CORNEBISE MARK (US)
HENNESSY EDWARD (US)
KUMARASINGHE ELLALAHEWAGE S (US)
WO2017075531A1 | 2017-05-04 | |||
WO2017049245A2 | 2017-03-23 | |||
WO2019152557A1 | 2019-08-08 | |||
WO2018170306A1 | 2018-09-20 | |||
WO2017049275A2 | 2017-03-23 | |||
WO2017062513A1 | 2017-04-13 |
A. R. GENNARO: "Remington's The Science and Practice of Pharmacy", 2006, LIPPINCOTT, WILLIAMS & WILKINS
SMITH, M. B.MARCH, J.: "March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure,", 2001, JOHN WILEY & SONS
GREENE, T.W.WUTS, P.G. M.: "Protective Groups in Organic Synthesis", 1999, JOHN WILEY & SONS
R. LAROCK: "Comprehensive Organic Transformations", 1989, VCH PUBLISHERS
L. FIESERM. FIESER: "Fieser and Fieser's Reagents for Organic Synthesis", 1994, JOHN WILEY AND SONS
"Encyclopedia of Reagents for Organic Synthesis", 1995, JOHN WILEY AND SONS
CLAIMS 1. A compound of Formula (A): its N-oxide, or a salt or isomer thereof, wherein R’a is R’branched or R’cyclic; wherein wherein denotes a point of attachment; wherein Raa is H, and Rab, Rag, and Rad are each independently selected from the group consisting of H, C2-12 alkyl, and C2-12 alkenyl, wherein at least one of Rab, Rag, and Rad is selected from the group consisting of C2-12 alkyl and C2-12 alkenyl; R2 and R3 are each C1-14 alkyl; R4 is selected from the group consisting of -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH wherein R10 is N(R)2; each R is independently selected from the group consisting of C1-6 alkyl, C2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R5 is independently selected from the group consisting of OH, C1-3 alkyl, C2-3 alkenyl, and H; each R6 is independently selected from the group consisting of OH, C1-3 alkyl, C2-3 alkenyl, and H; R7 is H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C1-12 alkyl or C2-12 alkenyl; Ya is a C3-6 carbocycle; R*”a is selected from the group consisting of C1-15 alkyl and C2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. 2. A compound of Formula (B): r its N-oxide, or a salt or isomer thereof, wherein R’a is R’branched or R’cyclic; wherein wherein denotes a point of attachment; wherein Raa and Rab are each H, and Rag and Rad are each independently selected from the group consisting of H, C2-12 alkyl, and C2-12 alkenyl, wherein at least one of Rag and Rad is selected from the group consisting of C2-12 alkyl and C2-12 alkenyl; Rba, Rbb, Rbg, and Rbd are each independently selected from the group consisting of H, C2-30 alkyl, and C5-20 alkenyl, wherein at least one of Rba, Rbb, Rbg, and Rbd is selected from the group consisting of C2-30 alkyl and C5-20 alkenyl; R4 is selected from the group consisting of -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, - (CH2)5OH and , wherein denotes a point of attachment; R10 is N(R)2; each R is independently selected from the group consisting of C1-6 alkyl, C2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R5 is independently selected from the group consisting of OH, C1-3 alkyl, C2-3 alkenyl, and H; each R6 is independently selected from the group consisting of OH, C1-3 alkyl, C2-3 alkenyl, and H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C1-12 alkyl or C2-12 alkenyl; Ya is a C3-6 carbocycle; R*”a is selected from the group consisting of C1-15 alkyl and C2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. 3. A compound of Formula (1-1): or its N-oxide, or a salt or isomer thereof, wherein R’a is R’branched or R’cyclic; wherein wherein denotes a point of attachment; wherein Rag and Rbg are each independently a C2-12 alkyl or C2-12 alkenyl; R2 and R3 are each independently selected from the group consisting of C1-14 alkyl and C2-14 alkenyl; R4 is -(CH2)2OH; each R’ independently is a C1-12 alkyl or C2-12 alkenyl; Ya is a C3-6 carbocycle; R*”a is selected from the group consisting of C1-15 alkyl and C2-15 alkenyl; and s is 2 or 3. 4. The compound of any one of the preceding claims, having one of the following structures: c 5. The compound of any one of the preceding claims, wherein Rag is a C2-6 alkyl. 6. The compound of any one of the preceding claims, wherein Rbg is a C2-6 alkyl. 7. The compound of any one of the preceding claims, wherein Rag and Rbg are each independently a C2-6 alkyl. 8. The compound of any one of the preceding claims, wherein Rbg is a C4-6 alkyl. 9. The compound of any one of the preceding claims, wherein R2 and R3 are each C8 alkyl. 10. The compound of any one of the preceding claims, wherein Ya is cyclohexyl or cyclopentyl. 11. The compound of any one of the preceding claims, wherein R*”a is a C2-alkyl or C3- alkyl. 12. The compound of any one of the preceding claims, wherein each R’ independently is a C2-5 alkyl. 13. A compound selected from: 14. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding claims, a phospholipid, a structural lipid, and a PEG lipid. 15. The empty LNP of any one of the preceding claims, comprising about 40 mol % to about 60 mol % said compound, about 0 mol % to about 20 mol % phospholipid, about 30 mol % to about 50 mol % structural lipid, and about 0 mol % to about 5 mol % PEG lipid. 16. The empty LNP of any one of the preceding claims, wherein the phospholipid is selected from the group consisting of: 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine,1,2-dioleoyl-sn-glycero-3-phosphoetha nolamine (DOPE), 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), sphingomyelin, and mixtures thereof. 17. The empty LNP of any one of the preceding claims, wherein the structural lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, and mixtures thereof. 18. The empty LNP of any one of the preceding claims, wherein the PEG lipid is selected from the group consisting of a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. 