a guide RNA that is complementary to a target sequence and functions to direct a guide RNA to a target sequence for binding or modification (e.g., cleavage) by an RNA-guided DNA binding agent. A “guide sequence” may also be referred to as a “targeting sequence,” or a “spacer sequence.” A guide sequence can be 20 base pairs in length, e.g., in the case of Streptococcus pyogenes (i.e., Spy Cas9) and related Cas9 homologs/orthologs. Shorter or longer sequences can also be used as guides, e.g., 15-, 16-, 17-, 18-, 19-, 21-, 22-, 23-, 24-, or 25-nucleotides in length. In some embodiments, a target sequence is in a gene or on a chromosome, for example, and is complementary to a guide sequence. In some embodiments, a degree of complementarity or identity between a guide sequence and its corresponding target sequence may be about or at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, a guide sequence and the target region may be 100% complementary or identical over a region of at least 15, 16, 17, 18, 19, or 20 contiguous nucleotides. In other embodiments, a guide sequence and a target region may contain at least one mismatch. For example, a guide sequence and a target sequence may contain 1, 2, 3, or 4 mismatches, where the total length of the target sequence is at least 17, 18, 19, 20 or more base pairs. In some embodiments, a guide sequence and a target region may contain 1-4 mismatches where a guide sequence comprises at least 17, 18, 19, 20 or more nucleotides. In some embodiments, a guide sequence and the target region may contain 1, 2, 3, or 4 mismatches where the guide sequence comprises 20 nucleotides. [0203] Target sequences for RNA-guided DNA binding proteins such as Cas proteins include both the positive and negative strands of genomic DNA (i.e., the sequence given and the sequence’s reverse compliment), as a nucleic acid substrate for a Cas protein is a double stranded nucleic acid. Accordingly, where a guide sequence is said to be “complementary to a target sequence”, it is to be understood that the guide sequence may direct a guide RNA to bind to the reverse complement of a target sequence. Thus, in some embodiments, where the guide sequence binds the reverse complement of a target sequence, the guide sequence is identical to certain nucleotides of the target sequence (e.g., the target sequence not including the PAM) except for the substitution of U for T in the guide sequence. [0204] The length of the targeting sequence may depend on the CRISPR/Cas system and components used. For example, different Class 2 Cas nucleases from different bacterial species have varying optimal targeting sequence lengths. Accordingly, the targeting sequence may comprise 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, 35, 40, 45, 50, or more than 50 nucleotides in length. In some embodiments, the targeting sequence length is 0, 1, 2, 3, 4, or 5 nucleotides longer or shorter than the guide sequence of a naturally-occurring nucleotide sequence. [0205] In certain embodiments, a Cas nuclease and gRNA scaffold will be derived from the same CRISPR/Cas system. In some embodiments, a targeting sequence may comprise or consist of 18- 24 nucleotides. In some embodiments, a targeting sequence may comprise or consist of 19-21 nucleotides. In some embodiments, the targeting sequence may comprise or consist of 20 nucleotides. [0206] In some embodiments, a sgRNA is a “Cas9 sgRNA” capable of mediating RNA-guided DNA cleavage by a Cas9 protein. In some embodiments, a sgRNA is a “Cpfl sgRNA” capable of mediating RNA-guided DNA cleavage by a Cpfl protein. In some embodiments, a gRNA comprises a crRNA and tracr RNA sufficient for forming an active complex with a Cas9 protein and mediating RNA-guided DNA cleavage. In some embodiments, a gRNA comprises a crRNA sufficient for forming an active complex with a Cpfl protein and mediating RNA-guided DNA cleavage. See Zetsche 2015. [0207] Certain embodiments of the invention also provide nucleic acids, e.g., expression cassettes, encoding the gRNA described herein. A “guide RNA nucleic acid” is used herein to refer to a guide RNA (e.g. an sgRNA or a dgRNA) and a guide RNA expression cassette, which is a nucleic acid that encodes one or more guide RNAs. [0208] Certain embodiments of the present disclosure also provide delivery of adenine base editors (“ABEs”) using the LNPs compositions, preparations, nanoparticles, and/or nanomaterials described herein. ABEs and methods of their use are described, e.g. in U.S. Patent No.10,113,163 and U.S. Patent Publication No. 2021/0130805, the contents of each of which are hereby incorporated by reference in their entireties. [0209] Certain embodiments of the present disclosure also provide delivery of cytosine base editors (“CBEs”) using the LNPs compositions, preparations, nanoparticles, and/or nanomaterials described herein. ABEs and methods of their use are described, e.g. in U.S. Patent Nos.10,167,457 and 9,840,699, the contents of each of which are hereby incorporated by reference in their entireties. [0210] The term “base editor (BE),” or “nucleobase editor (NBE)” refers to an agent comprising a polypeptide that is capable of making a modification to a base (e.g., A, T, C, G, or U) within a nucleic acid sequence (e.g., DNA or RNA). In some embodiments, the base editor is capable of deaminating a base within a nucleic acid. In some embodiments, the base editor is capable of deaminating a base within a DNA molecule. In some embodiments, the base editor is capable of deaminating an adenine (A) in DNA. . In some embodiments, the deaminase is a cytosine deaminase or a cytidine deaminase. In some embodiments, the base editor is a fusion protein comprising a nucleic acid programmable DNA binding protein (napDNAbp) fused to an adenosine deaminase. In some embodiments, the base editor is a Cas9 protein fused to an adenosine deaminase. In some embodiments, the base editor is a Cas9 nickase (nCas9) fused to an adenosine deaminase. In some embodiments, the base editor is a nuclease-inactive Cas9 (dCas9) fused to an adenosine deaminase. In some embodiments, the base editor is fused to an inhibitor of base excision repair, for example, a UGI domain, or a dISN domain. In some embodiments, the fusion protein comprises a Cas9 nickase fused to a deaminase and an inhibitor of base excision repair, such as a UGI or dISN domain. The term “nucleic acid programmable DNA binding protein” or “napDNAbp” refers to a protein that associates with a nucleic acid (e.g., DNA or RNA), such as a guide nuclic acid, that guides the napDNAbp to a specific nucleic acid sequence. For example, a Cas9 protein can associate with a guide RNA that guides the Cas9 protein to a specific DNA sequence that has complementary to the guide RNA. In some embodiments, the napDNAbp is a class 2 microbial CRISPR-Cas effector. In some embodiments, the napDNAbp is a Cas9 domain, for example a nuclease active Cas9, a Cas9 nickase (nCas9), or a nuclease inactive Cas9 (dCas9). Examples of nucleic acid programmable DNA binding proteins include, without limitation, Cas9 (e.g., dCas9 and nCas9), CasX, CasY, Cpf1, C2c1, C2c2, C2C3, and Argonaute. It should be appreciated, however, that nucleic acid programmable DNA binding proteins also include nucleic acid programmable proteins that bind RNA. For example, the napDNAbp may be associated with a nucleic acid that guides the napDNAbp to an RNA. Other nucleic acid programmable DNA binding proteins are also within the scope of this disclosure, though they may not be specifically listed in this disclosure. F. Modified RNAs [0211] In certain embodiments, the disclosed compositions, preparations, nanoparticles, and/or nanomaterials comprise modified nucleic acids, including modified RNAs. [0212] Modified nucleosides or nucleotides can be present in an RNA, for example a gRNA or mRNA. A gRNA or mRNA comprising one or more modified nucleosides or nucleotides, for example, is called a “modified” RNA to describe the presence of one or more non-naturally and/or naturally occurring components or configurations that are used instead of or in addition to the canonical A, G, C, and U residues. In some embodiments, a modified RNA is synthesized with a non-canonical nucleoside or nucleotide, here called “modified.” [0213] Modified nucleosides and nucleotides can include one or more of: (i) alteration, e.g., replacement, of one or both of the non-linking phosphate oxygens and/or of one or more of the linking phosphate oxygens in the phosphodiester backbone linkage (an exemplary backbone modification); (ii) alteration, e.g., replacement, of a constituent of the ribose sugar, e.g. , of the 2' hydroxyl on the ribose sugar (an exemplary sugar modification); (iii) wholesale replacement of the phosphate moiety with “dephospho” linkers (an exemplary backbone modification); (iv) modification or replacement of a naturally occurring nucleobase, including with a non-canonical nucleobase (an exemplary base modification); (v) replacement or modification of the ribose- phosphate backbone (an exemplary backbone modification); (vi) modification of the 3' end or 5' end of the oligonucleotide, e.g. , removal, modification or replacement of a terminal phosphate group or conjugation of a moiety, cap or linker (such 3' or 5' cap modifications may comprise a sugar and/or backbone modification); and (vii) modification or replacement of the sugar (an exemplary sugar modification). Certain embodiments comprise a 5' end modification to an mRNA, gRNA, or nucleic acid. Certain embodiments comprise a 3' end modification to an mRNA, gRNA, or nucleic acid. A modified RNA can contain 5' end and 3' end modifications. A modified RNA can contain one or more modified residues at non-terminal locations. In certain embodiments, a gRNA includes at least one modified residue. In certain embodiments, an mRNA includes at least one modified residue. [0214] Unmodified nucleic acids can be prone to degradation by, e.g., intracellular nucleases or those found in serum. For example, nucleases can hydrolyze nucleic acid phosphodiester bonds. Accordingly, in one aspect the RNAs (e.g. mRNAs, gRNAs) described herein can contain one or more modified nucleosides or nucleotides, e.g., to introduce stability toward intracellular or serum- based nucleases. In some embodiments, the modified gRNA molecules described herein can exhibit a reduced innate immune response when introduced into a population of cells, both in vivo and ex vivo. The term “innate immune response” includes a cellular response to exogenous nucleic acids, including single stranded nucleic acids, which involves the induction of cytokine expression and release, particularly the interferons, and cell death. [0215] Accordingly, in some embodiments, RNA or nucleic acids in the disclosed the disclosed compositions, preparations, nanoparticles, and/or nanomaterials comprise at least one modification which confers increased or enhanced stability to the nucleic acid, including, for example, improved resistance to nuclease digestion in vivo. As used herein, the terms “modification” and “modified” as such terms relate to the nucleic acids provided herein, include at least one alteration which preferably enhances stability and renders the RNA or nucleic acid more stable (e.g., resistant to nuclease digestion) than the wild-type or naturally occurring version of the RNA or nucleic acid. As used herein, the terms “stable” and “stability” as such terms relate to the nucleic acids of the present invention, and particularly with respect to the RNA, refer to increased or enhanced resistance to degradation by, for example nucleases (i.e., endonucleases or exonucleases) which are normally capable of degrading such RNA. Increased stability can include, for example, less sensitivity to hydrolysis or other destruction by endogenous enzymes (e.g., endonucleases or exonucleases) or conditions within the target cell or tissue, thereby increasing or enhancing the residence of such RNA in the target cell, tissue, subject and/or cytoplasm. The stabilized RNA molecules provided herein demonstrate longer half-lives relative to their naturally occurring, unmodified counterparts (e.g. the wild-type version of the mRNA). Also contemplated by the terms “modification” and “modified” as such terms related to the mRNA of the LNP compositions disclosed herein are alterations which improve or enhance translation of mRNA nucleic acids, including for example, the inclusion of sequences which function in the initiation of protein translation (e.g., the Kozac consensus sequence). (Kozak, M., Nucleic Acids Res 15 (20): 8125- 48 (1987), the contents of which are hereby incorporated by reference herein in its entirety). [0216] In some embodiments, an RNA or nucleic acid of the disclosed compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein have undergone a chemical or biological modification to render it more stable. Exemplary modifications to an RNA include the depletion of a base (e.g., by deletion or by the substitution of one nucleotide for another) or modification of a base, for example, the chemical modification of a base. The phrase “chemical modifications” as used herein, includes modifications which introduce chemistries which differ from those seen in naturally occurring RNA, for example, covalent modifications such as the introduction of modified nucleotides, (e.g., nucleotide analogs, or the inclusion of pendant groups which are not naturally found in such RNA molecules). [0217] In some embodiments of a backbone modification, the phosphate group of a modified residue can be modified by replacing one or more of the oxygens with a different substituent. Further, the modified residue, e.g., modified residue present in a modified nucleic acid, can include the wholesale replacement of an unmodified phosphate moiety with a modified phosphate group as described herein. In some embodiments, the backbone modification of the phosphate backbone can include alterations that result in either an uncharged linker or a charged linker with unsymmetrical charge distribution. Examples of modified phosphate groups include, phosphorothioate, phosphoroselenates, borano phosphates, borano phosphate esters, hydrogen phosphonates, phosphoroamidates, alkyl or aryl phosphonates and phosphotriesters. The phosphorous atom in an unmodified phosphate group is achiral. However, replacement of one of the non-bridging oxygens with one of the above atoms or groups of atoms can render the phosphorous atom chiral. The stereogenic phosphorous atom can possess either the “R” configuration (herein Rp) or the “S” configuration (herein Sp). The backbone can also be modified by replacement of a bridging oxygen, (i.e., the oxygen that links the phosphate to the nucleoside), with nitrogen (bridged phosphoroamidates), sulfur (bridged phosphorothioates) and carbon (bridged methylenephosphonates). The replacement can occur at either linking oxygen or at both of the linking oxygens. The phosphate group can be replaced by non-phosphorus containing connectors in certain backbone modifications. In some embodiments, the charged phosphate group can be replaced by a neutral moiety. Examples of moieties which can replace the phosphate group can include, without limitation, e.g., methyl phosphonate, hydroxylamino, siloxane, carbonate, carboxymethyl, carbamate, amide, thioether, ethylene oxide linker, sulfonate, sulfonamide, thioformacetal, formacetal, oxime, methyleneimino, methylenemethylimino, methylenehydrazo, methylenedimethylhydrazo and methyleneoxymethylimino. G. mRNA [0218] In some embodiments, the disclosed compositions, preparations, nanoparticles, and/or nanomaterials comprise an mRNA comprising an open reading frame (ORF) encoding an RNA- guided DNA binding agent, such as a Cas nuclease, or Class 2 Cas nuclease as described herein. In some embodiments, an mRNA comprising an ORF encoding an RNA-guided DNA binding agent, such as a Cas nuclease or Class 2 Cas nuclease, is provided, used, or administered. An mRNA may comprise one or more of a 5' cap, a 5' untranslated region (UTR), a 3' UTRs, and a polyadenine tail. The mRNA may comprise a modified open reading frame, for example to encode a nuclear localization sequence or to use alternate codons to encode the protein. [0219] mRNA in the disclosed compositions, preparations, nanoparticles, and/or nanomaterials may encode, for example, a secreted hormone, enzyme, receptor, polypeptide, peptide or other protein of interest that is normally secreted. In one embodiment of the invention, the mRNA may optionally have chemical or biological modifications which, for example, improve the stability and/or half-life of such mRNA or which improve or otherwise facilitate protein production. [0220] In addition, suitable modifications include alterations in one or more nucleotides of a codon such that the codon encodes the same amino acid but is more stable than the codon found in the wild-type version of the mRNA. For example, an inverse relationship between the stability of RNA and a higher number cyti dines (C's) and/or uridines (U's) residues has been demonstrated, and RNA devoid of C and U residues have been found to be stable to most RNases (Heidenreich, et al. J Biol Chem 269, 2131-8 (1994), the disclosure of which is hereby incorporated by reference herein in its entirety). In some embodiments, the number of C and/or U residues in an mRNA sequence is reduced. In another embodiment, the number of C and/or U residues is reduced by substitution of one codon encoding a particular amino acid for another codon encoding the same or a related amino acid. Contemplated modifications to the mRNA nucleic acids of the present invention also include the incorporation of pseudouridines. The incorporation of pseudouridines into the mRNA nucleic acids of the present invention may enhance stability and translational capacity, as well as diminishing immunogenicity in vivo. See, e.g., Kariko, K., et al., Molecular Therapy 16 (11): 1833-1840 (2008), the contents of which is hereby incorporated by reference herein in its entirety. Substitutions and modifications to the mRNA of the present invention may be performed by methods readily known to one or ordinary skill in the art. [0221] The constraints on reducing the number of C and U residues in a sequence will likely be greater within the coding region of an mRNA, compared to an untranslated region, (i.e., it will likely not be possible to eliminate all of the C and U residues present in the message while still retaining the ability of the message to encode the desired amino acid sequence). The degeneracy of the genetic code, however presents an opportunity to allow the number of C and/or U residues that are present in the sequence to be reduced, while maintaining the same coding capacity (i.e., depending on which amino acid is encoded by a codon, several different possibilities for modification of RNA sequences may be possible). [0222] The term modification also includes, for example, the incorporation of non-nucleotide linkages or modified nucleotides into the mRNA sequences of the present invention (e.g., modifications to one or both the 3' and 5' ends of an mRNA molecule encoding a functional secreted protein or enzyme). Such modifications include the addition of bases to an mRNA sequence (e.g., the inclusion of a poly A tail or a longer poly A tail), the alteration of the 3' UTR or the 5' UTR, complexing the mRNA with an agent (e.g., a protein or a complementary nucleic acid molecule), and inclusion of elements which change the structure of an mRNA molecule (e.g., which form secondary structures). [0223] The poly A tail is thought to stabilize natural messengers. Therefore, in one embodiment a long poly A tail can be added to an mRNA molecule thus rendering the mRNA more stable. Poly A tails can be added using a variety of art-recognized techniques. For example, long poly A tails can be added to synthetic or in vitro transcribed mRNA using poly A polymerase (Yokoe, et al. Nature Biotechnology. 1996; 14: 1252-1256, the contents of which is hereby incorporated by reference herein in its entirety). A transcription vector can also encode long poly A tails. In addition, poly A tails can be added by transcription directly from PCR products. In one embodiment, the length of the poly A tail is at least about 90, 200, 300, 400 at least 500 nucleotides. In one embodiment, the length of the poly A tail is adjusted to control the stability of a modified mRNA molecule of the invention and, thus, the transcription of protein. For example, since the length of the poly A tail can influence the half-life of an mRNA molecule, the length of the poly A tail can be adjusted to modify the level of resistance of the mRNA to nucleases and thereby control the time course of protein expression in a cell. In one embodiment, the stabilized mRNA molecules are sufficiently resistant to in vivo degradation (e.g., by nucleases), such that they may be delivered to the target cell without a transfer vehicle. [0224] In some embodiment embodiments, an mRNA can be modified by the incorporation 3' and/or 5' untranslated (UTR) sequences which are not naturally found in the wild-type mRNA. In one embodiment, 3' and/or 5' flanking sequence which naturally flanks an mRNA and encodes a second, unrelated protein can be incorporated into the nucleotide sequence of an mRNA molecule encoding a therapeutic or functional protein in order to modify it. For example, 3' or 5' sequences from mRNA molecules which are stable (e.g., globin, actin, GAPDH, tubulin, histone, or citric acid cycle enzymes) can be incorporated into the 3' and/or 5' region of a sense mRNA nucleic acid molecule to increase the stability of the sense mRNA molecule. See, e.g., US2003/0083272, the contents of which is hereby incorporated by reference herein in its entirety. More detailed descriptions of the mRNA modifications can be found in US2017/0210698A1, at pages 57-68, which content is incorporated herein by reference in its entirety. H. Template nucleic acid [0225] The compositions, preparations, nanoparticles, and/or nanomaterials and methods disclosed herein may include a template nucleic acid. A template may be used to alter or insert a nucleic acid sequence at or near a target site for an RNA-guided DNA binding protein such as a Cas nuclease, e.g., a Class 2 Cas nuclease. In some embodiments, the methods comprise introducing a template to the cell. In some embodiments, a single template may be provided. In some embodiments, two or more templates may be provided such that editing may occur at two or more target sites. For example, different templates may be provided to edit a single gene in a cell, or two different genes in a cell. [0226] In some embodiments, a template may be used in homologous recombination. In some embodiments, the homologous recombination may result in the integration of the template sequence or a portion of the template sequence into the target nucleic acid molecule. In some embodiments, a template may be used in homology-directed repair, which involves DNA strand invasion at the site of the cleavage in the nucleic acid. In some embodiments, homology-directed repair may result in including the template sequence in the edited target nucleic acid molecule. In some embodiments, a template may be used in gene editing mediated by non-homologous end joining. In some embodiments, a template sequence has no similarity to the nucleic acid sequence near the cleavage site. In some embodiments, a template or a portion of the template sequence is incorporated. In some embodiments, a template includes flanking inverted terminal repeat (ITR) sequences. [0227] In some embodiments, a template sequence may correspond to, comprise, or consist of an endogenous sequence of a target cell. It may also or alternatively correspond to, comprise, or consist of an exogenous sequence of a target cell. As used herein, the term “endogenous sequence” refers to a sequence that is native to the cell. The term “exogenous sequence” refers to a sequence that is not native to a cell, or a sequence whose native location in the genome of the cell is in a different location. In some embodiments, the endogenous sequence may be a genomic sequence of the cell. [0228] In some embodiments, the endogenous sequence may be a chromosomal or extrachromosomal sequence. In some embodiments, an endogenous sequence may be a plasmid sequence of the cell. [0229] In some embodiments, a template contains ssDNA or dsDNA containing flanking invert- terminal repeat (ITR) sequences. In some embodiments, a template is provided as a vector, plasmid, minicircle, nanocircle, or PCR product. [0230] In some embodiments, a nucleic acid is purified. In some embodiments, a nucleic acid is purified using a precipitation method (e.g., LiCl precipitation, alcohol precipitation, or an equivalent method, e.g., as described herein). In some embodiments, a nucleic acid is purified using a chromatography-based method, such as an HPLC-based method or an equivalent method (e.g., as described herein). In some embodiments, a nucleic acid is purified using both a precipitation method (e.g, LiCl precipitation) and an HPLC-based method. In some embodiments, the nucleic acid is purified by tangential flow filtration (TFF). IV. Methods of manufacturing LNPs [0231] Methods of manufacturing lipid nanoparticles are known in the art. In some embodiments, the described compositions, preparations, nanoparticles, and/or nanomaterials are manufactured using microfluidics. For instance, exemplary methods of using microfluidics to form lipid nanoparticles are described by Leung, A.K.K, et al., J Phys Chem, 116:18440-18450 (2012), Chen, D., et al., J Am Chem Soc, 134:6947-6951 (2012), and Belliveau, N.M., et al., Molecular Therapy- Nucleic Acids, 1: e37 (2012), the disclosures of which are hereby incorporated by reference in their entireties. [0232] Briefly, a cargo, such as a cargo described herein, is prepared in a first buffer solution. The other lipid nanoparticle components (such as ionizable lipid, conjugate-linker lipids, cholesterol, and phospholipid) are prepared in a second buffer solution. In some embodiments, a syringe pump introduces the two solutions into a microfluidic device. The two solutions come into contact within the microfluidic device to form lipid nanoparticles encapsulating the cargo. [0233] Methods of screening the disclosed lipid nanoparticles are described in International Patent Application No. PCT/US2018/058171, which is incorporated by reference in its entirety herein. In some embodiments, the screening methods characterize vehicle delivery preparations to identify preparations with a desired tropism and that deliver functional cargo to the cytoplasm of specific cells. In some embodiments, the screening method uses a reporter that has a functionality that can be detected when delivered to the cell. For example, detecting a functional reporter in a cell indicates that the LNP preparation delivers functional cargo to the cell. Among other things, in some embodiments, a chemical composition identifier is included in each different delivery vehicle formulation to keep track of the chemical composition specific for each different delivery vehicle formulation. In some embodiments, a chemical composition identifier is a nucleic acid barcode. In some embodiments, a sequence of the nucleic acid barcode is paired to which chemical components were used to formulate the LNP preparation in which it is loaded so that when the nucleic acid barcode is sequenced, the chemical composition of the delivery vehicle that delivered the barcode is identified. Representative barcodes include, but are not limited to, barcodes described by Sago, 2018 PNAS, Sago, JACS 2018, the disclosure of which is hereby incorporated by reference in its entirety. Representative reporters include, but are not limited to siRNA, mRNA, nuclease protein, nuclease mRNA, small molecules, epigenetic modifiers, and phenotypic modifiers. DNA (genomic and DNA barcodes) can be isolated using QuickExtract (Lucigen) and sequenced using Illumina MiniSeq as described by Sago et al. PNAS 2018, Sago et al. JACs 2018, Sago, Lokugamage et al. Nano Letters 2018, the disclosures of which are hereby incorporated by reference in their entireties). V. Methods of use [0234] Among other things, the present disclosure describes methods of using compositions, preparations, nanoparticles, and/or nanomaterials described herein. For example, in some embodiments, the present disclosure describes methods of using compositions, preparations, nanoparticles, and/or nanomaterials to deliver cargo to specific cells, tissues, or organs, as described herein. As another example, in some embodiments, the present disclosure describes methods of treatment and/or delaying and/or arresting progression of a disease or disorder using compositions, preparations, nanoparticles, and/or nanomaterials as described herein. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein are for use in medicine. [0235] In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials described herein deliver therapeutic or prophylactic agents to specific cells or organs in a subject in need thereof. In some embodiments, the compositions, preparations, nanoparticles, and/or nanomaterials deliver therapeutic or prophylactic agents to specific cells or organs in a subject in need thereof in the absence of a targeting ligand. In some embodiments, the compositions, preparations, nanoparticles, and/or nanomaterials are useful to treat or prevent diseases in a subject in need thereof. A. Methods of delivering cargo to cells, tissue, or organs [0236] Among other things, in some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein target a particular type or class of cells (e.g., cells of a particular organ or system thereof), tissues, population of cells, or organs. In some embodiments, the present disclosure provides methods of delivering one or more cargos described herein to a subject in need thereof. In some embodiments, such methods comprise in vivo and/or in vitro delivery. In some embodiments, such methods comprise in vivo delivery. In some embodiments, such methods comprise in vitro delivery. In some embodiments, the present disclosure provides for methods of delivering one or more therapeutic and/or prophylactic nucleic acids to a subject in need thereof are described herein. [0237] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to liver cells in the subject. Exemplary liver cells include but are not limited to hepatocytes. [0238] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to lung cells in the subject. [0239] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to spleen cells in the subject. Exemplary spleen cells include but are not limited to splenic monocytes, splenic T cells, splenic memory B cells, or splenic B cells. [0240] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to bone marrow cells in the subject. Exemplary bone marrow cells include but are not limited to bone marrow monocytes, bone marrow B cells, bone marrow memory B cells, or bone marrow T cells. [0241] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to immune cells in the subject. Exemplary immune cells include but are not limited to CD8+, CD4+, or CD8+CD4+ cells. [0242] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to cells of the central nervous system in the subject. [0243] In some embodiments, a composition, preparation, nanoparticle, and/or nanomaterial comprises a therapeutic and/or prophylactic of interest that may be specifically delivered to hematopoietic stem cells in the subject. Unless otherwise specified, it is understood that the terms “hematopoietic stem cells (HSCs)” and “hematopoietic stem and progenitor cells (HSPCs)” are used interchangeably in the present disclosure. [0244] In some embodiments, the lipid nanoparticles can be formulated to be delivered in the absence of a targeting ligand to a mammalian liver hepatocytes, 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 lipid nanoparticles are delivered to target type or class of cells. In some embodiments, specific delivery may result in a greater than 2 fold, 5 fold, 10 fold, 15 fold, or 20 fold compared to delivery using a conventional nanoparticle system (e.g., MC3-containing LNPs). B. Methods of producing a polypeptide [0245] Among other things, in some embodiments, methods of using compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein are used for methods of producing a polypeptide. Among other things, in some embodiments, lipid nanoparticles described herein can be used for producing a polypeptide in a target cell in a subject in need thereof. For example, in some embodiments, lipid nanoparticles described herein can be used for producing a polypeptide in a target cell in a subject in need thereof. In some embodiments, compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein comprise one or more nucleic sequences to be delivered to a cell. [0246] In some embodiments, one or more nucleic acids are expressed in a cell. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, 5’ cap formation, and/or 3’ end formation); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein. C. Methods of gene regulation [0247] Among other things, in some embodiments, methods of using compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein are used for gene regulation. Among other things, in some embodiments, lipid nanoparticles described herein can be used for reducing and/or increasing gene expression in a target cell in a subject in need thereof. For example, in some embodiments, lipid nanoparticles described herein can deliver one or more nucleic acids to a target cell in the subject without a targeting ligand. In some embodiments, a nucleic acid is an inhibitor nucleic acid. In some embodiments, an inhibitory nucleic acid is an siRNA. In some embodiments, a nucleic acid is a nucleic acid described herein. As another example, in some embodiments, lipid nanoparticles described herein can deliver cargo to a target cell in the subject without a targeting ligand. In some embodiments, cargo is any cargo described herein. [0248] Among other things, in some embodiments, methods of using compositions, preparations, nanoparticles, and/or nanomaterials disclosed herein for editing of a gene in a cell in a subject in need thereof. [0249] In some embodiments, a cell that is targeted for gene regulation is an immune cell. The immune cell can be a T cell, such as CD8+ T cell, CD4+ T cell, or T regulatory cell. Other exemplary immune cells for gene editing include but are not limited to macrophages, dendritic cells, B cells or natural killer cells. In some embodiments, the cell that is targeted for gene regulation in a hepatocyte. [0250] Exemplary genes that can be targeted include but are not limited to T cell receptors, B cell receptors, CTLA4, PD1, FOXO1, FOXO3, AKTs, CCR5, CXCR4, LAG3, TIM3, Killer immunoglobulin-like receptors, GITR, BTLA, LFA-4, T4, LFA-1, Bp35, CD27L receptor, TNFRSF8, TNFRSF5, CD47, CD52, ICAM-1, LFA-3, L-selectin, Ki-24, MB1, B7, B70, M- CSFR, TNFR-II, IL-7R, OX-40, CD137, CD137L, CD30L, CD40L, FasL, TRAIL, CD257, LIGHT, TRAIL-R1, TRAILR2, TRAIL-R4, TWEAK-R, TNFR, BCMA, B7DC, BTLA, B7-H1, B7-H 2 , B7-H3, ICOS, VEGFR2, NKG2D, JAG1, GITR, CD4, CCR2, GATA-3, MTORC1, MTORC2, RAPTOR, GATOR, FOXP3, NFAT, IL2R, and IL7. Other exemplary genes that can be targeted include but are not limited to OCT, G6Pase, Mut, PCCA, PCCB, PCSK9, ALAS1, and PAH. Exemplary tumor-associated antigens that can be recognized by T cells and are contemplated for targeting, include but are not limited to MAGE1, MAGE3, MAGE6, BAGE, GAGE, NYESO- 1, MART1/Melan A, MC1R, GP100, tyrosinase, TRP-1, TRP-2, PSA, CEA, Cyp-B, Her2/Neu, hTERT, MUC1, PRAME, WT1, RAS, CDK-4, MUM-1, KRAS, MSLN and β-catenin. D. Subjects to be treated [0251] In some embodiments, subjects who are treated are mammals experiencing cancer, autoimmune disease, infections disease, organ transplant, organ failure, protein deficiency, or a combination thereof. In some embodiments, a subject is a human. In some embodiments, methods described herein may cause hepatocytes to translate certain proteins. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a hepatocyte. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a splenic T cell. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a splenic B cell. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a splenic monocyte. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a bone marrow cell. In some embodiments, methods described herein may be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to a lung cell. [0252] It should be understood that the order of steps or order for performing certain action is immaterial so long as the invention remains operable. Moreover, two or more steps or actions may be conducted simultaneously. [0253] While the invention has been particularly shown and described with reference to specific preferred embodiments, it should be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Exemplary Embodiments [0254] The following numbered embodiments, while non-limiting, are exemplary of certain aspects of the present disclosure: 1. A compound of formula I: or its N-oxide, or a pharmaceutically acceptable salt thereof, wherein: each of L 1 , L 1’ , and L 1” is independently absent, -C(O)-, or -OC(O)-; each of L 2 , L 2’ , and L 2” is independently absent, an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain, or –(CH 2 ) m -Cy A -(CH 2 ) m’ -; each Cy A is independently an optionally substituted ring selected from phenylene and 3- to 12- membered saturated or partially unsaturated carbocyclylene; each of m and m’ is independently 0, 1, or 2; each of L 3 , L 3’ , and L 3” is independently absent, -C(O)-, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-; each of R 1 , R 1’ , and R 1” is independently – or an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain wherein 1-3 methylene units are optionally and independently replaced with –O- or –NR-; each Cy B is independently an optionally substituted ring selected from 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl; each p is independently 0, 1, 2, or 3; each L 3a is independently absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C1-10 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O- or -NR-; each R a is independently hydrogen or an optionally substituted group selected from C6-20 aliphatic, 3- to 12-membered saturated or partially unsaturated carbocyclyl, 7- to 12-membered saturated or partially unsaturated bridged bicyclyl having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, 1-adamantyl, 2-adamantyl, sterolyl, and phenyl; X 1 is absent, –O–, –S–, or –NR–; X 2 is absent or an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –Cy C -; Cy C is an optionally substituted ring selected from 3- to 7- membered saturated or partially unsaturated carbocyclene, phenylene, 3- to 7-membered saturated or partially unsaturated heterocyclene having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or 5- to 6-membered heteroarylene having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; X 3 is hydrogen or an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl, phenyl, 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and each R is independently hydrogen or an optionally substituted C 1-6 aliphatic group. 2. A compound of embodiment 1, wherein the compound is of Formula I-A: or its N-oxide, or a pharmaceutically acceptable salt thereof. 3. A compound of embodiments 1 or 2, wherein the compound is of Formula I-A-a: or its N-oxide, or a pharmaceutically acceptable salt thereof. 4. A compound of embodiments 1 or 2, wherein the compound is of Formula I-A-b: or its N-oxide, or a pharmaceutically acceptable salt thereof. 5. A compound of any one of embodiments 1, 2, and 4, wherein the compound is of Formula I-A-c:
or its N-oxide, or a pharmaceutically acceptable salt thereof. 6. A compound of embodiments 1 or 2, wherein the compound is of Formula I-B: or its N-oxide, or a pharmaceutically acceptable salt thereof. 7. A compound of any one of embodiments 1, 2, and 6, wherein the compound is of Formula I-B-a: or its N-oxide, or a pharmaceutically acceptable salt thereof. 8. A compound of any one of embodiments 1, 2, and 6, wherein the compound is of Formula I-B-b:
or its N-oxide, or a pharmaceutically acceptable salt thereof. 9. A compound of any one of embodiments 1, 2, 6, and 8, wherein the compound is of Formula I-B-c: or its N-oxide, or a pharmaceutically acceptable salt thereof. 10. A compound of any one of the preceding embodiments, wherein the compound is of Formula I-C: or its N-oxide, or a pharmaceutically acceptable salt thereof. 11. The compound of any one of the preceding embodiments, wherein the compound is a compound of any of Formulae I, I-A, I-A-a, I-A-b, I-A-c, I-B, I-B-a, I-B-b, I-B-c, and I-C, or a pharmaceutically acceptable salt thereof. 12. The compound of any one of embodiments 1-11, wherein L 1 is -C(O)-. 13. The compound of any one of embodiments 1-12, wherein L 1’ is -C(O)-. 14. The compound of any one of embodiments 1-13, wherein L 1” is -C(O)-. 15. The compound of any one of embodiments 1-14, wherein L 2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. 16. The compound of any one of embodiments 1-15, wherein L 2 is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1 - 8 hydrocarbon chain. 17. The compound of any one of embodiments 1-16, wherein L 2 is -CH 2 -, -(CH 2 )4-, -(CH 2 ) 5 -, or -(CH 2 ) 6 -. 18. The compound of any one of embodiments 1-17, wherein L 2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. 19. The compound of any one of embodiments 1-18, wherein L 2’ is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1 - 8 hydrocarbon chain. 20. The compound of any one of embodiments 1-19, wherein L 2’ is -CH 2 -, -(CH 2 ) 4 -, -(CH 2 ) 5 -, or -(CH 2 ) 6 -. 21. The compound of any one of embodiments 1-20, wherein L 2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C 1-12 hydrocarbon chain. 22. The compound of any one of embodiments 1-21, wherein L 2” is an optionally substituted bivalent saturated or unsaturated, straight or branched C1-8 hydrocarbon chain. 23. The compound of any one of embodiments 1-22, wherein L 2” is -CH 2 -, -(CH 2 ) 4 -, -(CH 2 ) 5 - , or -(CH 2 ) 6 -. 24. The compound of any one of embodiments 1-23, wherein L 3 is absent. 25. The compound of any one of embodiments 1-24, wherein L 3’ is absent. 26. The compound of any one of embodiments 1-25, wherein L 3” is absent. 27. The compound of any one of embodiments 1-23, wherein L 3 is -C(O)O-. 28. The compound of any one of embodiments 1-24 and 26-27, wherein L 3’ is -C(O)O-. 29. The compound of any one of embodiments 1-25 and 27-28, wherein L 3” is -C(O)O-. 30. The compound of any one of embodiments 1-29, wherein R 1 is -(CH 2 )p-Cy B . 31. The compound of any one of embodiments 1-29, wherein R 1 is an optionally substituted saturated or unsaturated, straight or branched C 1-20 hydrocarbon chain. 32. The compound of any one of embodiments 1-29 and 31, wherein R 1 is an optionally substituted saturated or unsaturated, straight or branched C 6-12 hydrocarbon chain. 33. The compound of any one of embodiments 1-29, wherein R 1 is , 34. The compound of any one of embodiments 1-33, wherein R 1’ is -(CH 2 ) p -Cy B . 35. The compound of any one of embodiments 1-33, wherein R 1’ is an optionally substituted saturated or unsaturated, straight or branched C1-20 hydrocarbon chain. 36. The compound of any one of embodiments 1-33 and 35, wherein R 1’ is an optionally substituted saturated or unsaturated, straight or branched C 6-12 hydrocarbon chain. 37. The compound of any one of embodiments 1-33, wherein R 1’ is , 38. The compound of any one of embodiments 1-37, wherein R 1” is -(CH 2 )p-Cy B . 39. The compound of any one of embodiments 1-37, wherein R 1” is an optionally substituted saturated or unsaturated, straight or branched C 1-20 hydrocarbon chain. 