19. The empty LNP of any one of the preceding claims, wherein the PEG lipid is selected from PEG2k-DMG and PEG-1: 20. A loaded lipid nanoparticle (loaded LNP), which comprises the empty LNP of any one of the preceding claims and one or more therapeutic and/or prophylactic agents. 21. The loaded LNP of any one of the preceding claims, wherein the one or more therapeutic and/or prophylactic agents is a nucleic acid. 22. The loaded LNP of any one of the preceding claims, wherein the nucleic acid is an RNA, and wherein the RNA is selected from the group consisting of a short interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a RNA interference (RNAi) molecule, a microRNA (miRNA), an antagomir, an antisense RNA, a ribozyme, a Dicer-substrate RNA (dsRNA), a small hairpin RNA (shRNA), a messenger RNA (mRNA), and mixtures thereof. 23. The loaded LNP of any one of the preceding claims, wherein the RNA is an mRNA. 24. A pharmaceutical composition comprising the loaded LNP of any one of the preceding claims and a pharmaceutically acceptable carrier. 25. A method of delivering a therapeutic and/or prophylactic agent to a cell within a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding claims. 26. A method of specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding claims. 27. A method of producing a polypeptide of interest in a cell within a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding claims. 28. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the loaded LNP of any one of the preceding claims. 29. The method of any one of the preceding claims, wherein the organ is selected from the group consisting of liver, kidney, lung, and spleen. 30. The method of any one of the preceding claims wherein the administering is performed parenterally, intramuscularly, intradermally, subcutaneously, and/or intravenously. |
[00326] As illustrated in Scheme 2 above, an acid a2 (x 3 is an integer between 1 and 7; e.g., 8-bromooctanoic acid) reacts with an alcohol b2 (e.g., nonan-1-ol) to afford an ester c2 (e.g., nonyl-8-bromooctanoate). Step 1 can take place in an organic solvent (e.g., dichloromethane) in the presence of, e.g., N-(3-dimethylaminopropyl)-N¢-ethylcarbodiimide hydrochloride, N,N-diisopropylethylamine and DMAP. Alcohol e2 (e.g., heptadecan-9-ol) can be obtained from reacting aldehyde d2 (e.g., nonanal) with a Grignard reagent R 3 -MgX (e.g., n-C8H17MgBr) via Step 2. Next, 8-bromooctanoic acid reacts with an alcohol e2 (e.g., heptadecan-9-ol) to afford an ester f2 (e.g., heptadecan-9-yl 8-bromooctanoate). Step 3 can take place in an organic solvent (e.g., dichloromethane) in the presence of, e.g., N-(3- dimethylaminopropyl)-N¢-ethylcarbodiimide hydrochloride, N,N-diisopropylethylamine and DMAP. Next, ester f2 reacts with 2-aminoethan-1-ol to afford amine g2 (e.g., heptadecan-9- yl 8-((2-hydroxyethyl)amino)octanoate). Step 4 can take place in ethanol in the presence of i-Pr2EtN. Then amine g2 reacts with ester c2 (e.g., nonyl-8-bromooctanoate) to afford compound h2 (e.g., heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8- oxooctyl)amino)octanoate). Step 5 can take place in an organic solvent (e.g., a mixture of CPME and MeCN), in the presence of a base (such as an inorganic base (e.g., K 2 CO 3 ) or non- nucleophilic organic base (e.g., i-Pr2EtN)) and a catalyst (e.g., an iodide such as KI or NaI) at, e.g., an elevated temperature (such as at about 70-90 ° C, e.g., about 80 ° C). [00327] As illustrated in Scheme 3 above, a haloalkanol (x 3 is an integer between 1 and 12, e.g., 6- bromohexan-1-ol) is reacted with a starting material a3 (x 2 is an integer between 1 and 6, e.g., 4-(hexyloxy)-4-oxobutanoic acid) to afford a halogenated diester b3 (e.g., 6- bromohexyl hexyl succinate). Compound a3 can be obtained by reaction of an alcohol (e.g., hexan-1-ol) with an acid anhydride (e.g. succinic anhydride, dihydro-2H-pyran-2,6(3H)- dione, 3-(tert-butoxy)-3-oxopropanoic acid, 4-(tert-butoxy)-3-methyl-4-oxobutanoic acid, or 4-(tert-butoxy)-2-methyl-4-oxobutanoic acid). Step 1 can take place in an organic solvent (e.g., dichloromethane) in the presence of, e.g., N-(3-dimethylaminopropyl)-N¢- ethylcarbodiimide hydrochloride, N,N-diisopropylethylamine and DMAP. Next, halogenated diester b3 reacts with an amine c3 (x 4 is an integer between 5 and 13, x 5 is an integer between 1 and 5, e.g., heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate) to afford the product d3. Step 2 can take place in an organic solvent (e.g., a mixture of CPME and MeCN), in the presence of a base (such as an inorganic base (e.g., K 2 CO 3 ) and a catalyst (e.g., an iodide such as KI) and an ether solvent (e.g., cyclopentyl methyl ether), at an elevated temperature (e.g., about 90 ° C). [00328] A person of ordinary skill in the art will recognize that in the above schemes the order of certain steps may be interchangeable. [00329] In certain aspects, the disclosure also includes methods of synthesizing a compound of any of Formulae (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B- 1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), and (B- c) and intermediate(s) for synthesizing the compound. [00330] In some embodiments, the method of synthesizing a compound of the disclosure includes reacting a compound of Formula (X2): with R 1 -Br to afford the compound of the disclosure, wherein each variables are as defined herein. For example, m is 5, 6, 7, 8, or 9, preferably 5, 7, or 9. For example, each of R 5 , R 6 , and R 7 is H. For example, M is -C(O)O- or -OC(O)-. For example, R 4 is unsubstituted C 1-3 alkyl, or -(CH 2 )nQ, in which n is 2, 3, or 4 and Q is OH, -NHC(S)N(R) 2 , -NHC(O)N(R) 2 , -N(R)C(O)R, or - N(R)S(O) 2 R. For example, the reaction of the compound of Formula (X2) with R 1 -Br takes place in the presence of a base (such as an inorganic base (e.g., K 2 CO 3 ) or non-nucleophilic organic base (e.g., i-Pr2EtN)). For example, the reaction takes place in the presence of an inorganic base (e.g., K 2 CO 3 ) and a catalyst (e.g., an iodide such as KI or NaI). For example, the reaction takes place at an elevated temperature, e.g., about 50-100 ° C, 70-90 ° C, or about 80 ° C). [00331] The method may also include reacting a compound of Formula (X1): with R 4 NH 2 to afford a compound of Formula (X2), wherein each variables are as defined herein. [00332] In some embodiments, the intermediate(s) include those having any of Formulae (X1) and (X2): wherein each variables are as defined herein. For example, the intermediate includes heptadecan-9-yl 8- bromooctanoate, and heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate, and morphic forms thereof (e.g., a crystalline form). [00333] In addition, it is to be understood that any particular embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Since such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. [00334] All cited sources, for example, references, publications, databases, database entries, and art cited herein, are incorporated into this application by reference, even if not expressly stated in the citation. In case of conflicting statements of a cited source and the instant application, the statement in the instant application shall control. Examples Example 1: Synthesis of compounds of Table 1 A. General Considerations [00335] All solvents and reagents used were obtained commercially and used as such unless noted otherwise. 1 H NMR spectra were recorded in CDCl 3 , at 300 K using a Bruker Ultrashield 300 MHz instrument. Chemical shifts are reported as parts per million (ppm) relative to TMS (0.00) for 1 H. Silica gel chromatographies were performed on ISCO CombiFlash Rf+ Lumen Instruments using ISCO RediSep Rf Gold Flash Cartridges (particle size: 20-40 microns). Reverse phase chromatographies were performed on ISCO CombiFlash Rf+ Lumen Instruments using RediSep Rf Gold C18 High Performance columns. All final compounds were determined to be greater than 85% pure via analysis by reverse phase UPLC-MS (retention times, RT, in minutes) using Waters Acquity UPLC instrument with DAD and ELSD and a ZORBAX Rapid Resolution High Definition (RRHD) SB-C18 LC column, 2.1 mm, 50 mm, 1.8 µm, and a gradient of 65 to 100% acetonitrile in water with 0.1% TFA over 5 minutes at 1.2 mL/min. Injection volume was 5 ^L and the column temperature was 80 °C. Detection was based on electrospray ionization (ESI) in positive mode using Waters SQD mass spectrometer (Milford, MA, USA) and evaporative light scattering detector. LCMS method: Instrument Information: HPLC/MS-Agilent 1100 Column: Agela Technologies Durashell C183.5 mm, 100 Å, 4.6 × 50 mm Mobile Phase A: Water/0.1% Trifluoroacetic Acid Mobile Phase B: Acetonitrile/0.1% Trifluoroacetic Acid Flow Rate: 1 mL/min Gradient: 70% B to 100% B in 5 minutes, hold 100% B for 10 minutes, 100% B to 70% B in minute, and then stop. Column Temperature: Ambient Detector: ELSD [00336] The procedures described below are useful in the synthesis of compounds of Table 1. [00337] The following abbreviations are employed herein: THF: Tetrahydrofuran MeCN: Acetonitrile LAH: Lithium Aluminum Hydride DCM: Dichloromethane DMAP: 4-Dimethylaminopyridine LDA: Lithium Diisopropylamide rt: Room Temperature DME: 1,2-Dimethoxyethane n-BuLi: n-Butyllithium CPME: Cyclopentyl methyl ether i-Pr2EtN: N,N-Diisopropylethylamine Representative Procedure A: 1,4-Addition of Grignard Reagent RMgX to Methyl (E)- non-2-enoate, 1 A1. Compound 2a: Methyl 3-butylnonanoate [00338] To an oven-dried 100 mL round bottom flask was added copper(I) bromide (421.3 mg, 2.93 mmol) and lithium chloride (249 mg, 5.87 mmol), then dry THF (15 mL) was added and the mixture was stirred for 10 min during which time the solids were dissolved. The flask was put in an ice bath and methyl (E)- non-2-enoate 1 (5 g, 29.37 mmol) was added, followed by addition of TMSCl 4 mL, 32.31 mmol). The reaction was stirred for 15 min. A THF solution of butylmagnesium bromide (17.6 mL, 35.2 mmol, 2.0 M in THF) was added slowly and the reaction was stirred for 2 h. The reaction was quenched with saturated NH 4 Cl (10 mL) and extracted with diethyl ether (100 mL) and dried over anhydrous sodium sulfate. After removing the solvent, the crude was purified by flash chromatography (SiO 2 : ethyl acetate/hexane 0-100%) and colorless oil product 2a was obtained (3 g, 45%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.64 (s, 3H); 2.21 (d, 2H, J = 6.9 Hz); 1.85-1.81 (m, 1H); 1.23-1.20 (m, 16H); 0.86-0.84 (m, 6H). A2. Compound 2b: Methyl 3-isopropylnonanoate [00339] Same as the procedure A1 but using isoproypylmagnesium bromide. Yield = 2.4 g (38%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.65 (s, 3H); 2.25 (dd, 1H, J = 15.1, 6.1 Hz); 2.14 (dd, 1H, J = 15.1, 7.1 Hz); 1.23-1.20 (m, 12H); 0.87-0.84 (m, 9H). A3. Compound 2c: Methyl 3-propylnonanoate [00340] Same as the procedure A1 but using n-proypylmagnesium bromide. Yield = 2.2 g (35%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.64 (s, 3H); 2.23 (d, 2H, J = 6.8 Hz); 1.85-1.84 (m, 1H); 1.23-1.20 (m, 14H); 0.87-0.84 (m, 6H). Representative Procedure B: LAH Reduction B1. Compound 3a: 3-Butylnonan-1-ol [00341] A solution of the methyl 3-butylnonanoate, 2a (2.2 g, 9.63 mmol) in THF (10 mL) was added dropwise to a stirred suspension of LiAlH4 (0.73 g, 19.27 mmol) in THF (10 mL) under N 2 . The mixture was heated under reflux for 5 h. The reaction was cooled to room temperature. Under 0 ℃ ice-water bath, 0.7 mL of H2O, 0.7 mL 15% NaOH, 2.1 mL of H2O were added sequentially. The white precipitate was filtered, and the filtrate was concentrated. The crude product was purified by flash chromatography (SiO2: ethyl acetate/hexane 0- 100%) and colorless oil product 3a was obtained (980 mg, 51%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.64 (t, 2H, J = 6.8 Hz); 1.52 (q, 2H, J = 7.1 Hz); 1.32-1.30 (m, 1H); 1.23- 