40. The compound of any one of embodiments 1-37 and 39, wherein R 1” is an optionally substituted saturated or unsaturated, straight or branched C6-12 hydrocarbon chain. 41. The compound of any one of embodiments 1-37, wherein R 1” is , 42. The compound of any one of embodiments 1-41, wherein each Cy B is 1-adamantyl. 43. The compound of any one of embodiments 1-42, wherein each p is independently 0 or 1. 44. The compound of any one of embodiments 1-43, wherein X 1 is –O–. 45. The compound of any one of embodiments 1-43, wherein X 1 is –NR–. 46. The compound of any one of embodiments 1-45, wherein X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1-3 methylene units are optionally and independently replaced with -O-, -NR-, or –Cy C -. 47. The compound of any one of embodiments 1-46, wherein X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C 1-12 hydrocarbon chain, wherein 1 methylene unit is replaced with -NR-. 48. The compound of any one of embodiments 1-47, wherein X 2 is an optionally substituted, bivalent saturated or unsaturated, straight or branched, C4-8 hydrocarbon chain, wherein 1 methylene unit is replaced with -NR-. 49. The compound of any one of embodiments 1-48, wherein X 3 is optionally substituted 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. 50. The compound of any one of embodiments 1-49, wherein X 3 is optionally substituted 5- membered saturated or partially unsaturated heterocyclyl having 1-2 nitrogen atoms. 51. The compound of any one of the preceding embodiments, wherein -X 2 -X 3 is 52. The compound of any one of embodiments 1-51, wherein R is hydrogen. 53. The compound of any one of embodiments 1-51, wherein R is optionally substituted C 1-6 aliphatic. 54. The compound of any one of embodiments 1-51 and 53, wherein R is -CH 2 CH3. 55. The compound of embodiment 1, wherein the compound is selected from Table 1, or a pharmaceutically acceptable salt thereof. 56. A lipid nanoparticle (LNP) preparation comprising an ionizable lipid of any one of embodiments 1-55. 57. A lipid nanoparticle (LNP) preparation comprising: an ionizable lipid of any one of embodiments 1-55; a phospholipid; a sterol; and a conjugate-linker lipid (e.g., polyethylene glycol lipid). 58. The LNP preparation of embodiment 56, further comprising a therapeutic and/or prophylactic agent. 59. The LNP preparation of embodiment 58, wherein the therapeutic and/or prophylactic agent is or comprises one or more nucleic acids. 60. The LNP preparation of embodiment 59, wherein the one or more nucleic acids is or comprises RNA. 61. The LNP preparation of embodiment 59, wherein the one or more nucleic acids is or comprises DNA. 62. The LNP preparation of any one of embodiments 58-61, wherein the LNP preparation is formulated to deliver the therapeutic and/or prophylactic agent to target cells. 63. The LNP preparation of embodiment 62, wherein the target cells are or comprise spleen cells (e.g., splenic B cells, splenic T cells, splenic monocytes), liver cells (e.g., hepatocytes), bone marrow cells (e.g., bone marrow monocytes), immune cells, kidney cells, muscle cells, heart cells, or cells in the central nervous system. 64. The LNP preparation of embodiment 63, wherein the target cells are or comprise hematopoietic stem cells (HSCs). 65. A pharmaceutical composition comprising a LNP preparation of embodiment 56 and a pharmaceutically acceptable excipient. 66. A method for administering a therapeutic and/or prophylactic agent to a subject in need thereof, the method comprising administering the LNP preparation of embodiment 56 or the pharmaceutical composition of embodiment 65 to the subject. 67. A method for treating a disease or a disorder in a subject in need thereof, the method comprising administering the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, to the subject, wherein the therapeutic and/or prophylactic agent is effective to treat the disease. 68. A method for delaying and/or arresting progression a disease or a disorder in a subject in need thereof, the method comprising administering the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, to the subject, wherein the therapeutic and/or prophylactic agent is effective to treat the disease. 69. A method of delivering a therapeutic and/or prophylactic agent to a mammalian cell derived from a subject, the method comprising contacting the cell of the subject having been administered the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65. 70. A method of producing a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, wherein the therapeutic and/or prophylactic agent is or comprises an mRNA, and wherein the mRNA encodes the polypeptide of interest, whereby the mRNA is capable of being translated in the cell to produce the polypeptide of interest. 71. A method of inhibiting production of a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, wherein the therapeutic and/or prophylactic agent is or comprises an RNA, whereby the RNA is capable of inhibiting production of the polypeptide of interest. 72. A method of specifically delivering a therapeutic and/or prophylactic agent to a mammalian organ, the method comprising contacting a mammalian organ with the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, whereby the therapeutic and/or prophylactic agent is delivered to the organ. 73. The method of embodiment 72, comprising administering to a subject the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65, to the subject. 74. A method of vaccinating by administering the LNP preparation of embodiment 56, or the pharmaceutical composition of embodiment 65. 75. A method of inducing an adaptive immune response in a subject, comprising administering to the subject an effective amount of a composition comprising at least one RNA; wherein the composition comprises a LNP preparation comprising a compound of embodiment 1, or a pharmaceutically acceptable salt thereof. Exemplification [0255] The present disclosure exemplifies compositions, preparations, formulations, nanoparticles, and/or nanomaterials described herein. The present disclosure also exemplifies methods of preparing, characterizing, and validating compositions, preparations, formulations, nanoparticles, and/or nanomaterials described herein. Example 1: Materials and Methods [0256] The present Example provides exemplary materials and methods of preparing, characterizing, and validating compositions, preparations, nanoparticles, and/or nanomaterials described herein. LNP preparations [0257] Among other things, the present Example provides for exemplary LNP preparations. [0258] Lipid nanoparticle components are dissolved in 100% ethanol at specified lipid component molar ratios. Nucleic acid (NA) cargo is dissolved in 10 mM citrate, 100 mM NaCl, pH 4.0, resulting in a concentration of NA cargo of approximately 0.22 mg/mL. In some embodiments, NA cargos include both a functional NA and a reporter DNA barcode mixed at mass ratios of 1:10 to 10:1 functional NA to barcode. As described herein, a NA can be a siRNA, an anti-sense, an expressing DNA, or mRNA. [0259] LNPs are prepared with a total lipid to NA mass ratio of 11.7. LNPs are formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Spark or Benchtop series Instruments, according to the manufacturers protocol. A ratio of aqueous to organic solvent of approximately 2:1 or 3:1 is maintained during mixing using differential flow rates. After mixing, LNPs are collected, diluted in PBS (approximately 1:1 v/v). Further buffer exchange is conducted using dialysis in PBS at 4 °C for 4 to 24 hours against a 20kDa filter. After this initial dialysis, each individual LNP preparation is characterized via dynamic light scattering (DLS) to measure the size (e.g., diameter) and polydispersity. In addition, pKa of a subpopulation of LNPs is measured via a 2-(p-toluidino)-6-napthalene sulfonic acid (TNS) assay. LNPs falling within specific diameter and polydispersity ranges are pooled, and further dialyzed against phosphate buffer saline (PBS) at 4°C for 1 to 4 hours against a 100 kDa dialysis cassette. After the second dialysis, LNPs are sterile filtered using 0.22 ^M filter and stored at 4 °C for further use. LNP characterization [0260] DLS - LNP hydrodynamic diameter and polydispersity index (PDI) are measured using high throughput dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt). LNPs are diluted 1X PBS to an appropriate concentration and analyzed. Concentration & Encapsulation Efficiency [0261] Concentration of NA is determined by Qubit microRNA kit (for siRNA) or HS RNA kit (for mRNA) per manufacturer’s instructions. Encapsulation efficiency is determined by measuring nucleic acid concentration in unlysed and lysed LNPs. pKa [0262] A stock solution of 10 mM HEPES (Sigma Aldrich), 10 mM MES (Sigma Aldrich), 10 mM sodium acetate (Sigma), and 140 nM sodium chloride (Sigma Aldrich) is prepared, and pH is adjusted using hydrogen chloride and sodium hydroxide to a range of about pH 4-10. Using four replicates for each pH value, 140 μL pH-adjusted buffer is added to a 96-well plate, followed by the addition 5 μL of 2-(p-toluidino)-6- napthalene sulfonic acid (60 μg/ mL). 5 μL of LNP is added to each well. After 5 min of incubation under gentle shaking, fluorescence is measured using an excitation wavelength of 325 nm and emission wavelength of 435 nm (BioTek Synergy H4 Hybrid). LNP Administration [0263] Male and female mice aged approximately 8-12 weeks are used for the studies described by the present Example. Each mouse is temporarily restrained, and pooled LNP was administered intravenously (IV) via tail vein injection in up to five animals per experiment. Age-matched mice are also used to administer vehicle (1X PBS) via tail vein injection in up to three animals per experiment. At 72 hours post-dose, tissues including liver, spleen, bone marrow, kidney, lung, muscle, and blood are collected for analysis. Flow [0264] Liver, kidney, lung, and muscle tissues are mechanically, and then enzymatically digested using a mixture of proteinases, then passed through a 70 uM filter to generate single cell suspensions. Spleen tissues are mechanically digested to generate single cell suspensions. All tissues are treated with (Ammonium-Chloride-Potassium) ACK buffer to lyse red blood cells, and then stained with fluorescently-labeled antibodies for flow cytometry and fluorescence-activated cell sorting (FACS). Commercially available antibodies are used. Using a BD FACSMelody (Becton Dickinson), samples are acquired via flow cytometry to generate gates prior to sorting. In general, gating structure is size ^ singlet cells ^ live cells ^ cells of interest. T cells are defined as CD45+CD3+, monocytes are defined as CD45+CD11b+, and B cells are defined as CD45+CD19+. Endothelial cells are defined as CD31+, monocytes and Kupffer cells as CD45+CD11b+ and hepatocytes as CD31-/CD45-. For siRNA studies, downregulation of the target gene is gated. For mRNA studies, upregulation of the target gene is gated. Tissues from vehicle-dosed mice are used to set the gates for sorting. Up to 1 million cells of each cell subset with the correct phenotype are sorted into PBS. After sorting, cells are pelleted via centrifugation and DNA is extracted using Quick Extract DNA Extraction Solution (Lucigen) according to manufacturer’s protocol. Following DNA extraction, DNA is stored at -20 °C. Barcoding Sequencing [0265] DNA (genomic and DNA barcodes) are isolated using QuickExtract (Lucigen) and sequenced using Illumina MiniSeq as described herein, normalizing frequency DNA barcode counts in FACS isolated samples to frequency in injected input. These data are plotted as ‘Normalized Fold Above Input’ (data not shown). Confirmation [0266] Structural and functional features of the provided LNPs are confirmed based on protocols described herein. LNP Preparation [0267] Lipid nanoparticle components are dissolved in 100% ethanol at specified lipid component molar ratios. Nucleic acid (NA) cargo is dissolved in 10 mM citrate, 100 mM NaCl, pH 4.0, resulting in a concentration of NA cargo of approximately 0.22 mg/mL. In some embodiments, NA cargos include both a functional NA and a reporter DNA barcode mixed at mass ratios of 1:10 to 10:1 functional NA to barcode. LNPs are formulated with a total lipid to NA mass ratio of 11.7. LNPs are formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Spark or Benchtop series Instruments, according to the manufacturer’s protocol. A 2:1 or 3:1 ratio of aqueous to organic solvent is maintained during mixing using differential flow rates. After mixing, LNPs are collected, diluted in PBS (approximately 1:1 v/v), and further buffer exchange is conducted using dialysis in PBS at 4 °C for 8 to 24 hours against a 20 kDa filter. After this initial dialysis, each individual LNP formulation is characterized via DLS to measure the size and polydispersity, and the pKa of a subpopulation of LNPs is measured via TNS assay. After dialysis, LNPs are sterile filtered using 0.22 micron sterile filter and stored at 4 °C for further use. LNP Characterization DLS [0268] LNP hydrodynamic diameter and polydispersity index (PDI) are measured using high throughput dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt). LNPs are diluted 1X PBS to an appropriate concentration and analyzed. Concentration & Encapsulation Efficiency [0269] Concentration of NA is determined by Qubit microRNA kit (for siRNA) or HS RNA kit (for mRNA) per manufacturer’s instructions. Encapsulation efficiency is determined by measuring unlysed and lysed LNPs. pKa [0270] A stock solution of 10 mM HEPES (Sigma Aldrich), 10 mM MES (Sigma Aldrich), 10 mM sodium acetate (Sigma), and 140 nM sodium chloride (Sigma Aldrich) is prepared and pH adjusted using hydrogen chloride and sodium hydroxide to a range of about pH 4-10. Using four replicates for each pH, 140 μL pH-adjusted buffer is added to a 96-well plate, followed by the addition 5 μL of 2-(p-toluidino)-6- napthalene sulfonic acid (60 μg/ mL). 5 μL of LNP is added to each well. After 5 min of incubation under gentle shaking, fluorescence is measured using an excitation wavelength of 325 nm and emission wavelength of 435 nm (BioTek Synergy H4 Hybrid). LNP Administration [0271] Male and female mice aged approximately 8-12 weeks are used for studies described by the present Example. Each mouse is temporarily restrained, and pooled LNP is administered IV via tail vein injection in up to five animals per experiment. Age-matched mice are also used to administer vehicle (1X PBS) via tail vein injection in up to three animals per experiment. Additional routes of administration can also be conducted including intracerebral ventricular (ICV), intracisterna manga (ICM), intrathecal (IT), intramuscular (IM), nebulization, intranasal (IN), subcutaneous (SC), intraarticular, and intradermal (ID). At 72 hours post-dose, tissues including liver, spleen, bone marrow and blood are collected for analysis. Flow [0272] Liver, kidney, lung, and muscle (e.g. skeletal and cardiac) tissues are mechanically, and then enzymatically digested using a mixture of proteinases, then passed through a 70 uM filter to generate single cell suspensions. Spleen tissues are mechanically digested to generate single cell suspensions. Tissues are treated with ACK buffer to lyse red blood cells, and then stained with fluorescently-labeled antibodies for flow cytometry and fluorescence-activated cell sorting (FACS). Commercially available antibodies are used in the present example. Using a BD FACSMelody (Becton Dickinson), samples are acquired via flow cytometry to generate gates prior to sorting. In general, the gating structure was size ^ singlet cells ^ live cells ^ cells of interest. T cells are defined as CD45+CD3+, monocytes are defined as CD45+CD11b+, and B cells are defined as CD45+CD19+. Endothelial cells are defined as CD31+, monocytes and Kupffer cells as CD45+CD11b+ and hepatocytes and myocytes are defined as CD31-/CD45-, in the liver and muscle, respectively. Tissues from vehicle-dosed mice are used to set the gates for sorting. hEPO Expression [0273] For human EPO (hEPO) protein expression, mice are temporarily restrained and bled at 6 hours post-administration (via tail vein). Blood is collected in heparin tubes, processed to plasma, and stored at -80 °C until ready to use. Appropriate dilutions of plasma are used to measure hEPO protein using R&D systems ELISA kit (DuoSet; DY286-05) according to manufacturer’s instructions. Tolerability ALT / AST Quantification [0274] For rat Aspartate Transaminase (AST) and Alanine Transaminase (ALT) quantification, rats are temporarily restrained and bled at 2, 4, 6, 24, 48, and 72 hrs hours post-administration. Blood is collected in heparin tubes, processed to plasma, and stored at -80 °C until ready to use. AST is quantified using AST/GOT reagent (ThermoFisher, TR70121) and ALT is quantified using ALT/GPT reagent (ThermoFisher, TR71121) according to manufacturer’s instructions. Rat MCP-1 ELISA [0275] For Rat Monocype Chemoattractant Protein-1 (MCP-1) protein expression, rats are temporarily restrained and bled at 2, 4, 6, 24, 48, and 72 hrs hours post-administration. Blood is collected in heparin tubes, processed to plasma, and stored at -80 °C until ready to use. Appropriate dilutions of plasma are used to measure MCP-1 protein using R&D systems ELISA kit (DuoSet; DY3144-05) according to manufacturer’s instructions. Screening Experiments [0276] As described herein, a plurality of LNPs (for example, more than 300 LNP preparations) can be simultaneously tested in a single screening experiment. In some embodiments, more than 300 LNPs are screened in a single mouse. In some embodiments, more than 850 LNPs are screened in a single mouse (see FIG. 1 and FIG.2). Screening experiments are used to measure mRNA or siRNA delivery to cells and tissues as described herein. [0277] For mRNA delivery, each LNP preparation is formulated to carry Cre mRNA and a barcode as described herein. Each LNP preparation is administered to LSL-tdTom mouse (Ai14) (see FIG. 1) in accordance with the methods described herein (see also FIG.1). Referring to FIG.1, a library of LNP preparations each comprising one or more components, a barcode sequence, and Cre mRNA is administered into a Cre-LoxP reporter mouse. As described herein, mouse cells are sorted using FACS based on whether the cells are tdTom- or tdTom+. Sorted tdTom+ cells are then sequenced as descried herein. [0278] For siRNA delivery, each LNP preparation is formulated to carry siGFP and barcodes, as described herein. Each LNP preparation is administered to a GFP mouse (see FIG. 2) in accordance with the methods described herein (see also FIG.2). Referring to FIG.2, a library of LNP preparations each comprising one or more components, a barcode sequence, and siGFP is administered into a GFP reporter mouse. As described herein, cells are sorted using FACS based on GFP expression. Sorted cells are then sequenced as descried herein. [0279] LNP preparations are formulated using compounds described herein or comparator compounds. LNP preparations are formulated using MC3 as a control. The following measurements are made: LNP preparation diameter, LNP preparation polydispersity, “normalized delivery efficiency” to any combination of cell- and tissue- types, LNP preparation pKA (which is related to, but not the same as lipid pKA), lipid pKA, and LNP preparation ionizability. Encapsulation efficiency and delivery potency are also measured for each pool of LNP preparations. hEPO expression and Cre expression measurements are performed as described herein. Example 2: Potency per screen [0280] The present Example provides exemplary compositions, preparations, nanoparticles, and/or nanomaterials, and materials and methods for screening potency of such compositions, preparations, nanoparticles, and/or nanomaterials described herein. [0281] Exemplary LNP screens as described in Example 1 can be used to demonstrate that each pool of LNPs is highly potent across many tissues. Example 3: Exemplary LNP preparations with potent delivery to various cell types [0282] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials for potent delivery to various cell types described herein. The present example can be used to demonstrate that in some embodiments, provided lipids show potent delivery to various cell types. [0283] Based on the results derived from screens in Example 1, exemplary LNP preparations are identified for use in splenic delivery, and in particular, for use in B cell delivery. These identified lipids are formulated into LNP preparations and screened using a Cre reporter system described herein. Three Ai14 mice are used per group. Payloads comprise 0.3 mg/kg Cre mRNA. Data are collected at 168 hours post-injection. Results are compared to an MC3-LNP preparation as a control. Example 4: Exemplary LNP preparations are delivered to various cell types [0284] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent delivery to various cell types as described herein. The present example can be used to demonstrate that provided lipids show potent delivery across various cell types. [0285] LNP preparations are selected to confirm efficacy results using a Cre reporter system and Ai14 mouse model described herein. Three Ai14 mice per group are used in each experiment. Data is collected 72 hours post-injection. The screening platforms described herein can identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for a particular cell type. Example 5: Exemplary LNP preparations deliver functional mRNA [0286] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that deliver functional mRNA to various cell types. The present example can be used to demonstrate provided lipids are able to deliver functional mRNA in mice. [0287] Exemplary LNP preparations are selected to determine each preparation’s ability to deliver functional mRNA in mice. LNP preparations each carry 0.15 mg/kg hEPO mRNA and are administered at a mass ratio of 11.7 and 19 in 2-3 C57BL6 mice. hEPO expression in plasma is measured six hours post injection of LNP preparations. Example 6: Tolerability and efficacy experiments [0288] The present Example provides exemplary materials and methods of preparing, characterizing, and validating compositions, preparations, nanoparticles, and/or nanomaterials described herein. The present example can be used to demonstrate that provided lipids are able to deliver functional mRNA in mice. [0289] Exemplary LNP preparations are selected to determine tolerability and efficacy of hEPO mRNA delivery in rats as described herein. Each LNP preparation containing 1.0 mg/kg hEPO mRNA is injected into Sprague-Dawley rats (N=2). Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are collected from rat plasma (U/L) at 24 hours post-injection for each exemplary LNP preparation. Monocyte chemoattractant protein-1 (MCP-1) is collected from rat plasma (ng/mL) at 6 hours post-injection for each LNP preparation. Saline is used as a control. hEPO is collected from rat plasma (ng/mL) after administering a vehicle (control) and exemplary LNP preparations across various time points post-injection (0, 2, 4, 6, 24, 48, 96 hours). Example 7: Synthesis of Ionizable lipids [0290] The present Example provides exemplary materials and methods of preparing, characterizing, and validating ionizable lipids as described herein. As described in the Examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It will be appreciated that, although the general methods depict the synthesis of certain compounds of the present disclosure, the following general methods and other methods known to one of ordinary skill in the art can be applied to all compounds and subclasses and species of each of these compounds, as described herein. [0291] General notes: All reactions were run using anhydrous grade solvents under an atmosphere of nitrogen in flasks or vials with magnetic stirring, unless otherwise noted. Anhydrous solvents were purchased from Sigma-Aldrich and used as received. Flash column chromatography was performed using a Biotage Selekt or Teledyne-Isco CombiFlash Nextgen300+ with prepacked silica gel cartridges. Thin layer chromatography was performed using Merck silica gel 60 plates, and compounds were visualized using iodine. Nuclear magnetic resonance (NMR) spectroscopy was performed either using a Varian INOVA 500 MHz or a Bruker AVANCE 400 MHz spectrometer; chemical shifts are reported in ^ parts per million (ppm) referenced to tetramethylsilane at ^ = 0.00 ppm for CDCl3 samples, and residual solvent peak ( ^ = 2.50 ppm) for DMSO samples. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters Acquity UPLC H-class Plus with QDa detector (ESI + ) using one of the following methods: [0292] Method A: 5 min run, Column: XTERRA RP 18 (4.6 x 50 mm), 5 ^m, mobile phase: initially 50% [0.1% HCOOH in water] and 50% [0.1% HCOOH in (70:30) ACN: THF]; then to 2% [0.1% HCOOH in water] and 98% [0.1% HCOOH in (70:30) ACN: THF] in 2.65 min, held this mobile phase composition up to 3.75 min, and finally back to initial condition, i.e; 50% [0.1% HCOOH in water] and 50% [0.1% HCOOH in (70:30) ACN: THF] in 4.90 min, held this mobile phase composition up to 5.10 min. Flow =1.2 mL/min. [0293] Method B: 12 min run, Column: XTERRA RP 18 (4.6 x 50 mm), 5 µm, (mobile phase: initially 80% [0.1% HCOOH in water] and 20% [0.1% HCOOH in (70:30) ACN: THF]; held this initial condition for 0.75 min; then to 65% [0.1% HCOOH in water] and 35% [0.1% HCOOH in (70:30) ACN: THF] in 3.0 min, then to 2% [0.1% HCOOH in water] and 98% [0.1% HCOOH in (70:30) ACN: THF] in 6.0 min, held this mobile phase composition up to 9.0 min, and finally back to initial condition, i.e.; 80% [0.1% HCOOH in WATER] and 20% [0.1% HCOOH in (70:30) ACN: THF] in 11.00 min, held this mobile phase composition up to 12.10 min. Flow =1.2 ml/min List of abbreviations ACN: acetonitrile d: doublet DCC: N,N’-dicyclohexylcarbodiimide DCM: dichloromethane DIPEA: N,N-diisopropylethylamine DMAP: 4-(dimethylamino)pyridine DMSO: dimethyl sulfoxide EDC: N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride Equiv: equivalents Et: ethyl i-Pr: isopropyl m: multiplet Me: methyl p: pentet q: quartet Rt: retention time s: singlet t: triplet TEA: triethylamine THF: tetrahydrofuran Example 7-1: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(((Z)-non-3- en-1-yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate Step 1: (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid General Procedure A: [0294] To a stirred solution of adipic acid (5.1 g, 35 mmol, 5 equiv) in DCM:THF (1:1) (20 mL), were added EDC (1.7 g, 9.1 mmol, 1.3 equiv), DIPEA (2.7 g, 21 mmol, 3.0 equiv) and DMAP (170 mg, 1.4 mmol, 0.2 equiv) at 0 °C. Reaction mixture was stirred for 5 min and then was added (Z)-non-3-en-1-ol (1.0 g, 7.0 mmol, 1.0 equiv). Reaction mixture was stirred at 25 °C for 12 h. Upon completion, reaction mixture was concentrated under reduce pressure, diluted with water (30 mL) and extracted with DCM (3×50 mL). Organic layer was washed with brine (20 mL) and dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduce pressure. The crude material thus obtained was purified by CombiFlash column chromatography, eluted with 0-10% EtOAc- hexane to afford (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid (1.3 g, 69%) as colorless oil. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.86 (t, J = 6.7 Hz, 3H), 1.16 – 1.38 (m, 6H), 1.46 – 1.56 (m, 4H), 2.00 (q, J = 7.1 Hz, 2H), 2.20 (t, J = 6.6 Hz, 2H), 2.24 – 2.35 (m, 4H), 4.00 (t, J = 6.7 Hz, 2H), 5.27 – 5.39 (m, 1H), 5.40 – 5.51 (m, 1H), 12.00 (s, 1H). Step 2: O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate General Procedure B: [0295] To a stirred solution of (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid (406 mg, 1.5 mmol, 3.0 equiv.) in DMF (2 mL), were added EDC (479 mg, 2.5 mmol, 5.0 equiv), DIPEA (388 mg, 3.0 mmol, 6.0 equiv) and DMAP (12 mg, 0.1 mmol, 0.2 equiv). Reaction mixture was stirred at 0 °C for 5 min and then was added 2,2-bis(hydroxymethyl)propane-1,3-diol (68 mg, 0.5 mmol, 1.0 equiv) and stirred at 25 °C for 12 h. Upon completion, reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 30 mL). Organic layers were further washed with cold water (2 x 20 mL) and dried over anhydrous Na2SO4, filtered and concentrated under reduce pressure. Crude material thus obtained was purified by CombiFlash column chromatography, eluted with 9- 10% EtOAc-hexane to afford O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate (100 mg, 23%) as colorless oil. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.8 Hz, 9H), 1.21 – 1.41 (m, 21H), 1.58 – 1.71 (m, 12H), 1.97 – 2.07 (m, 6H), 2.26 – 2.43 (m, 16H), 3.50 (s, 2H), 4.01 – 4.16 (m, 12H), 5.26 – 5.38 (m, 3H), 5.44 – 5.55 (m, 3H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(((Z)-non-3-en-1- yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate (Example 7- 1) General Procedure C: [0296] To a stirred solution of O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate (100 mg, 0.11 mmol, 1 equiv) in dichloromethane (3 mL), were added pyridine (18 mg, 0.22 mmol, 2.0 equiv), 4- nitrophenyl carbonochloridate (45 mg, 0.22 mmol, 2.0 equiv) and DMAP (3 mg, 0.02 mmol, 0.2 equiv). Reaction mixture was stirred at 25 °C for 2 h. Then were added 3-(diethylamino)propan- 1-ol (44 mg, 0.34 mmol, 3.0 equiv) and DIPEA (43 mg, 0.34 mmol, 3.0 equiv) and stirred at 25 °C for 12 h. Upon completion, reaction mixture was diluted with dichloromethane (30 mL), washed with 1M sodium carbonate (3 x 10 mL), dried over anhydrous Na 2 SO 4 and concentrated under reduced pressure. Crude material thus obtained was purified by flash column chromatography, eluted with 5% MeOH in DCM to afford O,O'-(2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)-2-(((6-(((Z)-non- 3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate (53 mg, 45%) as colorless oil. UPLC-MS (Method A): Rt 2.13 min, m/z calculated [M+H]: 1050.7, found 1052.0. Example 7-2: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(nonyloxy)- 6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate Step 1: 6-(nonyloxy)-6-oxohexanoic acid [0297] Prepared according to General Procedure A, substituting nonan-1-ol for (Z)-non-3-en-1-ol. Isolated 1.2 g, 65%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.86 (t, J = 6.6 Hz, 3H), 1.14 – 1.37 (m, 12H), 1.44 – 1.61 (m, 6H), 2.20 (t, J = 6.8 Hz, 2H), 2.28 (t, J = 6.9 Hz, 2H), 3.99 (t, J = 6.6 Hz, 2H), 12.00 (s, 1H).
Step 2: O,O'-(2-(hydroxymethyl)-2-(((6-(nonyloxy)-6-oxohexanoyl)oxy) methyl)propane-1,3-diyl) dinonyl diadipate [0298] Prepared according to General Procedure B, substituting 6-(nonyloxy)-6-oxohexanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 102 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.2 Hz, 9H), 1.16 – 1.39 (m, 35H), 1.44 – 1.74 (m, 20H), 2.25 – 2.43 (m, 12H), 3.50 (s, 2H), 4.04 (t, J = 6.6 Hz, 6H), 4.10 (s, 6H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(nonyloxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate (Example 7-2) [0299] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((6- (nonyloxy)-6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 39 mg, 56%. UPLC-MS (Method A): Rt 2.36 min, m/z c Example 7-3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(((Z)-oct-5- en-1-yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate Step 1: (Z)-6-(oct-5-en-1-yloxy)-6-oxohexanoic acid [0300] Prepared according to General Procedure A, substituting (Z)-oct-5-en-1-ol for (Z)-non-3- en-1-ol. Isolated 1.4 g, 68%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.90 (t, J = 7.4 Hz, 3H), 1.27 – 1.40 (m, 2H), 1.42 – 1.66 (m, 6H), 2.00 (d, J = 6.7 Hz, 4H), 2.20 (t, J = 7.1 Hz, 2H), 2.28 (t, J = 6.8 Hz, 2H), 3.99 (t, J = 6.2 Hz, 2H), 5.18 – 5.47 (m, 2H), 12.01 (s, 1H). Step 2: O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-oct-5-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate [0301] Prepared according to General Procedure B, substituting (Z)-6-(oct-5-en-1-yloxy)-6- oxohexanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 102 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 9H), 1.33 – 1.46 (m, 6H), 1.56 – 1.72 (m, 21H), 1.96 – 2.11 (m, 12H), 2.24 – 2.42 (m, 12H), 3.50 (s, 2H), 4.02 – 4.12 (m, 12H), 5.24 – 5.42 (m, 6H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((6-(((Z)-oct-5-en-1- yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate (Example 7- 3) [0302] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((6- (((Z)-oct-5-en-1-yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1, 3-diyl) di((Z)-oct-5-en-1-yl) diadipate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 54 mg, 65%. UPLC-MS (Method A): Rt 2.07 min, m/z calculated [M+H]: 1008.7, found 1010.0.
Example 7-4: di((Z)-non-3-en-1-yl) O,O'-(2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carba moyl)oxy)methyl)propane- 1,3-diyl) diadipate [0303] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 39 mg, 56%. UPLC-MS (Method A): Rt 2.18 min, m/z c Example 7-5: dinonyl O,O'-(2-(((6-(nonyloxy)-6-oxohexanoyl)oxy)methyl)-2-((((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) diadipate [0304] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O,O'-(2-(hydroxymethyl)-2-(((6-(nonyloxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate for O,O'-(2-(hydroxymethyl)-2- (((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoyl)oxy)methyl)propa ne-1,3-diyl) di((Z)-non-3-en-1- yl) diadipate. Isolated 53 mg, 59%. UPLC-MS (Method A): Rt 2.31 min, m/z calculated [M+H]: 1038.7, found 1040.1.
Example 7-6: di((Z)-oct-5-en-1-yl) O,O'-(2-(((6-(((Z)-oct-5-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carba moyl)oxy)methyl)propane-1,3- diyl) diadipate [0305] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-oct-5-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 37 mg, 47%. UPLC-MS (Method A): Rt 2.04 min, m/z calculated [M+H]: 991.6, found 993.0. Example 7-7: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(((Z)- non-3-en-1-yl)oxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diy l) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [ . I , , Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((7-(((Z)-non-3-en-1-yl)oxy)-7- oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [0307] Prepared according to General Procedure B, substituting (Z)-7-(non-3-en-1-yloxy)-7- oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 152 mg, 23%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.6 Hz, 9H), 1.17 – 1.46 (m, 24H), 1.52 – 1.72 (m, 14H), 2.02 (d, J = 8.0 Hz, 6H), 2.24 – 2.42 (m, 17H), 3.49 (s, 2H), 4.01 – 4.14 (m, 12H), 5.24 – 5.41 (m, 3H), 5.43 – 5.53 (m, 3H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(((Z)-non-3-en-1- yl)oxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) (Example 7-7) [0308] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((7-(((Z)-non-3-en-1-yl)oxy)-7-oxoheptanoyl)oxy)methyl)prop ane-1,3-diyl) 7,7'-di((Z)-non-3- en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 26 mg, 54%. UPLC-MS (Method A): Rt 2.26 min, m/z calculated [M+H]: 1092.8, found 1094.0. Example 7-8: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(nonyloxy)- 7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) Step 1: 7-(nonyloxy)-7-oxoheptanoic acid [0309] Prepared according to General Procedure A, substituting heptanedioic acid for adipic acid and nonan-1-ol for (Z)-non-3-en-1-ol. Isolated 1.4 g, 69%. 1 H NMR (400 MHz, DMSO-d6) δ 0.85 (t, J = 6.6 Hz, 3H), 1.22 – 1.32 (m, 14H), 1.42 – 1.58 (m, 6H), 2.18 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.4 Hz, 2H), 3.99 (t, J = 6.5 Hz, 2H), 11.97 (s, 1H). Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((7-(nonyloxy)-7-oxoheptanoyl)o xy)methyl)propane-1,3- diyl) 7,7'-dinonyl di(heptanedioate) [0310] Prepared according to General Procedure B, substituting 7-(nonyloxy)-7-oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 142 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 9H), 1.18 – 1.42 (m, 40H), 1.56 – 1.70 (m, 21H), 2.25 – 2.37 (m, 12H), 3.50 (s, 2H), 4.04 (t, J = 6.8 Hz, 6H), 4.10 (s, 6H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(nonyloxy)-7- oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) (Example 7-8) [0311] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((7-(nonyloxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-ox ohexanoyl)oxy)methyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 16 mg, 37%. UPLC-MS (Method A): Rt 2.42 min, m/z calculated [M+H]: 1098.8, found 1100.0. Example 7-9: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(((Z)-oct-5- en-1-yl)oxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) Step 1: (Z)-7-(oct-5-en-1-yloxy)-7-oxoheptanoic acid [0312] Prepared according to General Procedure A, substituting heptanedioic acid for adipic acid and (Z)-oct-5-en-1-ol for (Z)-non-3-en-1-ol. Isolated 1.3 g, 66%. 1 H NMR (400 MHz, DMSO-d6) δ 0.91 (t, J = 7.5 Hz, 3H), 1.21 – 1.40 (m, 4H), 1.42 – 1.64 (m, 6H), 2.01 (q, J = 7.9 Hz, 4H), 2.18 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.3 Hz, 2H), 4.00 (t, J = 6.5 Hz, 2H), 5.08 – 5.56 (m, 2H), 11.99 (s, 1H).
Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((7-(((Z)-oct-5-en-1-yl)oxy)-7- oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) [0313] Prepared according to General Procedure B, substituting (Z)-7-(oct-5-en-1-yloxy)-7- oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 132 mg, 20%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 9H), 1.28 – 1.46 (m, 9H), 1.57 – 1.69 (m, 20H), 1.96 – 2.10 (m, 13H), 2.25 – 2.37 (m, 13H), 3.50 (s, 2H), 4.01 – 4.12 (m, 12H), 5.17 – 5.50 (m, 6H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((7-(((Z)-oct-5-en-1- yl)oxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) (Example 7-9) [0314] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((7-(((Z)-oct-5-en-1-yl)oxy)-7-oxoheptanoyl)oxy)methyl)prop ane-1,3-diyl) 7,7'-di((Z)-oct-5-en- 1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 40 mg, 49%. UPLC-MS (Method A): Rt 2.14 min, m/z calculated [M+H]: 1050.7, found 1052.0.
Example 7-10: 7,7'-di((Z)-non-3-en-1-yl) O'1,O1-(2-(((7-(((Z)-non-3-en-1-yl)oxy)-7- oxoheptanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carb amoyl)oxy)methyl)propane- 1,3-diyl) di(heptanedioate) [0315] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((7-(((Z)-non-3-en-1- yl)oxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 36 mg, 51%. UPLC-MS (Method A): Rt 2.23 min, m/z calculated [M+H]: 1075.7, found 1077.2. Example 7-11: 7,7'-dinonyl O'1,O1-(2-(((7-(nonyloxy)-7-oxoheptanoyl)oxy)methyl)-2-((((2 - (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) di(heptanedioate) [0316] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((7-(nonyloxy)-7- oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 47 mg, 51%. UPLC-MS (Method A): Rt 2.37 min, m/z calculated [M+H]: 1081.8, found 1083.1. Example 7-12: 7,7'-di((Z)-oct-5-en-1-yl) O'1,O1-(2-(((7-(((Z)-oct-5-en-1-yl)oxy)-7- oxoheptanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carb amoyl)oxy)methyl)propane- 1,3-diyl) di(heptanedioate) [0317] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((7-(((Z)-oct-5-en-1-yl)oxy)- 7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-ox ohexanoyl)oxy)methyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 65 mg, 56%. UPLC-MS (Method A): Rt 2.05 min, m/z calculated [M+H]: 1033.7, found 1035.1. Example 7-13: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8-(((Z)- non-3-en-1-yl)oxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl ) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) Step 1: (Z)-8-(non-3-en-1-yloxy)-8-oxooctanoic acid [0318] Prepared according to General Procedure A, substituting octanedioic acid for adipic acid. Isolated 550 mg, 52%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.86 (t, J = 6.7 Hz, 3H), 1.19 – 1.37 (m, 11H), 1.40 – 1.57 (m, 4H), 2.00 (q, J = 7.0 Hz, 2H), 2.18 (t, J = 7.4 Hz, 2H), 2.21 – 2.36 (m, 4H), 4.00 (t, J = 6.7 Hz, 2H), 5.27 – 5.38 (m, 1H), 5.40 – 5.52 (m, 1H). Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((8-(((Z)-non-3-en-1-yl)oxy)-8- oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) [0319] Prepared according to General Procedure B, substituting (Z)-8-(non-3-en-1-yloxy)-8- oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 70 mg, 19%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.7 Hz, 9H), 1.21 – 1.38 (m, 34H), 1.49 – 1.70 (m, 16H), 1.97 – 2.09 (m, 6H), 2.22 – 2.42 (m, 19H), 3.48 (s, 2H), 3.97 – 4.20 (m, 6H), 5.26 – 5.39 (m, 2H), 5.42 – 5.55 (m, 2H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8-(((Z)-non-3-en-1- yl)oxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) (Example 7-13) [0320] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((8-(((Z)-non-3-en-1-yl)oxy)-8-oxooctanoyl)oxy)methyl)propa ne-1,3-diyl) 8,8'-di((Z)-non-3-en- 1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 18 mg, 39%. UPLC-MS (Method A): Rt 2.33 min, m/z calculated [M+H]: 1133.8, found 1135.2. Example 7-14: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8- (nonyloxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) Step 1: 8-(nonyloxy)-8-oxooctanoic acid [0321] Prepared according to General Procedure A, substituting octanedioic acid for adipic acid and nonan-1-ol for (Z)-non-3-en-1-ol. Isolated 1.4 g, 67%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.85 (t, J = 6.4 Hz, 3H), 1.22 – 1.32 (m, 16H), 1.41 – 1.61 (m, 6H), 2.18 (t, J = 7.3 Hz, 2H), 2.26 (t, J = 7.2 Hz, 2H), 3.99 (t, J = 6.6 Hz, 2H), 11.95 (s, 1H). Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((8-(nonyloxy)-8-oxooctanoyl)ox y)methyl)propane-1,3- diyl) 8,8'-dinonyl di(octanedioate) [0322] Prepared according to General Procedure B, substituting 8-(nonyloxy)-8-oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 190 mg, 20%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.4 Hz, 9H), 1.15 – 1.41 (m, 48H), 1.47 – 1.70 (m, 19H), 2.22 – 2.37 (m, 12H), 3.49 (s, 2H), 4.04 (t, J = 6.8 Hz, 6H), 4.10 (s, 6H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8-(nonyloxy)-8- oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) (Example 7-14) [0323] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((8-(((Z)-non-3-en-1-yl)oxy)-8-oxooctanoyl)oxy)methyl)propa ne-1,3-diyl) 8,8'-di((Z)-non-3-en- 1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 25 mg, 43%. UPLC-MS (Method A): Rt 2.47 min, m/z calculated [M+H]: 1139.8, found 1141.2. Example 7-15: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8-(((Z)- oct-5-en-1-yl)oxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl ) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) Step 1: (Z)-8-(oct-5-en-1-yloxy)-8-oxooctanoic acid [0324] Prepared according to General Procedure A, substituting octanedioic acid for adipic acid and (Z)-oct-5-en-1-ol for (Z)-non-3-en-1-ol. Isolated 1.9 g, 66%. 1 H NMR (400 MHz, DMSO-d6) δ 0.91 (t, J = 7.4 Hz, 3H), 1.13 – 1.30 (m, 5H), 1.28 – 1.40 (m, 2H), 1.43 – 1.62 (m, 6H), 1.92 – 2.08 (m, 4H), 2.18 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.3 Hz, 2H), 4.00 (t, J = 6.6 Hz, 2H), 5.18 – 5 Step 2: O'1,O1-(2-(hydroxymethyl)-2-(((8-(((Z)-oct-5-en-1-yl)oxy)-8- oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) [0325] Prepared according to General Procedure B, substituting (Z)-8-(oct-5-en-1-yloxy)-8- oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 140 mg, 20%. 1 H NMR (400 MHz, Chloroform-d) δ 0.94 (t, J = 7.6 Hz, 9H), 1.21 – 1.46 (m, 19H), 1.56 – 1.68 (m, 18H), 1.95 – 2.13 (m, 12H), 2.21 – 2.37 (m, 12H), 3.49 (s, 2H), 4.05 (t, J = 6.7 Hz, 6H), 4.10 (s, 6H), 5.10 – 5.55 (m, 6H). Step 3: O'1,O1-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2 -(((8-(((Z)-oct-5-en-1- yl)oxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) (Example 7-15) [0326] Prepared according to General Procedure C, substituting O'1,O1-(2-(hydroxymethyl)-2- (((8-(((Z)-oct-5-en-1-yl)oxy)-8-oxooctanoyl)oxy)methyl)propa ne-1,3-diyl) 8,8'-di((Z)-oct-5-en- 1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 54 mg, 57%. UPLC-MS (Method A): Rt 2.22 min, m/z calculated [M+H]: 1091.8, found 1093.1.
Example 7-16: 8,8'-di((Z)-non-3-en-1-yl) O'1,O1-(2-(((8-(((Z)-non-3-en-1-yl)oxy)-8- oxooctanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carba moyl)oxy)methyl)propane-1,3- diyl) di(octanedioate) [0327] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((8-(((Z)-non-3-en-1- yl)oxy)-8-oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 83 mg, 63%. UPLC-MS (Method A): Rt 2.31 min, m/z calculated [M+H]: 1116.8, found 1118.1. Example 7-17: 8,8'-di((Z)-non-3-en-1-yl) O'1,O1-(2-(((8-(((Z)-non-3-en-1-yl)oxy)-8- oxooctanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carba moyl)oxy)methyl)propane-1,3- diyl) di(octanedioate) [0328] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((8-(nonyloxy)-8- oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 21 mg, 46%. UPLC-MS (Method A): Rt 2.43 min, m/z calculated [M+H]: 1122.8, found 1124.2. Example 7-18: 8,8'-di((Z)-non-3-en-1-yl) O'1,O1-(2-(((8-(((Z)-non-3-en-1-yl)oxy)-8- oxooctanoyl)oxy)methyl)-2-((((2-(pyrrolidin-1-yl)ethyl)carba moyl)oxy)methyl)propane-1,3- diyl) di(octanedioate) [0329] Prepared according to General Procedure C, substituting 2-(pyrrolidin-1-yl)ethan-1-amine for 3-(diethylamino)propan-1-ol and O'1,O1-(2-(hydroxymethyl)-2-(((8-(((Z)-oct-5-en-1-yl)oxy)- 8-oxooctanoyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) for O,O'- (2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexa noyl)oxy)methyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 91 mg, 54%. UPLC-MS (Method A): Rt 2.18 min, m/z calculated [M+H]: 1074.8, found 1076.0. Example 7-19: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) d Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate General Procedure D: [0330] To a stirred solution of (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid (320 mg, 1.2 mmol, 2.0 equiv) in DMF (2 mL), were added EDC (350 mg, 0.6 mmol, 3.0 equiv), DIPEA (310 mg, 2.4 mmol, 4.0 equiv) and DMAP (15 mg, 0.12 mmol, 0.2 equiv) at 0 °C. Reaction mixture was stirred for 5 min and then was added 2,2-bis(hydroxymethyl)propane-1,3-diol (82 mg, 0.6 mmol, 1.0 equiv). The reaction mixture was stirred at 25 °C for 2 h. Upon completion, reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 x 40 mL). Organic layer were washed with brine and dried over anhydrous Na 2 SO 4 , concentrated under reduced pressure. Crude residue thus obtained was purified by CombiFlash column chromatography, eluted with 0-16 % ethyl acetate-hexane to afford O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1- yl) diadipate (80 mg, 21%) as colorless oil. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.9 Hz, 6H), 1.18 – 1.46 (m, 12H), 1.59 – 1.72 (m, 8H), 2.02 (d, J = 7.4 Hz, 4H), 2.29 – 2.39 (m, 12H), 2.63 – 2.81 (m, 2H), 3.57 (s, 4H), 4.06 (t, J = 6.9 Hz, 4H), 4.13 (s, 4H), 5.23 – 5.41 (m, 2H), 5.42 – 5.58 (m, 2H).
Step 2: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-(hy droxymethyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate General Procedure E: [0331] To a stirred solution of 1-adamantaneacetic acid (67 mg, 0.34 mmol, 0.8 equiv) in DCM:THF (1:1) (4 mL), was added EDC (120 mg, 0.64 mmol, 1.5 equiv), DIPEA (110 mg, 0.86 mmol, 2.0 equiv) and DMAP (11 mg, 0.086 mmol, 0.2 equiv) at 0 °C. The reaction mixture was stirred for 5 min at 0°C and then was added O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)- non-3-en-1-yl) diadipate (280 mg, 0.43 mmol, 1.0 equiv). The reaction mixture was stirred for at 25 °C 12 h. Upon completion, reaction mixture was diluted with water (30 mL) and extracted with DCM (3×50 mL). Organic layer were washed with brine and dried over anhydrous Na 2 SO 4 , concentrated under reduced pressure. Crude material thus obtained was purified by CombiFlash column chromatography, eluted with 20-28% EtOAc-hexane to afford O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate (140 mg, 40%) as colorless oil. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.8 Hz, 6H), 1.21 – 1.41 (m, 18H), 1.58 (s, 6H), 1.60 – 1.74 (m, 9H), 1.94 – 1.98 (m, 3H), 2.02 (q, J = 7.1 Hz, 4H), 2.08 (s, 2H), 2.27 – 2.41 (m, 11H), 2.59 (t, J = 6.7 Hz, 1H), 3.51 (d, J = 6.6 Hz, 2H), 4.01 – 4.10 (m, 6H), 4.11 (s, 4H), 5.26 – 5.38 (m, 2H), 5.40 – 5.59 (m, 2H). Step 3: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate (Example 7-19) [0332] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 69 mg, 37%. UPLC-MS (Method A): Rt 2.06 min, m/z calculated [M+H]: 974.7, found 975.0. Example 7-20: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) dinonyl diadipate [0333] Prepared according to General Procedure D, substituting 6-(nonyloxy)-6-oxohexanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 210 mg, 22%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.19 – 1.39 (m, 24H), 1.54 – 1.71 (m, 12H), 2.29 – 2.34 (m, 4H), 2.35 – 2.40 (m, 4H), 2.67 – 2.77 (m, 2H), 3.57 (d, J = 5.0 Hz, 4H), 4.05 (t, J = 6.8 Hz, 4H), 4.13 (s, 4H).
Step 2: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-(hy droxymethyl)propane-1,3- diyl) dinonyl diadipate [0334] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) dinonyl diadipate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 100 mg, 42%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.17 – 1.40 (m, 20H), 1.55 – 1.73 (m, 37H), 1.96 (s, 3H), 2.08 (s, 2H), 2.26 – 2.40 (m, 2H), 3.51 (d, J = 6.7 Hz, 2H), 4.00 – 4.09 (m, 5H), 4.11 (s, 3H). Step 3: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate (Example 7-20) [0335] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) dinonyl diadipate for O,O'- (2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexa noyl)oxy)methyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 61 mg, 57%. UPLC-MS (Method A): Rt 2.18 min, m/z calculated [M+H]: 978.7, found 979.1.
Example 7-21: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate [0336] Prepared according to General Procedure D, substituting (Z)-6-(oct-5-en-1-yloxy)-6- oxohexanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 280 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.94 (t, J = 7.5 Hz, 6H), 1.33 – 1.45 (m, 6H), 1.48 – 1.74 (m, 13H), 1.93 – 2.12 (m, 8H), 2.27 – 2.41 (m, 8H), 3.57 (s, 4H), 4.06 (t, J = 6.7 Hz, 4H), 4.13 (s, 3H), 5.08 – 5.57 (m, 4H).
Step 2: O, O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-(hy droxymethyl)propane- 1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate [0337] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 140 mg, 40%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.7 Hz, 6H), 1.33 – 1.46 (m, 4H), 1.55 – 1.80 (m, 28H), 1.91 – 2.16 (m, 8H), 2.23 – 2.44 (m, 9H), 3.51 (d, J = 6.6 Hz, 2H), 4.03 – 4.13 (m, 11H), 5.17 – 5.54 (m, 4H). Step 3: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate (Example 7-21) [0338] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) di((Z)-oct-5-en-1-yl) diadipate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 31 mg, 58%. UPLC-MS (Method A): Rt 2.02 min, m/z calculated [M+H]: 946.7, found 947.1.
Example 7-22: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (2-(pyrrolidin-1- yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate [0339] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 26 mg, 37%. UPLC-MS (Method A): Rt 2.04 min, m/z calculated [M+H]: 957.7, found 958.2. Example 7-23: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (2-(pyrrolidin- 1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl) dinonyl diadipate [0340] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) dinonyl diadipate for O,O'- (2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexa noyl)oxy)methyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1-yl)ethan-1-amine for 3- (diethylamino)propan-1-ol. Isolated 42 mg, 58%. UPLC-MS (Method A): Rt 2.17 min, m/z calculated [M+H]: 961.7, found 962.2. Example 7-24: O,O'-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-((( (2-(pyrrolidin- 1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diadipate [0341] Prepared according to General Procedure C, substituting O,O'-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) di((Z)-oct-5-en-1-yl) diadipate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 50 mg, 61%. UPLC-MS (Method A): Rt 1.99 min, m/z calculated [M+H]: 929.6, found 930.0. Example 7-25: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate)
Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [0342] Prepared according to General Procedure D, substituting (Z)-7-(non-3-en-1-yloxy)-7- oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 345 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.8 Hz, 6H), 1.18 – 1.45 (m, 16H), 1.58 – 1.71 (m, 8H), 2.0 (q, J = 7.2 Hz, 4H), 2.25 – 2.41 (m, 12H), 2.74 (t, J = 6.4 Hz, 2H), 3.57 (d, J = 6.3 Hz, 4H), 4.05 (t, J = 6.9 Hz, 4H), 4.13 (s, 4H), 5.24-5.42 (m, 2H), 5.43 – 5.55 (m, 2H). Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [0343] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 280 mg, 51%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.7 Hz, 6H), 1.21 – 1.41 (m, 17H), 1.47 – 1.79 (m, 21H), 1.90 – 2.08 (m, 9H), 2.24 – 2.41 (m, 11H), 3.50 (d, J = 6.8 Hz, 2H), 4.01 – 4.13 (m, 10H), 5.26 – 5.40 (m, 2H), 5.43 – 5.53 (m, 2H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) (Example 7-25) [0344] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 73 mg, 61%. UPLC-MS (Method A): Rt 2.18 min, m/z calculated [M+H]: 1002.7, found 1004.1. ( Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) [0345] Prepared according to General Procedure D, substituting 7-(nonyloxy)-7-oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 610 mg, 22%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.5 Hz, 6H), 1.14 – 1.46 (m, 30H), 1.51 – 1.73 (m, 8H), 2.25 – 2.39 (m, 8H), 2.78 (s, 2H), 2.87 (s, 1H), 2.94 (s, 1H), 3.53 – 3.62 (m, 4H), 4.04 (t, J = 6.8 Hz, 4H), 4.13 (s, 4H).
Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 7,7'-dinonyl di(heptanedioate) [0346] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 320 mg, 42%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.24 – 1.41 (m, 29H), 1.46 – 1.75 (m, 22H), 1.96 (s, 3H), 2.08 (s, 2H), 2.25 – 2.37 (m, 8H), 3.50 (s, 2H), 3.69 – 3.78 (m, 1H), 4.00 – 4.13 (m, 10H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) (Example 7-26) [0347] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-dinonyl di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 21 mg, 39%. UPLC-MS (Method A): Rt 2.30 min, m/z calculated [M+H]: 1006.7, found 1008.2.
Example 7-27: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) d Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) [0348] Prepared according to General Procedure D, substituting (Z)-7-(oct-5-en-1-yloxy)-7- oxoheptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 108 mg, 23%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 6H), 1.31 – 1.44 (m, 8H), 1.55 – 1.71 (m, 12H), 2.03 (h, J = 7.2 Hz, 8H), 2.25 – 2.40 (m, 8H), 2.69 – 2.79 (m, 2H), 3.57 (d, J = 5.6 Hz, 4H), 4.05 (t, J = 6.7 Hz, 4H), 4.13 (s, 4H), 5.24 – 5.43 (m, 4H).
Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) [0349] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 180 mg, 48%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 6H), 1.28 – 1.46 (m, 8H), 1.55 – 1.75 (m, 27H), 1.92 – 2.11 (m, 11H), 2.25 – 2.37 (m, 8H), 3.50 (s, 2H), 4.00 – 4.15 (m, 10H), 5.24 – 5.43 (m, 4H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) (Example 7-27) [0350] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 31 mg, 42%. UPLC-MS (Method A): Rt 2.30 min, m/z calculated [M+H]: 974.7, found 976.1.
Example 7-28: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [0351] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 41 mg, 45%. UPLC-MS (Method A): Rt 2.15 min, m/z calculated [M+H]: 985.7, found 987.0. Example 7-29: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 7,7'-dinonyl di(heptanedioate) [0352] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-dinonyl di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 29 mg, 53%. UPLC-MS (Method A): Rt 2.28 min, m/z calculated [M+H]: 989.7, found 991.2. Example 7-30: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) [0353] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 7,7'-di((Z)-oct-5-en-1-yl) di(heptanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 47 mg, 45%. UPLC-MS (Method A): Rt 2.06 min, m/z calculated [M+H]: 957.7, found 959.0. Example 7-31: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) d Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) [0354] Prepared according to General Procedure D, substituting (Z)-8-(non-3-en-1-yloxy)-8- oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 220 mg, 22%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.8 Hz, 6H), 1.23 – 1.40 (m, 22H), 1.57 – 1.68 (m, 6H), 2.02 (q, J = 7.2 Hz, 4H), 2.24 – 2.41 (m, 12H), 2.73 (t, J = 6.4 Hz, 2H), 3.57 (d, J = 6.5 Hz, 4H), 4.05 (t, J = 6.9 Hz, 4H), 4.13 (s, 4H), 5.25 – 5.39 (m, 2H), 5.43 – 5.58 (m, 2H). Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) [0355] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 280 mg, 51%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.5 Hz, 6H), 1.19 – 1.43 (m, 25H), 1.55 – 1.76 (m, 21H), 1.87 – 2.14 (m, 8H), 2.23-2.45 (m, 10H), 3.50 (d, J = 6.7 Hz, 2H), 4.01 – 4.13 (m, 8H), 5.28 – 5.41 (m, 2H), 5.42 – 5.56 (m, 2H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) (Example 7-31) [0356] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 31 mg, 52%. UPLC-MS (Method A): Rt 2.24 min, m/z calculated [M+H]: 1030.7, found 1032.1. Example 7-32: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) [0357] Prepared according to General Procedure D, substituting 8-(nonyloxy)-8-oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 370 mg, 21%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.18 – 1.42 (m, 28H), 1.50 – 1.70 (m, 16H), 2.24 – 2.38 (m, 8H), 2.72 (s, 2H), 3.57 (s, 4H), 4.04 (t, J = 6.7 Hz, 4H), 4.13 (s, 4H). Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 8,8'-dinonyl di(octanedioate) [0358] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 290 mg, 50%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.17 – 1.41 (m, 37H), 1.55 – 1.65 (m, 17H), 1.69 (d, J = 12.5 Hz, 3H), 1.96 (s, 3H), 2.24 – 2.36 (m, 8H), 3.50 (s, 2H), 4.00 – 4.15 (m, 12H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-dinonyl di(octanedioate) (Example 7-32) [0359] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-dinonyl di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 50 mg, 61%. UPLC-MS (Method A): Rt 2.33 min, m/z calculated [M+H]: 1034.7, found 1036.1.
Example 7-33: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) d Step 1: O'1,O1-(2,2-bis(hydroxymethyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) [0360] Prepared according to General Procedure D, substituting (Z)-8-(oct-5-en-1-yloxy)-8- oxooctanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 324 mg, 20%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 6H), 1.29 – 1.46 (m, 12H), 1.56 – 1.68 (m, 12H), 1.96 – 2.11 (m, 8H), 2.24-2.38 (m, 8H), 2.70 (t, J = 6.5 Hz, 2H), 3.57 (d, J = 6.5 Hz, 4H), 4.06 (t, J = 6.7 Hz, 4H), 4.13 (s, 4H), 5.21 – 5.47 (m, 4H).
Step 2: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( hydroxymethyl)propane- 1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) [0361] Prepared according to General Procedure E, substituting O'1,O1-(2,2- bis(hydroxymethyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 204 mg, 52%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 6H), 1.22 – 1.45 (m, 12H), 1.46 – 1.79 (m, 27H), 1.90 – 2.14 (m, 12H), 2.24 – 2.36 (m, 8H), 2.55 (s, 1H), 3.49 (d, J = 6.7 Hz, 2H), 4.01 – 4.13 (m, 8H), 5.23 – 5.46 (m, 4H). Step 3: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) (Example 7-33) [0362] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 73 mg, 61%. UPLC-MS (Method A): Rt 2.09 min, m/z calculated [M+H]: 1002.7, found 1004.0.
Example 7-34: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) [0363] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-di((Z)-non-3-en-1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 87 mg, 62%. UPLC-MS (Method A): Rt 2.16 min, m/z calculated [M+H]: 1013.7, found 1015.0. Example 7-35: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 8,8'-dinonyl di(octanedioate) [0364] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-dinonyl di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 148 mg, 70%. UPLC-MS (Method B): Rt 5.47 min, m/z calculated [M+H]: 1017.7, found 1019.2. Example 7-36: O'1,O1-(2-((2-((3r,5r,7r)-adamantan-1-yl)acetoxy)methyl)-2-( (((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl ) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) [0365] Prepared according to General Procedure C, substituting O'1,O1-(2-((2-((3r,5r,7r)- adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3- diyl) 8,8'-di((Z)-oct-5-en-1-yl) di(octanedioate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 50 mg, 55%. UPLC-MS (Method A): Rt 2.13 min, m/z calculated [M+H]: 985.7, found 987.0. Example 7-37: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(((Z)-non- 3-en-1-yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate S [ . I , 6 – Step 2: O,O'-(2-(hydroxymethyl)-2-(((5-(((Z)-non-3-en-1-yl)oxy)-5- oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate [0367] Prepared according to General Procedure B, substituting (Z)-5-(non-3-en-1-yloxy)-5- oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 250 mg, 20%. 1H NMR (400 MHz, DMSO-d6) δ 0.85 (t, J = 7.0 Hz, 9H), 1.19 – 1.36 (m, 21H), 1.69 – 1.81 (m, 6H), 2.00 (q, J = 6.7 Hz, 6H), 2.25 – 2.39 (m, 18H), 3.99 (t, J = 6.7 Hz, 6H), 4.07 (s, 6H), 5.26 – 5.37 (m, 3H), 5.39 – 5.51 (m, 3H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(((Z)-non-3-en-1- yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate (Example 7-37) [0368] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((5- (((Z)-non-3-en-1-yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1 ,3-diyl) di((Z)-non-3-en-1-yl) diglutarate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 55 mg, 48%. UPLC-MS (Method A): Rt 2.09 min, m/z calculated [M+H]: 1008.7, found 1009.0. Example 7-38: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5- (nonyloxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diglutarate Step 1: 5-(nonyloxy)-5-oxopentanoic acid [0369] Prepared according to General Procedure A, substituting glutaric acid for adipic acid and nonan-1-ol for (Z)-non-3-en-1-ol. Isolated 2.6 g, 67%. 1 H NMR (400 MHz, DMSO-d6) δ 0.86 (t, J = 7.0 Hz, 3H), 1.18 – 1.35 (m, 12H), 1.54 (q, J = 6.8 Hz, 2H), 1.66 – 1.79 (m, 2H), 2.24 (t, J = 7.3 Hz, 2H), 2.31 (t, J = 7.4 Hz, 2H), 4.00 (t, J = 6.6 Hz, 2H), 12.07 (s, 1H). Step 2: O,O'-(2-(hydroxymethyl)-2-(((5-(nonyloxy)-5-oxopentanoyl)oxy )methyl)propane-1,3-diyl) dinonyl diglutarate [0370] Prepared according to General Procedure B, substituting 5-(nonyloxy)-5-oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 290 mg, 23%. 1H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.6 Hz, 9H), 1.09 – 1.45 (m, 36H), 1.54 – 1.65 (m, 6H), 1.86 – 2.02 (m, 6H), 2.31 – 2.44 (m, 12H), 3.51 (d, J = 6.4 Hz, 2H), 4.01 – 4.12 (m, 12H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(nonyloxy)-5- oxopentanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diglutarate (Example 7-38) [0371] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((5- (nonyloxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) dinonyl diglutarate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 75 mg, 54%. UPLC-MS (Method A): Rt 2.27 min, m/z c Example 7-39: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(((Z)-oct- 5-en-1-yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate Step 1: (Z)-5-(oct-5-en-1-yloxy)-5-oxopentanoic acid [0372] Prepared according to General Procedure A, substituting glutaric acid for adipic acid and (Z)-oct-5-en-1-ol for (Z)-non-3-en-1-ol. Isolated 2.3 g, 58%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 0.91 (t, J = 7.4 Hz, 3H), 1.28 – 1.41 (m, 2H), 1.49 – 1.64 (m, 2H), 1.66 – 1.81 (m, 2H), 1.93 – 2.06 (m, 4H), 2.23 (t, J = 7.2 Hz, 2H), 2.32 (t, J = 7.3 Hz, 2H), 4.01 (t, J = 6.4 Hz, 2H), 5.23 – 5.39 (m, 2H), 11.95 – 12.19 (m, 1H).