1.20 (m, 17H); 0.88-0.84 (m, 6H). B2. Compound 3b: 3-Isopropylnonan-1-ol [00342] Same as the procedure B1 but using Methyl 3-isopropylnonanoate, 2b. Yield = 1.7 g (81%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.64 (m, 2H); 1.55 (m, 2H); 1.43-1.41 (m, 1H); 1.23-1.20 (m, 12H); 0.88-0.84 (m, 9H). B3. Compound 3c: 3-Propylnonan-1-ol [00343] Same as the procedure B1 but using Methyl 3-propylnonanoate, 2c. Yield = 1.28 g (67%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.64 (t, 2H, J = 6.6 Hz); 1.52 (q, 2H, J = 6.3 Hz); 1.23-1.20 (m, 16H); 0.88-0.84 (m, 6H). Representative Procedure C for Esterification of 8-Bromooctanoic Acid, 4 C1. Compound 5a: 3-Butylnonyl 8-bromooctanoate [00344] To a solution of 3-Butylnonan-1-ol 3a (458 mg, 2.28 mmol), 8-bromooctanoic acid 4 (611.9 mg, 2.74 mmol) and DMAP (55.9 mg, 0.46 mmol) in dichloromethane (30 mL) at 0 ˚C was added EDCI (657.3 mg, 3.43 mmol) and the reaction mixture stirred at room temperature overnight. TLC showed the completed reaction. The reaction mixture was cooled to 0 ˚C and 1N hydrochloric acid (3 mL) was added slowly, then the mixture was extracted with diethyl ether (100 mL) and the layers were separated. The organic layer washed with saturated sodium bicarbonate (100 mL), water and brine. The organic layer was separated and concentrated. The crude was purified by flash chromatography (SiO 2 : hexane/diethyl ether 0- 100%) and colorless oil product 5a was obtained (680 mg.73%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 4.07 (t, 2H, J = 6.8 Hz); 3.39 (t, 2H, J = 6.8 Hz); 2.28 (t, 2H, J = 7.6 Hz); 1.88-1.79 (m, 2H); 1.70-1.42 (m, 6H); 1.38-1.17 (m, 21H); 0.88-0.82 (m, 6H). C2. Compound 5b: 3-Isopropylnonyl 8-bromooctanoate [00345] Same as the procedure C1 but using 3-Isopropylnonan-1-ol, 3b. Yield = 297 mg (71%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 4.05 (dd, 2H, J = 14.3, 6.6 Hz); 3.39 (t, 2H, J = 6.8 Hz); 2.28 (t, 2H, J = 7.7 Hz); 1.86-1.81 (m, 2H); 1.70-1.42 (m, 6H); 1.38-1.17 (m, 16H); 0.84-0.82 (m, 9H). C3. Compound 5c: 3-Propylnonyl 8-bromooctanoate [00346] Same as the procedure C1 but using 3-Propylnonan-1-ol, 3c. Yield = 430 mg (68%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.96 (d, 2H, J = 5.8 Hz); 3.38 (t, 2H, J = 5.5 Hz); 2.27 (t, 2H, J = 7.4 Hz); 1.88-1.79 (m, 2H); 1.70-1.42 (m, 6H); 1.38-1.17 (m, 19H); 0.88-0.82 (m, 6H). Representative Procedure D: N-alkylation of Heptadecan-9-yl 8-((2- hydroxyethyl)amino)octanoate, 6 D1. Compound 7: 3-Butylnonyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(2- hydroxyethyl)amino)octanoate (86-g-nBu) [00347] In a 500 mL round bottom flask connected with condenser, heptadecan-9-yl 8-((2- hydroxyethyl)amino)octanoate 6 (601 mg, 1.36 mmol), 3-butylnonyl 8-bromooctanoate 5a (606 mg, 1.49 mmol), potassium carbonate (676 mg, 4.9 mmol) and potassium iodide (248.4 mg, 1.49 mmol) were mixed in cyclopentylmethyl ether (30 mL) and acetonitrile (30 mL), and the reaction mixture was heated to 85 °C for 18 h. MS showed clean conversion, and the mixture was cooled to room temperature and diluted with hexanes. The mixture was filtered through pad of Celite. After washing with hexanes, the filtrate was concentrated to give brown oil which was purified by flash chromatography (SiO2: hexane/diethyl ether 0-100%) to afford 7 as a colorless oil (588 mg.56%). HPLC/ELSD: RT = 7.07 min. MS (CI): m/z (MH + ) 766.7 for C 4 8H95NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.85 (quint., 1H, J = 6.1 Hz); 4.07 (t, 2H, J = 6.9 Hz); 3.50 (t, 2H, J = 5.5 Hz); 2.98 (bs, 1H); 2.55 (t, 2H, J = 5.2 Hz); 2.41 (t, 4H, J = 7.4 Hz); 2.26 (t, 4H, J = 7.4 Hz); 1.65-1.48 (m, 19H); 1.26 (br. m, 48H); 0.88-0.84 (m, 12H). D2. Compound 12: Heptadecan-9-yl 8-((2-hydroxyethyl)(8-((3-isopropylnonyl)oxy)-8- oxooctyl)amino)octanoate (86-g-iPr) [00348] Same as the procedure D1 but using 3-Isopropylnonyl 8-bromooctanoate, 5b. Yield = 258 mg (50%). HPLC/ELSD: RT = 6.98 min. MS (CI): m/z (MH + ) 752.6 for C 4 7H93NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.85 (quint., 1H, J = 6.1 Hz); 4.07 (m, 2H); 3.50 (t, 2H, J = 5.2 Hz); 3.01 (bs, 1H); 2.55 (t, 2H, J = 5.2 Hz); 2.41 (t, 4H, J = 7.4 Hz); 2.26 (dd, 4H, J = 7.6, 2.7 Hz); 1.65-1.48 (m, 14H); 1.26 (br. m, 48H); 0.88-0.84 (m, 15H). D3. Compound 13: Heptadecan-9-yl 8-((2-hydroxyethyl)(8-oxo-8-((3- propylnonyl)oxy)octyl)amino)octanoate (86-g-nPr) [00349] Same as the procedure D1 but using 3-Propylnonyl 8-bromooctanoat, 5c. Yield = 510 mg (68%). HPLC/ELSD: RT = 7.01 min. MS (CI): m/z (MH + ) 752.6 for C 47 H 93 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.85 (quint., 1H, J = 6.3 Hz); 4.07 (t, 2H, J = 7.1 Hz); 3.50 (t, 2H, J = 5.5 Hz); 2.98 (bs, 1H); 2.55 (t, 2H, J = 5.2 Hz); 2.41 (t, 4H, J = 7.4 Hz); 2.26 (t, 4H, J = 7.4 Hz); 1.65-1.48 (m, 17H); 1.26 (br. m, 48H); 0.88-0.84 (m, 12H). Synthetic Scheme for Preparation of Compound 8 C4. Compound 5d: 2-Propylnonyl 8-bromooctanoate [00350] Same as procedure C1 but using 2-propylnonan-1-ol 15a. Yield = 1.67 g (79%). 1 H NMR (300 MHz, CDCl 3 ): d ppm 3.96 (d, 2H, J = 5.8 Hz); 3.38 (t, 2H, J = 5.5 Hz); 2.27 (t, 2H, J = 7.4 Hz); 1.88-1.79 (m, 2H); 1.70-1.42 (m, 6H); 1.38-1.17 (m, 19H); 0.88-0.82 (m, 6H). D4. Compound 8: Heptadecan-9-yl 8-((2-hydroxyethyl)(8-oxo-8-((2- propylnonyl)oxy)octyl)amino)octanoate (86-b-nPr) [00351] Same as procedure D1 but using 2-Propylnonyl 8-bromooctanoate, 5d. Yield = 355 mg (68%). HPLC/ELSD: RT = 7.0 min. MS (CI): m/z (MH + ) 752.6 for C 47 H 93 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.85 (quint., 1H, J = 6.3 Hz); 3.95 (d, 2H, J = 5.8 Hz); 3.50 (t, 2H, J = 5.5 Hz); 3.02 (bs, 1H); 2.55 (t, 2H, J = 5.5 Hz); 2.41 (t, 4H, J = 7.7 Hz); 2.26 (dd, 4H, J = 13.9, 6.6 Hz); 1.65-1.48 (m, 17H); 1.26 (br. m, 48H); 0.88-0.84 (m, 12H). Synthesis of Intermediates: Intermediate AA: Ethyl 3-propylhex-2-enoate [00352] Triethyl phosphonoacetate (11.3 mL, 56.9 mmol) was added dropwise over 20 minutes to a suspension of sodium hydride (2.28 g, 56.9 mmol) in THF (17 mL) and the mixture was stirred at room temperature until gas evolution ceased (approximately 30 min). The reaction mixture was chilled to 0 °C and 4-heptanone (6.12 mL, 43.8 mmol) was added in portions. The reaction was gradually warmed to room temperature and allowed to stir under reflux for 24 h. The reaction was cooled to room temperature prior to being quenched with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with diethyl ether, and the organic extracts were washed with brine, dried (MgSO 4 ), and concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford ethyl 3-propylhex-2-enoate (8.07 g, 43.8 mmol, 100%) as a clear oil and as a mixture of regioisomers. 