Step 2: O,O'-(2-(hydroxymethyl)-2-(((5-(((Z)-oct-5-en-1-yl)oxy)-5- oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate [0373] Prepared according to General Procedure B, substituting (Z)-5-(oct-5-en-1-yloxy)-5- oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 235 mg, 20%. 1 H NMR (400 MHz, Chloroform-d) δ 0.95 (t, J = 7.5 Hz, 9H), 1.31 – 1.47 (m, 6H), 1.56 – 1.70 (m, 6H), 1.86 – 2.16 (m, 18H), 2.31 – 2.44 (m, 12H), 2.57 – 2.66 (m, 1H), 3.51 (d, J = 6.8 Hz, 2H), 4.02 – 4.13 (m, 12H), 5.12 – 5.56 (m, 6H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(((Z)-oct-5-en-1- yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate (Example 7-39) [0374] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((5- (((Z)-oct-5-en-1-yl)oxy)-5-oxopentanoyl)oxy)methyl)propane-1 ,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 43 mg, 36%. UPLC-MS (Method A): Rt 1.95 min, m/z calculated [M+H]: 966.6, found 967.0. Example 7-40: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5- (heptyloxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) diheptyl diglutarate
Step 2: diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2- (hydroxymethyl)propane-1,3-diyl) diglutarate [0376] Prepared according to General Procedure B, substituting 5-(heptyloxy)-5-oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 170 mg, 15%. 1H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.3 Hz, 9H), 1.16 – 1.42 (m, 24H), 1.56 – 1.69 (m, 6H), 1.87 – 2.00 (m, 6H), 2.31 – 2.44 (m, 12H), 3.50 (d, J = 6.0 Hz, 2H), 4.05 (t, J = 6.8 Hz, 6H), 4.10 (s, 6H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(heptyloxy)-5- oxopentanoyl)oxy)methyl)propane-1,3-diyl) diheptyl diglutarate (Example 7-40) [0377] Prepared according to General Procedure C, substituting diheptyl O,O'-(2-(((5- (heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(hydroxymethyl)prop ane-1,3-diyl) diglutarate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-ox ohexanoyl)oxy)methyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 27 mg, 37%. UPLC-MS (Method A): Rt 2.03 min, m/z calculated [M+H]: 930.6, found 930.9.
Example 7-41: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(octyloxy)- 5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate Step 1: 5-(octyloxy)-5-oxopentanoic acid [0378] Prepared according to General Procedure A, substituting glutaric acid for adipic acid and octan-1-ol for (Z)-non-3-en-1-ol. Isolated 1.9 g, 50%.1H NMR (400 MHz, DMSO-d 6 ) δ 0.86 (t, J = 6.9 Hz, 3H), 1.22 – 1.33 (m, 8H), 1.34 – 1.44 (m, 1H), 1.50 – 1.61 (m, 2H), 1.66 – 1.79 (m, 2H), 2.23 (t, J = 7.3 Hz, 2H), 2.31 (t, J = 7.4 Hz, 2H), 3.36 (t, J = 6.4 Hz, 1H), 4.00 (t, J = 6.6 Hz, 2H), 12.08 (s, 1H). Step 2: O,O'-(2-(hydroxymethyl)-2-(((5-(octyloxy)-5-oxopentanoyl)oxy )methyl)propane-1,3-diyl) dioctyl diglutarate [0379] Prepared according to General Procedure B, substituting 5-(octyloxy)-5-oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 44 mg, 20%. 1 H NMR (400 MHz, DMSO-d6) δ 0.86 (t, J = 6.5 Hz, 3H), 1.14 – 1.40 (m, 10H), 1.47 – 1.61 (m, 2H), 1.64 – 1.85 (m, 2H), 2.24 (t, J = 7.3 Hz, 2H), 2.31 (t, J = 7.4 Hz, 2H), 4.00 (t, J = 6.6 Hz, 2H), 12.07 (s, 1H). Step 3: O,O'-(2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)-2-( ((5-(octyloxy)-5- oxopentanoyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate (Example 7-41) [0380] Prepared according to General Procedure C, substituting O,O'-(2-(hydroxymethyl)-2-(((5- (octyloxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 44 mg, 37%. UPLC-MS (Method A): Rt 2.12 min, m/z calculated [M+H]: 972.7, found 973.1. Example 7-42: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate [0381] Prepared according to General Procedure D, substituting (Z)-5-(non-3-en-1-yloxy)-5- oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 90 mg, 25%. 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 7.2 Hz, 6H), 1.13 – 1.45 (m, 12H), 1.88 – 2.10 (m, 8H), 2.31 – 2.47 (m, 12H), 2.58 – 2.74 (m, 2H), 3.58 (d, J = 6.2 Hz, 4H), 4.06 (t, J = 7.1 Hz, 4H), 4.13 (s, 4H), 5.18 – 5.68 (m, 4H). Step 2: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) di((Z)- non-3-en-1-yl) diglutarate [0382] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 85 mg, 42%. 1H NMR (400 MHz, Chloroform-d) δ 0.88 (t, J = 6.8 Hz, 6H), 1.22 – 1.39 (m, 17H), 1.55 – 1.75 (m, 15H), 1.90-2.07 (m, 8H), 2.31 – 2.44 (m, 12H), 3.51 (d, J = 6.8 Hz, 2H), 4.02 – 4.15 (m, 8H), 5.15 – 5.67 (m, 4H). Step 3: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate (Example 7-42) [0383] Prepared according to General Procedure C, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-ox ohexanoyl)oxy)methyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 21 mg, 20%. UPLC-MS (Method A): Rt 2.11 min, m/z calculated [M+H]: 946.6, found 947.0. Example 7-43: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dinonyl diglutarate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate [0384] Prepared according to General Procedure D, substituting 5-(nonyloxy)-5-oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 110 mg, 24%. 1H NMR (400 MHz, Chloroform-d) δ 0.83 – 0.92 (m, 6H), 1.17 – 1.41 (m, 24H), 1.56 – 1.69 (m, 4H), 1.85 – 2.05 (m, 4H), 2.29 – 2.53 (m, 8H), 3.58 (s, 4H), 4.05 (t, J = 7.6 Hz, 4H), 4.13 (s, 4H). Step 2: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) dinonyl diglutarate [0385] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diglutarate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 52 mg, 42%. 1H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.18 – 1.38 (m, 26H), 1.53 (s, 6H), 1.58 – 1.74 (m, 8H), 1.89 – 2.00 (m, 7H), 2.08 (s, 2H), 2.31 – 2.44 (m, 8H), 2.57 (t, J = 6.9 Hz, 1H), 3.51 (d, J = 6.8 Hz, 2H), 4.01 – 4.13 (m, 10H). Step 3: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dinonyl diglutarate (Example 7- 43) [0386] Prepared according to General Procedure C, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) dinonyl diglutarate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 20 mg, 32%. UPLC-MS (Method A): Rt 2.19 min, m/z calculated [M+H]: 950.7, found 951.1.
Example 7-44: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate [0387] Prepared according to General Procedure D, substituting (Z)-5-(oct-5-en-1-yloxy)-5- oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 160 mg, 19%. 1H NMR (400 MHz, Chloroform-d) δ 0.94 (t, J = 7.5 Hz, 6H), 1.33 – 1.46 (m, 4H), 1.57 – 1.69 (m, 6H), 1.89 – 2.12 (m, 10H), 2.32 – 2.48 (m, 8H), 2.58 – 2.81 (m, 2H), 3.58 (s, 4H), 4.06 (t, J = 6.7 Hz, 4H), 4.12 (s, 4H), 5.23 – 5.44 (m, 4H). Step 2: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) di((Z)- oct-5-en-1-yl) diglutarate [0388] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 90 mg, 51%. 1H NMR (400 MHz, Chloroform-d) δ 0.94 (t, J = 7.5 Hz, 6H), 1.33 – 1.46 (m, 5H), 1.48 – 1.75 (m, 18H), 1.87 – 2.13 (m, 16H), 2.30 – 2.44 (m, 7H), 3.51 (d, J = 6.2 Hz, 2H), 4.02 – 4.13 (m, 10H), 4.95 – 5.76 (m, 4H). Step 3: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate (Example 7-44) [0389] Prepared according to General Procedure C, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) di((Z)-oct-5-en-1-yl) diglutarate for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-ox ohexanoyl)oxy)methyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 64 mg, 35%. UPLC-MS (Method A): Rt 2.01 min, m/z calculated [M+H]: 918.6, found 919.0. an-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) diheptyl diglutarate Step 1: O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) diheptyl diglutarate [0390] Prepared according to General Procedure D, substituting 5-(heptyloxy)-5-oxopentanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 160 mg, 20%. 1H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.8 Hz, 6H), 1.19 – 1.39 (m, 17H), 1.50 – 1.67 (m, 4H), 1.89 – 2.01 (m, 4H), 2.32 – 2.46 (m, 8H), 3.58 (s, 4H), 3.62 (t, J = 6.6 Hz, 1H), 4.05 (t, J = 6.8 Hz, 4H), 4.12 (s, 4H). Step 2: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) diheptyl diglutarate [0391] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) diheptyl diglutarate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 65 mg, 43%. 1H NMR (400 MHz, Chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H), 1.21 – 1.38 (m, 18H), 1.55 – 1.65 (m, 12H), 1.69 (d, J = 12.0 Hz, 3H), 1.89 – 1.99 (m, 6H), 2.08 (s, 2H), 2.31 – 2.44 (m, 8H), 2.59 (t, J = 6.9 Hz, 1H), 3.51 (d, J = 6.9 Hz, 2H), 4.01 – 4.09 (m, 6H), 4.10 (s, 4H). Step 3: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) diheptyl diglutarate (Example 7- 45) [0392] Prepared according to General Procedure C, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) diheptyl diglutarate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 45 mg, 62%. UPLC-MS (Method A): Rt 2.00 min, m/z calculated [M+H]: 894.6, found 895.1. Example 7-46: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate [ f ( Step 2: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) dioctyl diglutarate [0394] Prepared according to General Procedure E, substituting O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) dioctyl diglutarate for O,O'-(2,2- bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 90 mg, 38%. 1H NMR (400 MHz, Chloroform-d) δ 0.88 (s, 6H), 1.19 – 1.38 (m, 28H), 1.45 – 1.77 (m, 10H), 1.87 – 2.01 (m, 6H), 2.08 (s, 2H), 2.29 – 2.47 (m, 6H), 3.51 (s, 2H), 4.02 – 4.13 (m, 10H) Step 3: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate (Example 7- 46) [0395] Prepared according to General Procedure C, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) dioctyl diglutarate for O,O'-(2- (hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6-oxohexanoy l)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 17 mg, 31%. UPLC-MS (Method A): Rt 2.14 min, m/z calculated [M+H]: 922.6, found 923.0.
Example 7-47: diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(((4- (pyrrolidin-1-yl)butanoyl)oxy)methyl)propane-1,3-diyl) diglutarate General Procedure F: [0396] To a stirred solution of 4-(pyrrolidin-1-yl)butanoic acid (1.0 equiv) in DCM (3 mL/0.07 mmol) were added EDC (1.3 equiv), DMAP (0.2 equiv), and diheptyl O,O'-(2-(((5-(heptyloxy)-5- oxopentanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) diglutarate (1.0 equiv) and anhydrous DIPEA (3.0 equiv). The reaction mixture was stirred at 25 °C for 16 h. Upon completion, the reaction mixture was diluted with DCM (25 mL) and extracted with saturated NaHCO 3 (3x25 mL). Organic layer was washed with water (2x25 mL) and brine (2x15 mL). Organic part was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude material was purified by CombiFlash column chromatography eluted with 10% MeOH- DCM to afford diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(((4-( pyrrolidin- 1-yl)butanoyl)oxy)methyl)propane-1,3-diyl) diglutarate (41 mg, 58%) as colorless liquid. UPLC- MS (Method A): Rt 1.98 min, m/z calculated [M+H]: 912.6, found 913.0. Example 7-48: dioctyl O,O'-(2-(((5-(octyloxy)-5-oxopentanoyl)oxy)methyl)-2-(((4- (pyrrolidin-1-yl)butanoyl)oxy)methyl)propane-1,3-diyl) diglutarate [0397] Prepared according to General Procedure F, substituting O,O'-(2-(hydroxymethyl)-2-(((5- (octyloxy)-5-oxopentanoyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate for diheptyl O,O'-(2- (((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) diglutarate. Isolated 43 mg, 35%. UPLC-MS (Method B): Rt 5.27 min, m/z calculated [M+H]: 954.6, found 955.2. Example 7-49: O,O'-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(((4-(pyrrolid in-1- yl)butanoyl)oxy)methyl)propane-1,3-diyl) dioctyl diglutarate [0398] Prepared according to General Procedure F, substituting O,O'-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) dioctyl diglutarate for diheptyl O,O'-(2- (((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(hydroxymethyl )propane-1,3-diyl) diglutarate. Isolated 33 mg, 59%. UPLC-MS (Method A): Rt 2.15 min, m/z calculated [M+H]: 904.6, found 905.0. Example 7-50: di((Z)-non-3-en-1-yl) O,O'-(2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)-2-(((4-(pyrrolidin-1-yl)butanoyl)oxy )methyl)propane-1,3-diyl) diadipate [0399] Prepared according to General Procedure F, substituting O,O'-(2-(hydroxymethyl)-2-(((6- (((Z)-non-3-en-1-yl)oxy)-6-oxohexanoyl)oxy)methyl)propane-1, 3-diyl) di((Z)-non-3-en-1-yl) diadipate for diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2- (hydroxymethyl)propane-1,3-diyl) diglutarate. Isolated 33 mg, 31%. UPLC-MS (Method B): Rt 5.29 min, m/z calculated [M+H]: 1032.7, found 1033.1.
Example 7-51: 7,7'-di((Z)-non-3-en-1-yl) O'1,O1-(2-(((7-(((Z)-non-3-en-1-yl)oxy)-7- oxoheptanoyl)oxy)methyl)-2-(((4-(pyrrolidin-1-yl)butanoyl)ox y)methyl)propane-1,3-diyl) di(heptanedioate) [0400] Prepared according to General Procedure F, substituting O'1,O1-(2-(hydroxymethyl)-2- (((7-(((Z)-non-3-en-1-yl)oxy)-7-oxoheptanoyl)oxy)methyl)prop ane-1,3-diyl) 7,7'-di((Z)-non-3- en-1-yl) di(heptanedioate) for diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2- (hydroxymethyl)propane-1,3-diyl) diglutarate. Isolated 50 mg, 84%. UPLC-MS (Method A): Rt 2.25 min, m/z calculated [M+H]: 1075.5, found 1075.8. Example 7-52: 7,7'-dinonyl O'1,O1-(2-(((7-(nonyloxy)-7-oxoheptanoyl)oxy)methyl)-2-(((4- (pyrrolidin-1-yl)butanoyl)oxy)methyl)propane-1,3-diyl) di(heptanedioate) [0401] Prepared according to General Procedure F, substituting O'1,O1-(2-(hydroxymethyl)-2- (((7-(nonyloxy)-7-oxoheptanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) for diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(hydro xymethyl)propane-1,3- diyl) diglutarate. Isolated 43 mg, 80%. UPLC-MS (Method A): Rt 2.39 min, m/z calculated [M+H]: 1080.8, found 1080.8.
Example 7-53: O'1,O1-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(((4-(pyrrol idin-1- yl)butanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) [0402] Prepared according to General Procedure F, substituting O'1,O1-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) 7,7'-di((Z)-non-3-en-1-yl) di(heptanedioate) for diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2- (hydroxymethyl)propane-1,3-diyl) diglutarate. Isolated 27 mg, 46%. UPLC-MS (Method A): Rt 2.25 min, m/z calculated [M+H]: 984.7, found 984.8. Example 7-54: O'1,O1-(2-((2-(adamantan-1-yl)acetoxy)methyl)-2-(((4-(pyrrol idin-1- yl)butanoyl)oxy)methyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) [0403] Prepared according to General Procedure F, substituting O'1,O1-(2-((2-(adamantan-1- yl)acetoxy)methyl)-2-(hydroxymethyl)propane-1,3-diyl) 7,7'-dinonyl di(heptanedioate) for diheptyl O,O'-(2-(((5-(heptyloxy)-5-oxopentanoyl)oxy)methyl)-2-(hydro xymethyl)propane-1,3- diyl) diglutarate. Isolated 17 mg, 57%. UPLC-MS (Method A): Rt 2.25 min, m/z calculated [M+H]: 988.7, found 988.8.
Step 1: 6-hydroxyhexyl 2-butyloctanoate General Procedure G: [0404] To a stirred solution of 2-butyloctanoic acid (1 equiv) in DCM (10.0 mL) were added EDC (1.3 equiv), DMAP (0.1 equiv) and DIPEA (1.5 equiv). Reaction mixture was stirred at 25 °C for 10 min and then was added hexane-1,6-diol (3.0 equiv). Reaction mixture was further stirred at 25 °C for 16 h. Water (50 mL) was added in the reaction mass and was extracted with DCM (2 X 100 mL). Organic layer was dried over anhydrous Na 2 SO 4 , evaporated under reduced pressure to get crude compound. Crude compound thus obtained was purified by combi flash chromatography using 10% ethyl acetate in hexanes to afford 6-hydroxyhexyl 2-butyloctanoate (1.0 g, 67%) as colorless oil. 1 H NMR (400 MHz, Chloroform-d) δ 0.80 – 0.90 (m, 6H), 1.10 – 1.70 (m, 25H), 2.26 – 2.33 (m, 1H), 3.61 – 3.66 (m, 2H), 4.06 (t, J = 6.6 Hz, 2H). Step 2: 6-((2-butyloctanoyl)oxy)hexanoic acid General Procedure H: [0405] To a stirred solution of 6-hydroxyhexyl 2-butyloctanoate (1.0 equiv) in acetone (10 mL) was added Jones reagent (2M solution, 1.2 equiv) dropwise under ice cold condition. The cooling bath was removed, and stirring was continued 2 h. Upon completion was added isopropanol (3 mL) and the mixture was filtered. The filtrate was concentrated under reduced pressure. The crude mass was diluted with water and the pH of the aqueous layer was adjusted to 6 and the mixture was extracted with 5% MeOH in DCM (3 x 50 mL). The combined dichloromethane extracts were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to afford 6-((2-butyloctanoyl)oxy)hexanoic acid (700 mg, 67%) as light green oil. Step 3: ((2,2-bis(hydroxymethyl)propane-1,3-diyl)bis(oxy))bis(6-oxoh exane-6,1-diyl) bis(2- butyloctanoate) [0406] Prepared according to General Procedure D, substituting 6-((2- butyloctanoyl)oxy)hexanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 160 mg, 20%. 1 H NMR: (400 MHz, Chloroform-d) δ 0.78 – 0.87 (m, 12H), 1.13 – 1.70 (m, 40H), 2.26 – 2.37 (m, 6H), 2.66 – 2.69 (m, 2H), 2.87 (s, 2H), 2.94 (s, 2H), 3.56 – 3.57 (m, 4H), 4.04 – 4.07 (m, 4H), 4.13 (s, 4H). Step 4: 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanenitrile [0407] To a vial containing pyridinium p-toluene sulfonate (0.12 g, 0.48 mmol, 0.05 equiv) was added 4,4-diethoxybutanenitrile (1.5 g, 9.5 mmol, 1 equiv) and cis-5-octen-1-ol (3.7 g, 29 mmol, 3 equiv). The vial was tightly capped, and the resulting mixture was heated at 105 ºC for 72 h. After this time, the mixture was allowed to cool to room temperature. The crude material was purified by flash column chromatography (100 g silica, 0 to 100% dichloromethane in hexanes over 20 minutes). Isolated 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanenitrile (1.14 g, 37%) as a colorless oil. 1 H NMR (500 MHz, Chloroform-d) δ 5.43 – 5.27 (m, 4H), 4.56 (t, J = 5.3 Hz, 1H), 3.61 (dt, J = 9.3, 6.6 Hz, 2H), 3.44 (dt, J = 9.3, 6.6 Hz, 2H), 2.42 (t, J = 7.4 Hz, 2H), 2.12 – 1.91 (m, 9H), 1.66 – 1.54 (m, 5H), 1.49 – 1.36 (m, 4H), 0.96 (t, J = 7.6 Hz, 6H). Step 5: 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid [0408] To a vial containing 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanenitrile (1.14 g, 3.54 mmol, 1 equiv) was added potassium hydroxide (0.60 g, 10.6 mmol, 3 equiv) followed by ethanol (3.5 mL) and water (3.5 mL). The vial was tightly capped, and the reaction mixture was heated to 110 ºC for 18 h. After this time, the mixture was allowed to cool to room temperature. The mixture was diluted with ethyl acetate (20 mL), and the pH was adjusted to ~5 by the addition of 1M HCl. The resulting biphasic mixture was separated, and the aqueous phase was extracted two more times with ethyl acetate (2 x 20 mL). The organic extracts were combined, dried over sodium sulfate, filtered and concentrated to afford 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid (1.16 g, 96% yield) as a sticky white solid. 1 H NMR (400 MHz, Chloroform-d) δ 5.41 – 5.24 (m, 4H), 4.45 (t, J = 5.6 Hz, 1H), 3.51 (dt, J = 9.0, 6.7 Hz, 2H), 3.39 (dt, J = 9.0, 6.7 Hz, 2H), 2.17 (t, J = 7.6 Hz, 2H), 2.08 – 1.98 (m, 8H), 1.81 (q, J = 7.3 Hz, 2H), 1.59 – 1.52 (m, 4H), 1.44 – 1.32 (m, 4H), 0.94 ( Step 6: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -(hydroxymethyl)propane- 1,3-diyl)bis(oxy))bis(6-oxohexane-6,1-diyl) bis(2-butyloctanoate) [0409] Prepared according to General Procedure E, substituting ((2,2- bis(hydroxymethyl)propane-1,3-diyl)bis(oxy))bis(6-oxohexane- 6,1-diyl) bis(2-butyloctanoate) for O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 4,4- bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid for 1-adamantaneacetic acid. Isolated 90 mg, 34%. 1 H NMR : (400 MHz, Chloroform-d) δ 0.85 – 0.96 (m, 14H), 1.24 – 1.64 (m, 38H), 1.91 – 2.58 (m, 16H), 3.36 – 3.56 (m, 5H), 4.03 – 4.09 (m, 8H), 4.45 – 4.47 (m, 1H), 5.28 – 5.36 (m, 4H). Step 7: Example 7-55 [0410] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(6-oxohexane-6,1- diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 62 mg, 56%. 1 H NMR (400 MHz, Chloroform-d) δ 0.85 – 1.65 (m, 81H), 1.87 – 1.60 (m, 22H), 3.36 – 3.58 (m, 4H), 4.03 – 4.17 (m, 13H), 4.45 – 4.47 (m, 1H), 5.26 – 5.39 (m, 4H). UPLC-MS (Method A): Rt 2 [ ( Step 2: 7-((2-butyloctanoyl)oxy)heptanoic acid [0412] Prepared according to General Procedure H, substituting 7-hydroxyheptyl 2- butyloctanoate for 6-hydroxyhexyl 2-butyloctanoate. Isolated 800 mg, 66%. 1 H NMR (400 MHz, DMSO-d6) δ 0.82 – 0.85 (m, 6H), 1.19 – 1.60 (m, 24H), 2.16 – 2.31 (m, 3H), 4.00 (t, J = 6.3 Hz, 2 Step 3: ((2,2-bis(hydroxymethyl)propane-1,3-diyl)bis(oxy))bis(7-oxoh eptane-7,1-diyl) bis(2- butyloctanoate) [0413] Prepared according to General Procedure D, substituting 7-((2- butyloctanoyl)oxy)heptanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 170 mg, 19%. 1 H NMR (400 MHz, Chloroform-d) δ 0.87 – 0.91 (m, 12H), 1.23 – 1.64 (m, 47H), 2.30 – 2.39 (m, 6H), 2.76 (t, J = 6.4 Hz, 2H), 2.89 – 2.97 (m, 1H), 3.58 – 3.60 (m, 4H), 4.06 – 4.16 (m, 8 Step 4: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -(hydroxymethyl)propane- 1,3-diyl)bis(oxy))bis(7-oxoheptane-7,1-diyl) bis(2-butyloctanoate) [0414] Prepared according to General Procedure E, substituting ((2,2- bis(hydroxymethyl)propane-1,3-diyl)bis(oxy))bis(7-oxoheptane -7,1-diyl) bis(2-butyloctanoate) for O,O'-(2,2-bis(hydroxymethyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 4,4- bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid for 1-adamantaneacetic acid. Isolated 85 mg, 33%. 1 H NMR :(400 MHz, Chloroform-d) δ 0.84 – 0.96 (m, 17H), 1.13 – 1.71 (m, 46H), 1.91 – 2.59 (m, 18H), 3.36 – 3.56 (m, 6H), 4.03 – 4.09 (m, 9H), 4.45 – 4.47 (m, 1H), 5.26 – 5.34 (m, 4H). Step 5: Example 7-56 [0415] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(7-oxoheptane- 7,1-diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 60 mg, 54%. UPLC-MS (Method A): Rt 2.36 min, m/z calculated [M+H]: 1236.9, found 1237.3. [ ( Step 2: 8-((2-butyloctanoyl)oxy)octanoic acid [0417] Prepared according to General Procedure H, substituting 8-hydroxyoctyl 2-butyloctanoate for 6-hydroxyhexyl 2-butyloctanoate. Isolated 680 mg, 66%. 1 H NMR (400 MHz, DMSO-d6) δ 0.82 – 0.85 (m, 6H), 1.21 – 1.60 (m, 25H), 2.16 – 2.27 (m, 4H), 4.01 (t, J = 6.2 Hz, 2H), 11.95 (s, 1H).
Step 3: 2,2-bis(hydroxymethyl)propane-1,3-diyl bis(8-((2-butyloctanoyl)oxy)octanoate) [0418] Prepared according to General Procedure D, substituting 8-((2-butyloctanoyl)oxy)octanoic acid for (Z)-6-(non-3-en-1-yloxy)-6-oxohexanoic acid. Isolated 200 mg, 18%. 1 H NMR (400 MHz, Chloroform-d) δ 0.84 – 0.88 (m, 12H), 1.20 – 1.62 (m, 52H), 2.27 – 2.35 (m, 6H), 2.70 – 2.73 (m, 2H), 3.55 – 3.57 (m, 4H), 4.03 – 4.13 (m, 8H). Step 4: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( hydroxymethyl)propane-1,3- diyl bis(8-((2-butyloctanoyl)oxy)octanoate) [0419] Prepared according to General Procedure E, substituting 2,2-bis(hydroxymethyl)propane- 1,3-diyl bis(8-((2-butyloctanoyl)oxy)octanoate) for O,O'-(2,2-bis(hydroxymethyl)propane-1,3- diyl) di((Z)-non-3-en-1-yl) diadipate and 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid for 1- adamantaneacetic acid. Isolated 160 mg, 33%. 1 H NMR (400 MHz, Chloroform-d) δ 0.84 – 0.96 (m, 17H), 1.13 – 1.71 (m, 46H), 1.91 – 2.59 (m, 18H), 3.36 – 3.56 (m, 6H), 4.03 – 4.09 (m, 9H), 4.45 – 4.47 (m, 1H), 5.26 – 5.34 (m, 4H). Step 5: Example 7-57 [0420] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l bis(8-((2- butyloctanoyl)oxy)octanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 60 mg, 56%. UPLC-MS (Method A): Rt 2.40 min, m/z calculated [M+H]: 1265.0, found 1265.4. Example 7-58: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -(((((1- ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)propane-1,3- diyl)bis(oxy))bis(6- oxohexane-6,1-diyl) bis(2-butyloctanoate) [0421] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(6-oxohexane-6,1- diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and (1-ethylpiperidin- 3-yl)methanol for 3-(diethylamino)propan-1-ol. Isolated 62 mg, 56%. UPLC-MS (Method A): Rt 2.30 min, m/z calculated [M+H]: 1220.9, found 1221.2.
Example 7-59: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -(((((1- ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)propane-1,3- diyl)bis(oxy))bis(7- oxoheptane-7,1-diyl) bis(2-butyloctanoate) [0422] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(7-oxoheptane- 7,1-diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and (1-ethylpiperidin- 3-yl)methanol for 3-(diethylamino)propan-1-ol. Isolated 79 mg, 60%. UPLC-MS (Method A): Rt 2.38 min, m/z calculated [M+H]: 1248.9, found 1249.3. Example 7-60: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( ((((1- ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)propane-1,3- diyl bis(8-((2- butyloctanoyl)oxy)octanoate) [0423] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l bis(8-((2- butyloctanoyl)oxy)octanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and (1-ethylpiperidin- 3-yl)methanol for 3-(diethylamino)propan-1-ol. Isolated 52 mg, 53%. UPLC-MS (Method A): Rt 2.40 min, m/z calculated [M+H]: 1277.0, found 1277.3. Example 7-61: [0424] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(6-oxohexane-6,1- diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 3- (ethyl(methyl)amino)propan-1-ol for 3-(diethylamino)propan-1-ol. Isolated 111 mg, 70%. UPLC- MS (Method A): Rt 2.24 min, m/z calculated [M+H]: 1194.9, found 1195.2. [0425] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(7-oxoheptane- 7,1-diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 3- (ethyl(methyl)amino)propan-1-ol for 3-(diethylamino)propan-1-ol. Isolated 81 mg, 60%. UPLC- MS (Method A): Rt 2.33 min, m/z calculated [M+H]: 1222.9, found 1223.3. Example 7-63: [0426] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l bis(8-((2- butyloctanoyl)oxy)octanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 3- (ethyl(methyl)amino)propan-1-ol for 3-(diethylamino)propan-1-ol. Isolated 88 mg, 65%. UPLC- MS (Method A): Rt 2.40 min, m/z calculated [M+H]: 1251.0, found 1251.3.
Example 7-64: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -((((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl )bis(oxy))bis(6-oxohexane- 6,1-diyl) bis(2-butyloctanoate) [0427] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(6-oxohexane-6,1- diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 89 mg, 60%. UPLC-MS (Method A): Rt 2.23 min, m/z calculated [M+H]: 1191.9, found 1192.2. Example 7-65: ((2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2 -((((2- (pyrrolidin-1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl )bis(oxy))bis(7-oxoheptane- 7,1-diyl) bis(2-butyloctanoate) [0428] Prepared according to General Procedure C, substituting ((2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l)bis(oxy))bis(7-oxoheptane- 7,1-diyl) bis(2-butyloctanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 103 mg, 70%. UPLC-MS (Method A): Rt 2.33 min, m/z calculated [M+H]: 1219.9, found 1220.3. Example 7-66: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( (((2-(pyrrolidin- 1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl bis(8-((2-butyloctanoyl)oxy)octanoate) [0429] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l bis(8-((2- butyloctanoyl)oxy)octanoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 100 mg, 70%. UPLC-MS (Method A): Rt 2.36 min, m/z calculated [M+H]: 1248.0, found 1248.4.
( S [ a ( Step 2: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( hydroxymethyl)propane-1,3- diyl (9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) [0431] Prepared according to General Procedure E, substituting 2,2-bis(hydroxymethyl)propane- 1,3-diyl (9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) for O,O'-(2,2-bis(hydroxymethyl)propane- 1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid for 1- adamantaneacetic acid. Isolated 500 mg, 30%. 1 H NMR (400 MHz, Chloroform-d) δ 0.84 – 0.99 (m, 12H), 1.22 – 1.45 (m, 33H), 1.56 – 1.71 (m, 7H), 1.92 (q, J = 7.2 Hz, 2H), 1.96 – 2.09 (m, 16H), 2.30 (t, J = 7.6 Hz, 4H), 2.40 (t, J = 7.4 Hz, 2H), 2.50 – 2.64 (m, 1H), 2.76 (t, J = 6.2 Hz, 4H), 3.33 – 3.44 (m, 2H), 3.45 – 3.63 (m, 4H), 4.09 (s, 6H), 4.47 (t, J = 5.3 Hz, 1H), 5.24 – 5.43 (m, 12H). Step 3: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( (((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propane-1,3-diyl (9Z,9'Z,12Z,12'Z)-bis(octadeca- 9,12-dienoate) (Example 7-67) [0432] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate. Isolated 64 mg, 62%. UPLC-MS (Method A): Rt 2.41 min, m/z calculated [M+H]: 1140.9, found 1141.2. Example 7-68: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( ((((1- ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)propane-1,3- diyl (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) [0433] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and (1-ethylpiperidin- 3-yl)methanol for 3-(diethylamino)propan-1-ol. Isolated 97 mg, 65%. UPLC-MS (Method A): Rt 2.45 min, m/z calculated [M+H]: 1152.9, found 1153.1.