1 H NMR (300 MHz, CDCl 3 ) as a mixture of regioisomers d: ppm 5.63 (s, 1H); 5.38-5.25 (m, 0.74H); 4.19-4.07 (m, 3.40H); 3.02 (s, 0.81H); 2.96 (s, 0.59H); 2.57 (ddd, 2H, J = 6.0, 6.0, 3.0 Hz); 2.16-1.98 (m, 4.87H); 1.57-1.35 (m, 6.10H); 1.34-1.21 (m, 7.59H); 1.01-0.82 (m, 12.9H). Intermediate AB: Ethyl 3-propylhexanoate [00353] A steel Parr reactor equipped with a stir bar was charged with ethyl 3-propylhex- 2-enoate (8.07 g, 43.8 mmol) in ethanol (44 mL). Palladium hydroxide on carbon (922 mg, 6.57 mmol) was added and the vessel was sealed, evacuated, refilled with H 2 gas (3x), and the pressure was set to 200 psi. The reaction was stirred at 500 rpm, under 200 psi H2 gas, at room temperature for 2 h. The vessel was then evacuated, refilled with N 2 gas, and opened. The crude reaction mixture was filtered through a Celite pad. The Celite pad was washed with EtOH and the crude material was concentrated to give ethyl 3-propylhexanoate (6.55 g, 35.2 mmol, 80%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.12 (q, 2H, J = 6.0 Hz); 2.22 (d, 2H, J = 9.0 Hz); 1.95-1.81 (m, 1H); 1.38-1.18 (m, 11H); 0.89 (br. t, 6H, J = 6.0 Hz). Intermediate AC: 3-Propylhexan-1-ol [00354] To a mixture of lithium aluminum hydride (1.60 g, 42.2 mmol) in dry ether (42 mL) under N 2 at 0°C, was added dropwise ethyl 3-propylhexanoate (6.55 g, 35.2 mmol) in dry ether (28 mL). The mixture was stirred at room temperature for 2.5 h prior to being cooled to 0° C. Water (1 mL per g of LiAlH 4 ) was added to the solution dropwise, followed by the slow addition of 15% sodium hydroxide (1 mL per g of LiAlH4) and water (3 mL per g of LiAlH 4 ). The solution was stirred for a few minutes at room temperature and filtered through a Celite pad. The Celite pad was washed with diethyl ether and the filtrate was concentrated. The crude material was purified by silica gel chromatography (0-40% EtOAc:hexanes) to afford 3-propylhexan-1-ol (4.82 g, 33.4 mmol, 95%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.67 (t, 2H, J = 6.0 Hz); 1.57-1.39 (m, 3H); 1.37-1.18 (m, 9H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AD: Ethyl 3-butylhept-2-enoate [00355] Triethyl phosphonoacetate (9.07 mL, 45.7 mmol) was added dropwise over 20 minutes to a suspension of sodium hydride (1.83 g, 45.7 mmol) in THF (14 mL) and the mixture was stirred at room temperature until gas evolution ceased (approximately 30 min). The reaction mixture was chilled to 0 °C and 5-nonanone (6.05 mL, 35.2 mmol) was added in portions. The reaction was gradually warmed to room temperature and allowed to stir under reflux for 24 h. The reaction was cooled to room temperature prior to being quenched with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with diethyl ether, and the organic extracts were washed with brine, dried (MgSO 4 ), and concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford ethyl 3- butylhept-2-enoate (5.27 g, 24.8 mmol, 71%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.62 (s, 1H); 4.14 (q, 2H, J = 6.0 Hz); 2.59 (t, 2H, J = 6.0 Hz); 2.14 (t, 2H, J = 6.0 Hz); 1.50-1.23 (m, 11H); 0.99-0.82 (m, 6H). Intermediate AE: Ethyl 3-butylheptanoate [00356] A steel Parr reactor equipped with a stir bar was charged with ethyl 3-butylhept-2- enoate (10.5 g, 49.5 mmol) in ethanol (50 mL). Palladium hydroxide on carbon (1.04 g, 7.42 mmol) was added and the vessel was sealed, evacuated, refilled with H2 gas (3x), and the pressure was set to 200 psi. The reaction was stirred at 500 rpm, under 200 psi H2 gas, at room temperature for 2 h. The vessel was then evacuated, refilled with N 2 gas, and opened. The crude reaction mixture was filtered through a Celite pad. The Celite pad was washed with EtOH and the crude material was concentrated to give ethyl 3-butylheptanoate (9.69 g, 45.2 mmol, 91%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.12 (q, 2H, J = 9.0 Hz); 2.22 (d, 2H, J = 6.0 Hz); 1.90-1.76 (m, 1H); 1.38-1.19 (m, 15H); 0.88 (br. t, 6H, J = 6.0 Hz). Intermediate AF: 3-Butylheptan-1-ol [00357] To a mixture of lithium aluminum hydride (850 mg, 22.4 mmol) in dry ether (23 mL) under N 2 at 0°C, was added dropwise ethyl 3-butylheptanoate (4.00 g, 18.7 mmol) in dry ether (15 mL). The mixture was stirred at room temperature for 2.5 h prior to being cooled to 0° C. Water (1 mL per g of LiAlH 4 ) was added to the solution dropwise, followed by the slow addition of 15% sodium hydroxide (1 mL per g of LiAlH4) and water (3 mL per g of LiAlH 4 ). The solution was stirred for a few minutes at room temperature and filtered through a Celite pad. The Celite pad was washed with diethyl ether and the filtrate was concentrated. The crude material was purified by silica gel chromatography (0-40% EtOAc:hexanes) to afford 3-butylheptan-1-ol (3.19 g, 18.5 mmol, 99%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.66 (t, 2H, J = 6.0 Hz); 1.53 (q, 2H, J = 6.0 Hz); 1.46-1.36 (m, 1H); 1.35- 1.21 (m, 12H); 1.18 (br. s, 1H); 0.89 (br. t, 6H, J = 6.0 Hz). Intermediate AG: Ethyl 3-pentyloct-2-enoate [00358] Triethyl phosphonoacetate (10.6 mL, 53.4 mmol) was added dropwise over 20 minutes to a suspension of sodium hydride (2.13 g, 53.4 mmol) in THF (16 mL) and the mixture was stirred at room temperature until gas evolution ceased (approximately 30 min). The reaction mixture was chilled to 0 °C and 6-undecanone (8.42 mL, 41.1 mmol) was added in portions. The reaction was gradually warmed to room temperature and allowed to stir under reflux for 60 h. The reaction was cooled to room temperature prior to being quenched with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with diethyl ether, and the organic extracts were washed with brine, dried (MgSO 4 ), and concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford ethyl 3-pentyloct-2-enoate (8.76 g, 36.5 mmol, 89%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.61 (s, 1H); 4.14 (q, 2H, J = 6.0 Hz); 2.58 (ddd, 2H, J = 9.0, 9.0, 6.0 Hz); 2.13 (ddd, 2H, J = 6.0, 6.0, 3.0 Hz); 1.52-1.38 (m, 3H); 1.38-1.23 (m, 12H); 0.93-0.86 (m, 6H). Intermediate AH: Ethyl 3-pentyloctanoate
[00359] A steel Parr reactor equipped with a stir bar was charged with ethyl 3-pentyloct-2- enoate (8.76 g, 36.5 mmol) in ethanol (37 mL). Palladium hydroxide on carbon (768 mg, 5.47 mmol) was added and the vessel was sealed, evacuated, refilled with H2 gas (3x), and the pressure was set to 200 psi. The reaction was stirred at 500 rpm, under 200 psi H 2 gas, at room temperature for 2 h. The vessel was then evacuated, refilled with N2 gas, and opened. The crude reaction mixture was filtered through a Celite pad. The Celite pad was washed with EtOH and the crude material was concentrated to give ethyl 3-pentyloctanoate (8.45 g, 34.9 mmol, 96%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.12 (q, 2H, J = 6.0 Hz); 2.22 (d, 2H, J = 6.0 Hz); 1.92-1.77 (br. m, 1H); 1.37-1.19 (m, 19H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AI: 3-Pentyloctan-1-ol [00360] To a mixture of lithium aluminum hydride (1.59 g, 41.8 mmol) in dry ether (42 mL) under N 2 at 0°C, was added dropwise ethyl 3-pentyloctanoate (8.45 g, 34.9 mmol) in dry ether (28 mL). The mixture was stirred at room temperature for 2.5 h prior to being cooled to 0° C. Water (1 mL per g of LiAlH 4 ) was added to the solution dropwise, followed by the slow addition of 15% sodium hydroxide (1 mL per g of LiAlH4) and water (3 mL per g of LiAlH 4 ). The solution was stirred for a few minutes at room temperature and filtered through a Celite pad. The Celite pad was washed with diethyl ether and the filtrate was concentrated. The crude material was purified by silica gel chromatography (0-40% EtOAc:hexanes) to afford 3-pentyloctan-1-ol (6.98 g, 34.9 mmol, 100%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.66 (t, 2H, J = 6.0 Hz); 1.53 (q, 2H, J = 6.0 Hz); 1.47-1.37 (br. s, 1H); 1.36- 1.15 (m, 17H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AJ: 3-Pentyloctanal [00361] To a stirred suspension of pyridinium chlorochromate (9.02 g, 41.8 mmol) and silica gel (9.02 g, 1g/g of pyridinium chlorochromate) in dichloromethane (90 mL) under a N2 atmosphere was added 3-pentyloctan-1-ol (6.98 g, 34.9 mmol). The suspension was stirred at room temperature for 1 h. The reaction was then filtered through a Celite pad, the Celite pad was washed with dichloromethane, and the filtrate was concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford 3-pentyloctanal (4.66 g, 23.5 mmol, 67%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 9.76 (t, 1H, J = 3.0 Hz); 2.33 (dd, 2H, J = 6.0, 3.0 Hz); 2.01-1.86 (br. m, 1H); 1.40-1.19 (m, 16H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AK: 6-Allylundecane [00362] To a suspension of methyltriphenylphosphonium bromide (4.68 g, 13.1 mmol) in dry ether (190 mL) under a N 2 was added potassium tert-butoxide (1.47 g, 13.1 mmol) in one portion. The mixture was stirred at room temperature for 15 minutes, prior to the dropwise addition of 3-pentyloctanal (2.00 g, 10.1 mmol) in dry ether (26 mL) over 15 min. The resulting mixture was allowed to stir for 90 min at room temperature. The reaction mixture was diluted with ice water, the layers were separated, and the organic layer was extracted with ether. The combined organics were dried (MgSO 4 ), filtered, and concentrated. The crude material was purified by silica gel chromatography (0-10% EtOAc:hexanes) to afford 6- allylundecane (1.65 g, 8.38 mmol, 83%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.77 (dddd, 1H, J = 15.0, 12.0, 9.0, 9.0 Hz); 5.03-4.94 (m, 2H); 2.02 (ddd, 2H, J = 9.0, 6.0, 6.0 Hz); 1.43-1.16 (m, 17H); 0.88 (d, 6H, J = 6.0 Hz). Intermediate AL: 4-Pentylnonan-1-ol [00363] To a stirred solution of sodium borohydride (131 mg, 3.46 mmol) in dry diglyme (3.6 mL) under a N 2 atmosphere was added a solution of 6-allylundecane (2.26 g, 11.5 mmol) in dry diglyme (2.3 mL). Next, a solution of boron trifluoride etherate (569 mL, 4.61 mmol) in 1.2 mL of dry diglyme was added over 15 min at room temperature. The resulting mixture was stirred for 1 hour prior to the dropwise addition of water (1.2 mL). When gas evolution ceased, 2.3 mL of 3M NaOH was added at room temperature, followed by the dropwise addition of 2.3 mL of 30% H2O2 at 40 °C. After 1 hour of stirring at 40 °C, the reaction was poured into 10 mL of water. The reaction vessel was washed with additional water. The combined water solutions were extracted with ether (2x). Combined ethereal extracts were washed with water (x5). Ethereal extracts were dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-40% EtOAc:hexanes) to afford 4-pentylnonan-1-ol (1.88 g, 8.77 mmol, 76%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.62 (t, 2H, J = 6.0 Hz); 1.60-1.48 (m, 2H); 1.37-1.19 (m, 20H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AM: 3-Propylhexyl 8-bromooctanoate [00364] To a solution of 3-propylhexan-1-ol (4.82 g, 33.4 mmol), 8-bromooctanoic acid (8.94 g, 40.1 mmol), and DMAP (816 mg, 6.68 mmol) in methylene chloride (58 mL) at 0 ˚C was added EDCI (9.60 g, 50.1 mmol) and the reaction mixture stirred at room temperature overnight. The reaction mixture was then cooled to 0 ˚C and a solution of 10% hydrochloric acid (180 mL) was added slowly over 20 minutes. The layers were separated, and the organic layer was concentrated in vacuum to give a crude oil. The oil was dissolved in hexane (180 mL) and washed with a mixture of acetonitrile (180 mL) and 5% sodium bicarbonate (180 mL). The hexane layer was separated, dried (MgSO 4 ), and filtered. The solvent was removed under vacuum to give 3-propylhexyl 8-bromooctanoate (10.9 g, 31.2 mmol, 93%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 2H, J = 6.0 Hz); 3.40 (t, 2H, J = 6.0 Hz); 2.28 (t, 2H, J = 6.0 Hz); 1.85 (pent., 2H, J = 6.0 Hz); 1.68-1.51 (m, 4H); 1.49-1.18 (m, 15H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AN: 3-Butylheptyl 8-bromooctanoate [00365] To a solution of 3-butylheptan-1-ol (3.19 g, 18.5 mmol), 8-bromooctanoic acid (4.96 g, 22.2 mmol), and DMAP (453 mg, 3.71 mmol) in methylene chloride (32 mL) at 0 ˚C was added EDCI (5.33 g, 27.8 mmol) and the reaction mixture stirred at room temperature overnight. The reaction mixture was then cooled to 0 ˚C and a solution of 10% hydrochloric acid (150 mL) was added slowly over 20 minutes. The layers were separated, and the organic layer was concentrated in vacuum to give a crude oil. The oil was dissolved in hexane (150 mL) and washed with a mixture of acetonitrile (150 mL) and 5% sodium bicarbonate (150 mL). The hexane layer was separated, dried (MgSO 4 ), and filtered. The solvent was removed under vacuum to give 3-butylheptyl 8-bromooctanoate (6.90 g, 18.3 mmol, 99%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 2H, J = 6.0 Hz); 3.40 (t, 2H, J = 6.0 Hz); 2.29 (t, 2H, J = 6.0 Hz); 1.85 (pent., 2H, J = 6.0 Hz); 1.69-1.52 (m, 4H); 1.49-1.20 (m, 19H); 0.89 (br. t, 6H, J = 6.0 Hz). Intermediate AO: 3-Pentyloctyl 8-bromooctanoate [00366] To a solution of 3-pentyloctan-1-ol (2.00 g, 9.98 mmol), 8-bromooctanoic acid (2.67 g, 12.0 mmol), and DMAP (244 mg, 2.00 mmol) in methylene chloride (18 mL) at 0 ˚C was added EDCI (2.87 g, 15.0 mmol) and the reaction mixture stirred at room temperature overnight. The reaction mixture was then cooled to 0 ˚C and a solution of 10% hydrochloric acid (70 mL) was added slowly over 20 minutes. The layers were separated, and the organic layer was concentrated in vacuum to give a crude oil. The oil was dissolved in hexane (70 mL) and washed with a mixture of acetonitrile (70 mL) and 5% sodium bicarbonate (70 mL). The hexane layer was separated, dried (MgSO 4 ), and filtered. The solvent was removed under vacuum to give 3-pentyloctyl 8-bromooctanoate (3.94 g, 9.72 mmol, 97%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 2H, J = 6.0 Hz); 3.40 (t, 2H, J = 6.0 Hz); 3.29 (t, 2H, J = 6.0 Hz); 1.85 (pent., 2H, J = 6.0 Hz); 1.68-1.52 (m, 4H); 1.49-1.19 (m, 23H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AP: 4-Pentylnonyl-8-bromooctanoate [00367] To a solution of 4-pentylnonan-1-ol (1.88 g, 8.77 mmol), 8-bromooctanoic acid (2.35 g, 10.5 mmol), and DMAP (214 mg, 1.75 mmol) in methylene chloride (15 mL) at 0 ˚C was added EDCI (2.52 g, 13.2 mmol) and the reaction mixture stirred at room temperature overnight. The reaction mixture was then cooled to 0 ˚C and a solution of 10% hydrochloric acid (60 mL) was added slowly over 20 minutes. The layers were separated, and the organic layer was concentrated in vacuum to give a crude oil. The oil was dissolved in hexane (60 mL) and washed with a mixture of acetonitrile (60 mL) and 5% sodium bicarbonate (60 mL). The hexane layer was separated, dried (MgSO 4 ), and filtered. The solvent was removed under vacuum to give 4-pentylnonyl-8-bromooctanoate (3.68 g, 8.77 mmol, 100%) as a clear oil. The compound was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.04 (t, 2H, J = 6.0 Hz); 3.40 (t, 2H, J = 6.0 Hz); 2.29 (t, 2H, J = 6.0 Hz); 1.85 (pent., 2H, J = 6.0 Hz); 1.70-1.52 (m, 4H); 1.50-1.18 (m, 25H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AQ: Pentadecan-8-yl 8-bromooctanoate [00368] To a solution of 8-bromooctanoic acid (1.98 g, 8.87 mmol) in dichloromethane (30 mL) were added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.13 g, 11.1 mmol), 4-(dimethylamino)pyridine (0.217 g, 1.77 mmol), and pentadecan-8-ol (2.03 g, 8.87 mmol). The reaction was allowed to stir at room temperature for 18 hours. The reaction mixture was cooled to 0 ˚C and a solution of 10% hydrochloric acid was added slowly. The organic layer was separated and evaporated under vacuum. The residue was dissolved in hexanes and washed with a 1:1 mixture of acetonitrile and saturated NaHCO 3 (aq.). The hexane layer was separated, died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to obtain a mixture of ~13.2:1 pentadecan-8-yl 8-bromooctanoate and pentadecan-8-yl 8- chlorooctanoate (3.19 g, 83.1%) as a colorless liquid. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 3.55 (t, 0.14H); 3.42 (t, 1.86H); 2.31 (t, 2H); 1.88 (p, 2H); 1.72-1.59 (m, 2H); 1.59-1.42 (m, 6H); 1.42-1.18 (m, 24H); 0.90 (t, 6H). Intermediate AR: Tridecan-7-yl 8-bromooctanoate [00369] To a solution of 8-bromooctanoic acid (1.96 g, 8.76 mmol) in dichloromethane (30 mL) were added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.10 g, 10.9 mmol), 4-(dimethylamino)pyridine (0.234 g, 1.92 mmol), and tridecan-7-ol (1.75 g, 8.73 mmol). The reaction was allowed to stir at room temperature for 18 hours. The reaction was diluted with dichloromethane and extracted with saturated NaHCO 3 (aq.). The organic layer was separated and washed with brine, died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to obtain a mixture of ~12.3:1 tridecan-7-yl 8-bromooctanoate and tridecan-7-yl 8- chlorooctanoate (2.10 g, 59.4%) as a colorless liquid. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 3.55 (t, 0.15H); 3.42 (t, 1.85H); 2.31 (t, 2H); 1.88 (p, 2H); 1.72-1.60 (m, 2H); 1.60-1.42 (m, 6H); 1.42-1.19 (m, 20H); 0.90 (t, 6H). Intermediate AS: Undecan-6-yl 8-bromooctanoate [00370] To a solution of 8-bromooctanoic acid (4.00 g, 17.9 mmol) in dichloromethane (60 mL) were added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (4.31 g, 22.5 mmol), 4-(dimethylamino)pyridine (0.438 g, 3.58 mmol), and 6-undecanol (3.09 g, 17.9 mmol). The reaction was allowed to stir at room temperature for 18 hours The reaction mixture was cooled to 0 ˚C and a solution of 10% hydrochloric acid was added slowly. The organic layer was separated and evaporated under vacuum. The residue was dissolved in hexanes and washed with a 1:1 mixture of acetonitrile and saturated NaHCO 3 (aq.). The hexane layer was separated, died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to obtain a mixture of ~ 19:1 undecan-6-yl 8-bromooctanoate and undecan-6-yl 8- chlorooctanoate (4.33 g, 64.01%) as a colorless liquid. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 3.55 (t, 0.10H); 3.42 (t, 1.90H); 2.31 (t, 2H); 1.88 (p, 2H); 1.72-1.59 (m, 2H); 1.59-1.42 (m, 6H); 1.42-1.18 (m, 16H); 0.90 (t, 6H). Intermediate AT: Nonan-5-yl 8-bromooctanoate Molecular Weight: 349.35 [00371] To a solution of 8-bromooctanoic acid (4.00 g, 17.9 mmol) in dichloromethane (60 mL) were added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (4.31 g, 22.5 mmol), 4-(dimethylamino)pyridine (0.438 g, 3.59 mmol), and 5-nonanol (2.59 g, 17.9 mmol). The reaction was allowed to stir at room temperature for 18 hours. The reaction mixture was cooled to 0 ˚C and a solution of 10% hydrochloric acid was added slowly. The organic layer was separated and evaporated under vacuum. The residue was dissolved in hexanes and washed with a 1:1 mixture of acetonitrile and saturated NaHCO 3 (aq.). The hexane layer was separated, died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography (0-10% ethyl acetate in hexanes) to obtain a mixture of ~7:1 nonan-5-yl 8-bromooctanoate and nonan-5-yl 8-chlorooctanoate (5.23 g, 83.5%) as a colorless liquid. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.90 (p, 1H); 3.55 (t, 0.25H); 3.42 (t, 1.75H); 2.31 (t, 2H); 1.88 (p, 2H); 1.72-1.59 (m, 2H); 1.59-1.19 (m, 18H); 0.91 (t, 6H). Intermediate AU: 3-Propylhexyl 8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate [00372] To a solution of tert-butyl N-(3-aminopropyl)carbamate (23.9 g, 137 mmol) in EtOH (60 mL) was added 3-propylhexyl 8-bromooctanoate (8.00 g, 22.3 mmol) in EtOH (55 mL) over the course of 20 min. The reaction was heated to 60 °C and allowed to stir at this temperature for 16 h. Upon cooling, the solvents were evaporated and the residue was diluted with ethyl acetate and washed with saturated aqueous NaHCO 3 and brine (5X) until no white precipitate was observed in the aqueous layer. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The residue was purified by flash chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-propylhexyl 8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate (5.81 g, 13.1 mmol, 57%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.16 (br. s, 1H); 4.08 (t, 2H, J = 6.0 Hz); 3.19 (br. q, 2H, J = 6.0 Hz); 2.65 (t, 2H, J = 6.0 Hz); 2.56 (t, 2H, J = 6.0 Hz); 2.27 (t, 2H, J = 6.0 Hz); 1.70-1.51 (m, 6H); 1.50-1.39 (m, 3H); 1.43 (s, 9H); 1.36-1.17 (m, 15H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AV: 3-Propylhexyl 8-((2-hydroxyethyl)amino)octanoate [00373] To a round bottom flask equipped with a stir bar was added 3-propylhexyl 8- bromooctanoate (2.82 g, 8.06 mmol), ethanolamine (14.6 mL, 242 mmol), and ethyl alcohol (6 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-propylhexyl 8-((2- hydroxyethyl)amino)octanoate (876 mg, 2.66 mmol, 33%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 2H, J = 6.0 Hz); 3.63 (t, 2H, J = 6.0 Hz); 2.77 (t, 2H, J = 6.0 Hz); 2.61 (t, 2H, J = 6.0 Hz); 2.28 (t, 2H, J = 6.0 Hz); 1.91 (br. s, 2H); 1.68-1.39 (m, 7H); 1.38- 1.18 (m, 14H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AW: 3-Pentyloctyl 8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate [00374] To a solution of tert-butyl N-(3-aminopropyl)carbamate (15.5 g, 88.8 mmol) in EtOH (38 mL) was added 3-pentyloctyl 8-bromooctanoate (6.00 g, 14.8 mmol) in EtOH (36 mL) over the course of 20 min. The reaction was heated to 60 °C and allowed to stir at this temperature for 16 h. Upon cooling, the solvents were evaporated and the residue was diluted with ethyl acetate and washed with saturated aqueous NaHCO3 and brine (5X) until no white precipitate was observed in the aqueous layer. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The residue was purified by flash chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate (4.23 g, 8.49 mmol, 57%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.17 (br. s, 1H); 4.07 (t, 2H, J = 6.0 Hz); 3.19 (br. q, 2H, J = 6.0 Hz); 2.66 (t, 2H, J = 6.0 Hz); 2.56 (t, 2H, J = 6.0 Hz); 2.28 (t, 2H, J = 6.0 Hz); 1.70-1.52 (m, 6H); 1.51-1.39 (m, 3H); 1.44 (s, 9H); 1.36-1.19 (m, 22H); 0.88 (t, 6H, J = 6.0 Hz). Intermediate AX: 4-Pentylnonyl 8-((2-hydroxyethyl)amino)octanoate [00375] To a round bottom flask equipped with a stir bar was added 4-pentylnonyl 8- bromooctanoate (600 mg, 1.43 mmol), ethanolamine (2.59 mL, 42.9 mmol), and ethyl alcohol (1 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 4-pentylnonyl 8-((2- hydroxyethyl)amino)octanoate (306 mg, 0.77 mmol, 54%) as a clear oil. UPLC/ELSD: RT = 1.66 min. MS (ES): m/z (MH + ) 400.31 for C 24 H 49 NO 3 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.97 (t, 2H, J = 6.0 Hz); 3.57 (br. t, 2H, J = 6.0 Hz); 2.81 (br. s, 2H), 2.67 (br. t, 2H, J = 6.0 Hz); 2.53 (t, 2H, J = 6.0 Hz); 2.22 (t, 2H, J = 6.0 Hz); 1.61-1.35 (m, 6H); 1.32-1.10 (m, 25H); 0.81 (t, 