Example 7-69: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( (((3- (ethyl(methyl)amino)propoxy)carbonyl)oxy)methyl)propane-1,3- diyl (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) [0434] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 3- (ethyl(methyl)amino)propan-1-ol for 3-(diethylamino)propan-1-ol. Isolated 79 mg, 61%. UPLC- MS (Method A): Rt 2.45 min, m/z calculated [M+H]: 1126.9, found 1127.1. Example 7-70: 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)methyl)-2-( (((2-(pyrrolidin- 1-yl)ethyl)carbamoyl)oxy)methyl)propane-1,3-diyl (9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12- dienoate) [0435] Prepared according to General Procedure C, substituting 2-(((4,4-bis(((Z)-oct-5-en-1- yl)oxy)butanoyl)oxy)methyl)-2-(hydroxymethyl)propane-1,3-diy l (9Z,9'Z,12Z,12'Z)- bis(octadeca-9,12-dienoate) for O,O'-(2-(hydroxymethyl)-2-(((6-(((Z)-non-3-en-1-yl)oxy)-6- oxohexanoyl)oxy)methyl)propane-1,3-diyl) di((Z)-non-3-en-1-yl) diadipate and 2-(pyrrolidin-1- yl)ethan-1-amine for 3-(diethylamino)propan-1-ol. Isolated 106 mg, 70%. UPLC-MS (Method A): Rt 2.43 min, m/z calculated [M+H]: 1123.9, found 1124.1. Example 8: mRNA delivery of LNP preparations across a variety of cell types [0436] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that deliver functional mRNA to various cell types. The screening platforms described herein can identify several highly potent LNP preparations for delivery of functional mRNA across a variety of cell types. The present example can be used to demonstrate provided lipids are able to deliver functional mRNA in mice, and/or improved delivery (for example, 1.5- 2-fold improvement) of mRNA encoding a reporter to certain cell types. [0437] LNP preparations are selected to determine each preparation’s ability to deliver functional mRNA in mice across a variety of cell types. LNP preparations each contain 1.0 mg/kg reporter mRNA as described herein. A flow assay as described herein is used to measure reporter surface expression at 24 hours post injection of LNP preparations. [0438] Exemplary LNP preparations can provide functional delivery of mRNA encoding a reporter to mice across a variety of HSC cell types (Lin-, Sca-1+, LSK cells, LT-HSCs (CD34- Flk2-), and LT-HSCs (CD150+CD48-)), bone marrow (BM) cell types (Live, T cells, B cells, CD11b, Macrophages, NK cells, CD11c), spleen cell types (T cells, B cells, CD11b, Macrophages, NK cells, CD11c), and liver cell types (CD31, Hepatocytes, CD45, CD11b, Kupffer cells, NK cells, T cells, B cells), with each LNP preparation containing 1.0 mg/kg reporter mRNA compared to a saline control (vehicle). [0439] The present example can be used to demonstrate provided lipids are able to deliver functional mRNA in mice, and/or improved delivery (for example, 1.5-2-fold improvement) of mRNA encoding a reporter to certain cell types. Example 9: mRNA delivery of LNP preparations to Spleen, Liver, and Bone Marrow Cells [0440] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent delivery to various cell types as described herein. The screening platforms described herein can identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for a particular cell type. [0441] The present Example can be used to demonstrate that ionizable lipids effectively deliver mRNA across a variety of cell types in mice. The present example can also be used to demonstrate that provided lipids show potent delivery across various cell types, including bone marrow dendritic cells, bone marrow T cells, splenic B cells, splenic dendritic cells, and LSK (Lin-Sca- 1+c-Kit+) cells, HSPCs, and mouse LT-HSCs. [0442] LNP preparations are selected to confirm efficacy results using a reporter system and C57BL6 mouse models described herein. Three C57BL6 mice per group are used in each experiment. Data are collected 24 hours post-injection. [0443] Heat maps can be used to show average median fluorescence of a variety of LNP preparations containing different ionizable lipids normalized to average median fluorescence of an LNP preparation containing exemplary lipids across a variety of cell types (bone marrow dendritic cells, splenic B cells, and splenic dendritic cells). Each LNP preparation contains 1.0 mg/kg reporter mRNA. [0444] Heat maps can be used to show normalized average % reporter expression or average normalized median fluorescence of a variety of LNP preparations containing different ionizable lipids. Average % reporter and average median fluorescence values are normalized to % reporter expression or average median fluorescence, respectively, of an LNP preparation containing exemplary lipids delivered to T cells (bone marrow cells) or LSK (Lin-Sca-1+c-Kit+) cells. Example 10: mRNA delivery of LNP preparations to Balb/C, C57BL6J mouse strains & humanized (NSG-CD34+) immune mice [0445] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent mRNA delivery to various cell types as described herein. The screening platforms described herein can be used to identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for a particular cell type. The present example can be used to demonstrate that provided lipids show potent delivery across various cell types, including bone marrow live cells, bone marrow CD34+ cells, bone marrow (LSK cells), human LT-HSCs, blood cells (huCD45+), and blood cells (muCD45+). [0446] LNP preparations are selected to confirm efficacy results using a reporter system and a humanized (NSG-CD34+) mouse model or two mice strains (Balb/C, C57BL6J) mice as described herein. Each LNP preparation contains 1.0 mg/kg reporter mRNA. [0447] Exemplary LNP preparations described herein were tested for delivery to blood cells, bone marrow (live cells), and bone marrow (LSK cells) in two mouse strains (Balb/C, C57BL6J mice). [0448] Three Balb/C mice and/or C57BL6J and/or NSG-CD34+ mice per group are used in each experiment. Data are collected 24 hours post-injection. Example 11: LNP preparations for intramuscular mRNA delivery to draining lymph nodes [0449] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that deliver mRNA to draining lymph nodes (e.g., for use as mRNA vaccines). The screening platforms described herein can be used to identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for mRNA delivery to draining lymph nodes (e.g., for use as a mRNA vaccines). Accordingly, the present example can be used to demonstrate that provided lipids show potent delivery to draining lymph nodes. [0450] LNP preparations are selected to determine each preparation’s ability to intramuscularly deliver functional mRNA in Balb/C mice. Each LNP preparation contains 10 µg of Trilink Fluc mRNA. Six hours post injection, skin-draining lymph nodes (dLN) are isolated and luminescence is measured using standard luminescence assays as described herein. Example 12: Utilization of LNP preparations for base editing in spleen, liver, and bone marrow [0451] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that confer gene editing (e.g., using base editors) in a variety of cell types. The screening platforms described herein can be used to identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for base editing of a variety of cell types in vivo (e.g., can perform base editing of cells in mice). The present example can be used to demonstrate that provided lipids can be used to perform base editing in a variety of cell types, including bone marrow, spleen, and liver, and/or improve delivery of LNP preparations to particular cell types. The present example can also be used to demonstrate that mRNA contained with LNP preparations containing an exemplary compound of are more stabilized in the LNP during delivery. Methods [0452] Exemplary LNP preparations are selected to determine each preparation’s ability to perform base editing in a Balb/C mouse model as described herein. [0453] Lipid nanoparticle components are dissolved in 100% ethanol at specified lipid component molar ratios. mRNA encoding an adenine base editor and a chemically-modified sgRNA is dissolved at a mass ratio of 1:1 in 10 mM citrate, 100 mM NaCl, pH 4.0, resulting in a concentration of NA cargo of approximately 0.22 mg/mL. LNP preparations are formulated with molar ratios of 50% Ionizable Lipid : 38.5% Cholesterol : 1.5% PEG2000-DMG: 10% DSPC total lipid to NA mass ratio of 11.7 to 33. LNP preparations are formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Spark or Benchtop series Instruments, according to the manufacturer’s protocol. A 3:1 ratio of aqueous to organic solvent is maintained during mixing using differential flow rates. After mixing, LNP preparations are collected, diluted in PBS (approximately 2:1 v/v), and further buffer exchange is conducted using dialysis in PBS at 4 °C for 8 to 24 hours against a 20 kDa filter. After this initial dialysis, each individual LNP preparation is characterized via dynamic light scattering (DLS) to measure size and polydispersity. pKa of a subpopulation of LNP preparations is measured via TNS assay. After dialysis, LNP preparations are sterile filtered using 0.22 micron sterile filter and stored at 4 °C for further use. In some embodiments, LNP preparations may be concentrated using 100kDa Amicon filters per manufacturer’s protocol. LNP Characterization [0454] DLS - LNP preparation hydrodynamic diameter and polydispersity index (PDI) is measured using high throughput dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt). LNP preparations are diluted 1X PBS to an appropriate concentration and analyzed. Concentration & Encapsulation Efficiency [0455] Concentration of NA is determined by Qubit microRNA kit (for siRNA) or HS RNA kit (for mRNA) per manufacturer’s instructions. Encapsulation efficiency is determined by measuring unlysed and lysed LNPs. LNP Administration [0456] Male Balb/C mice aged approximately 8-12 weeks are used in the experiments described herein. Each mouse is temporarily restrained, and LNP preparations are administered IV via tail vein injection. Age-matched mice are also used to administer vehicle (1X PBS) via tail vein injection as a control. Four to six days post-dose, tissues including liver, spleen, and bone marrow are collected. Genomic DNA is isolated and fragmented and adapter-ligated using the Nextera DNA Flex Library Prep Kit (Illumina) using the 96-well plate Nextera indexing primers (Illumina), according to the manufacturer’s instructions. Library size and concentration is confirmed by Fragment Analyzer (Agilent) and sent to Novogene for whole genome sequencing using an Illumina HiSeq. Base editing [0457] For base editing, mRNA encoding an adenine base editor (ABE) and a guide RNA (sgRNA) targeting ALAS1 at 1:1 (mass ratio) are coencapsulated in LNP preparations as described herein. LNP preparations are administrated into Balb/c mice through tail vein injections at 3.0 mg/kg total RNA. At 4 days after dosing, mice are euthanized and liver, spleen and bone barrow are harvested. Base editing is determined by performing targeted deep sequencing analysis at the ALAS1 target site using extracted genomic DNA. Example 13: Exemplary LNP preparations are delivered to mouse HSPCs [0458] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent delivery to various cell types as described herein. The present example can be used to demonstrate that provided lipids exhibit a dose-dependent increase in particular cell types such as transfected HSPCs. [0459] A LNP preparation is selected to confirm efficacy results using a Cre reporter system and Ai14 mouse model described herein at three different doses (0.1 mg/kg, 0.3 mg/kg, 1.0 mg/kg. Example 14: Exemplary LNP preparations are delivered to mouse HSPCs [0460] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent delivery to various cell types as described herein. [0461] A LNP preparation is selected to confirm efficacy results using a Cre reporter system and Ai14 mouse model described herein by monitoring the durability of % tdTomato+ in HSPCs at multiple timepoints (e.g., at 2, 8, and 16 weeks post LNP delivery at a dose of 0.3 mg/kg Cre mRNA, or at 2, 8, and 16 weeks post LNP delivery at a dose of 1.0 mg/kg Cre mRNA). [0462] When observing durability of the transfected HSPCs, the transfected HSPCs are analyzed to determine their capacity for reconstituting various immune and hematopoietic lineages by measuring the % tdTomato+ abundance in peripheral blood at the 16-week timepoint in monocytes, B cells, T cells, and red blood cells in the mice cohorts administered 0.3 mg/kg and 1.0 mg/kg Cre mRNA. [0463] The present example can be used to demonstrate that provided lipids result in durable LNP delivery to HSPCs. Moreover, the present Example can be used to demonstrate that transfected HSPCs can reconstitute a hematopoietic compartment. Example 15: Exemplary LNP preparations are delivered to human CD34+ HSPCs in vitro [0464] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials with potent delivery to various cell types as described herein. Accordingly, the present example can be used to demonstrate that provided lipids result in increased reporter expression when delivered to particular cell types in the presence of ApoE. [0465] An LNP preparation is selected to confirm efficacy results in human CD34+ HSPCs in vitro. [0466] Briefly, human CD34+ cells are thawed and then plated in a 48-well tissue culture plate at 40,000 cells/well in a cell culture plate. Cells are incubated for 48 hours to reach homeostasis in StemSpan cell culture media (Stem Cell Technologies) supplemented with 10% Penicillin- Streptomycin and StemSpan CD34+ Expansion Supplement (Stem Cell Technologies). Prior to dosing with 1000 ng reporter mRNA encapsulated in the LNP, cells are centrifuged at 300xG and resuspended in supplemented media, as previously described. To test if ApoE was necessary for LNP uptake, 2 ^L of recombinant human apoprotein E3 (ApoE3, R&D Systems) is added to each well of interest. After 24 hours, cell Fc receptors are blocked with human TruStain FcX (Biolegend), washed with PBS, and fluorescently stained for reporter expression along with phenotyping markers and viability dye. Example 16: LNP preparation and characterization [0467] Among other things, the present Example provides for exemplary LNP preparations. [0468] Lipid nanoparticle components were dissolved in 100% ethanol at specified lipid component molar ratios. Nucleic acid (NA) cargo was dissolved in 10 mM citrate, 100 mM NaCl, pH 4.0, resulting in a concentration of NA cargo of approximately 0.22 mg/mL. In some embodiments, NA cargos include both a functional NA and a reporter DNA barcode mixed at mass ratios of 1:10 to 10:1 functional NA to barcode. As described herein, a NA can be a siRNA, an anti-sense, an expressing DNA, or mRNA. [0469] LNPs were prepared with 47.5% Ionizable Lipid (e.g., a compound provided herein): 40% cholesterol: 2.5% PEG-2000 DMG: and 10% DPSC and an N/P ratio range of 4-8. LNPs were formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Spark or Benchtop series Instruments, according to the manufacturers protocol. A ratio of aqueous to organic solvent of approximately 2:1 or 3:1 was maintained during mixing using differential flow rates. After mixing, LNPs were collected, diluted in phosphate buffer saline (PBS) or Tris-buffered saline (TBS) (approximately 1:1 v/v). Further buffer exchange was conducted using dialysis in PBS or TBS at 4 °C for 4 to 24 hours against a 20 kDa filter. After this initial dialysis, each individual LNP preparation was characterized via dynamic light scattering (DLS) to measure the size (e.g., diameter) and polydispersity. LNPs falling within specific diameter and polydispersity ranges were pooled, and further dialyzed against PBS or TBS at 4 °C for 1 to 4 hours against a 100 kDa dialysis cassette. After the second dialysis, LNPs were sterile filtered using 0.22 µm filter and stored at 4 °C for further use. LNP characterization [0470] LNP hydrodynamic diameter and polydispersity index (PDI) were measured using high throughput dynamic light scattering (DLS) (Stunner, Unchained Labs). LNPs were diluted 1X TBS or PBS to an appropriate concentration and analyzed. [0471] RNA concentration and encapsulation efficiency were determined by Ribogreen assay (BioTek Synergy LX plate reader). [0472] Characterization data for LNPs prepared using provided lipids are summarized in Table 2. Table 2. Example 17: LNP administration and biodistribution [0473] Male and female mice aged approximately 8-12 weeks were used for studies described by the present Example. Each mouse was temporarily restrained and pooled LNP was administered IV via tail vein injection in up to five animals per experiment. Age-matched mice was also used to administer vehicle (1X TBS) via tail vein injection in up to three animals per experiment. Additional routes of administration can also be conducted including intracerebral ventricular (ICV), intracisterna manga (ICM), intrathecal (IT), intramuscular (IM), nebulization, intranasal (IN), subcutaneous (SC), intraarticular, and intradermal (ID). At 4 hours post-dose, tissues including liver, spleen, bone marrow, testes, gastrocnemius, quadricep, lung, heart, kidney, and pancreas may be collected for analysis. Flow [0474] Liver, kidney, lung, and muscle (e.g. skeletal and cardiac) tissues may be mechanically, and then enzymatically digested using a mixture of proteinases, then passed through a 70 µm filter to generate single cell suspensions. Spleen tissues were mechanically digested to generate single cell suspensions. Tissues were treated with ACK buffer to lyse red blood cells, and then stained with fluorescently labeled antibodies for flow cytometry and fluorescence-activated cell sorting (FACS). Commercially available antibodies were used in the present example. Using a BD FACSMelody (Becton Dickinson), samples were acquired via flow cytometry to generate gates prior to sorting. In general, the gating structure was size ^singlet cells ^live cells ^cells of interest. T cells were defined as CD45+CD3+, monocytes are defined as CD45+CD11b+, and B cells are defined as CD45+CD19+. Endothelial cells were defined as CD31+, monocytes and Kupffer cells as CD45+CD11b+ and hepatocytes and myocytes were defined as CD31-/CD45- in the liver and muscle, respectively. Tissues from vehicle-dosed mice were used to set the gates for sorting. [0475] RNA from tissues were extracted using the Reliaprep RNA extraction kit, and libraries were prepared using the RNAseq kit, and sequenced on either a MiSeq or NextSeq500. [0476] To screen for preferential LNP delivery to a target tissues, the relative abundance of each barcode in one or more target tissues was measured and compared to the relative abundance of each barcode to its corresponding relative input abundance to determine fold above input (FAI) for each barcode. If the FAI of a barcode in a target tissue is above a threshold or above the FAI of a different LNP in the same tissue, the LNP can be identified as preferentially delivering to the target tissue. [0477] The FAI is the normalized relative abundance of a barcode in a selected sample as compared to its frequency in the input. The FAI of a barcode indicates how a LNP’s abundance changes relative to the rest of the LNP pool. The FAI value of the barcode was calculated by normalizing the relative abundance in the barcode sequence counts in the isolated samples to its relative abundance in the administration input. For example, the FAI value of 1 represents an LNP appearing at the same frequency in the isolated sample as it does in the administration pool, representing that it displays neutral tropism to the cell-type measured relative to other LNP populations in that same administration pool. The FAI then indicates the performance of an LNP relative to the input LNP composition. [0478] Biodistribution data for LNPs prepared using provided lipids are summarized in Table 3, Table 4, Table 5, and Table 6. Each table describes data obtained from one screening experiment. Table 3. *MC3 is (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino) butanoate, often referred to as DL in-MC3-DMA. Table 4. *MC3 is (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino) butanoate, often referred to as DL in-MC3-DMA. Table 5. *MC3 is (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino) butanoate, often referred to as DL in-MC3-DMA. ND = not determined. Table 6. *MC3 is (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino) butanoate, often referred to as DL in-MC3-DMA. Example 18: Utilization of LNP preparations for base editing in spleen, liver, and bone marrow [0479] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that confer gene editing (e.g., using base editors) in a variety of cell types. The screening platforms described herein can be used to identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for base editing of a variety of cell types in vivo (e.g., can perform base editing of cells in mice). [0480] The present example can be used to demonstrate that provided lipids can be used to perform base editing in a variety of cell types, including bone marrow, spleen, and liver, and/or improve delivery of LNP preparations to particular cell types. The present example can also be used to demonstrate that mRNA contained with LNP preparations containing an exemplary compound of are more stabilized in the LNP during delivery. Methods [0481] Exemplary LNP preparations are selected to determine each preparation’s ability to perform base editing in a Balb/C mouse model as described herein. [0482] Lipid nanoparticle components are dissolved in 100% ethanol at specified lipid component molar ratios. mRNA encoding an adenine base editor and a chemically-modified sgRNA is dissolved at a mass ratio of 1:1 in 50 mM citrate, pH 4.0, resulting in a concentration of NA cargo of approximately 0.1 mg/mL. [0483] LNP preparations are formulated with molar ratios of 47.5% Ionizable Lipid : 40% Cholesterol : 2.5% PEG2000-DMG: 10% DSPC total lipid to NA mass ratio of 6 to 12. [0484] LNP preparations are formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Ignite series Instruments, according to the manufacturer’s protocol. [0485] A 3:1 ratio of aqueous to organic solvent is maintained during mixing using differential flow rates. After mixing, LNP preparations are collected, diluted in TBS (approximately 7:1 v/v), and further buffer exchange is conducted using dialysis in TBS at 4 °C for 8 to 24 hours against a 20 kDa filter. After this initial dialysis, each individual LNP preparation is characterized via dynamic light scattering (DLS) to measure size and polydispersity. pKa of a subpopulation of LNP preparations is measured via TNS assay. [0486] After dialysis, LNP preparations are sterile filtered using 0.22 micron sterile filter and stored at 4 °C for further use. In some embodiments, LNP preparations may be concentrated using 100kDa Amicon filters per manufacturer’s protocol. LNP Characterization [0487] DLS - LNP preparation hydrodynamic diameter and polydispersity index (PDI) is measured using high throughput dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt). LNP preparations are diluted 1X PBS to an appropriate concentration and analyzed. Concentration & Encapsulation Efficiency [0488] Concentration of NA is determined by Qubit microRNA kit (for siRNA) or HS RNA kit (for mRNA) per manufacturer’s instructions. Encapsulation efficiency is determined by measuring unlysed and lysed LNPs. LNP Administration [0489] Male Balb/C mice aged approximately 8-12 weeks are used in the experiments described herein. Each mouse is temporarily restrained, and LNP preparations are administrated into Balb/c mice through tail vein injections at 0.15 mg/kg total RNA. [0490] Age-matched mice are also used to administer vehicle (1X TBS) via tail vein injection as a control. Four to six days post-dose, tissues including liver, spleen, and bone marrow are collected. Base editing is determined by performing targeted deep sequencing analysis at the ALAS1 target site using extracted genomic DNA. Genomic DNA is isolated and fragmented and adapter-ligated using the Nextera DNA Flex Library Prep Kit (Illumina) using the 96-well plate Nextera indexing primers (Illumina), according to the manufacturer’s instructions. Library size and concentration is confirmed by Fragment Analyzer (Agilent) and analyzed for whole genome sequencing using an Illumina HiSeq. Example 19: Utilization of LNP preparations for base editing in spleen, liver, and bone marrow [0491] The present Example provides exemplary LNP compositions, preparations, nanoparticles, and/or nanomaterials that confer gene editing (e.g., using base editors) in a variety of cell types. The screening platforms described herein can be used to identify several highly potent LNP preparations to determine what type of LNP preparation would be most potent for base editing of a variety of cell types in vivo (e.g., can perform base editing of cells in mice). [0492] The present example can be used to demonstrate that provided lipids can be used to perform base editing in a variety of cell types, including bone marrow, spleen, and liver, and/or improve delivery of LNP preparations to particular cell types. The present example can also be used to demonstrate that mRNA contained with LNP preparations containing an exemplary compound of are more stabilized in the LNP during delivery. Methods [0493] Exemplary LNP preparations are selected to determine each preparation’s ability to perform base editing in a NBSGW mouse model as described herein. [0494] Lipid nanoparticle components are dissolved in 100% ethanol at specified lipid component molar ratios. mRNA encoding an adenine base editor and a chemically-modified sgRNA is dissolved at a mass ratio of 1:1 in 10 mM citrate, pH 3.0, resulting in a concentration of NA cargo of approximately 0.2 mg/mL. [0495] LNP preparations are formulated with molar ratios of 50% Ionizable Lipid: 38.5% Cholesterol : 1.5% PEG2000-DMG: 10% DSPC total lipid to NA mass ratio of 19 to 1. [0496] LNP preparations are formed by microfluidic mixing of the lipid and NA solutions using a Precision Nanosystems NanoAssemblr Spark series Instruments, according to the manufacturer’s protocol. [0497] A 3:1 ratio of aqueous to organic solvent is maintained during mixing using differential flow rates. After mixing, LNP preparations are collected, diluted in PBS (approximately 2:1 v/v), and further buffer exchange is conducted using dialysis in PBS at 4 °C for 8 to 24 hours against a 20 kDa filter. After this initial dialysis, each individual LNP preparation is characterized via dynamic light scattering (DLS) to measure size and polydispersity. pKa of a subpopulation of LNP preparations is measured via TNS assay. [0498] After dialysis, LNP preparations are sterile filtered using 0.22 micron sterile filter and stored at 4 °C for further use. In some embodiments, LNP preparations may be concentrated using 100 kDa Amicon filters per manufacturer’s protocol. LNP Characterization [0499] DLS - LNP preparation hydrodynamic diameter and polydispersity index (PDI) is measured using high throughput dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt). LNP preparations are diluted 1X PBS to an appropriate concentration and analyzed. Concentration & Encapsulation Efficiency [0500] Concentration of NA is determined by Qubit microRNA kit (for siRNA) or HS RNA kit (for mRNA) per manufacturer’s instructions. Encapsulation efficiency is determined by measuring unlysed and lysed LNPs. LNP Administration and Base Editing [0501] Four weeks prior to dosing, NBSGW mice aged approximately 8-12 weeks are engrafted with 5e5 human CD34+ cells and used in the experiments described herein. Four weeks post- engraftment, each mouse is temporarily restrained, and LNP preparations are administered IV at 3 mg/kg via tail vein injection. [0502] Age-matched mice are also used to administer vehicle (1X PBS) via tail vein injection as a control. Four to six days post-dose, tissues including liver, spleen, and bone marrow are collected. Base editing is determined by performing targeted deep sequencing analysis at the human ALAS1 target site using extracted genomic DNA. Genomic DNA is isolated and fragmented and adapter-ligated using the Nextera DNA Flex Library Prep Kit (Illumina) using the 96-well plate Nextera indexing primers (Illumina), according to the manufacturer’s instructions. Library size and concentration is confirmed by Fragment Analyzer (Agilent) and is sequenced using whole genome sequencing with an Illumina HiSeq. Equivalents [0503] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims: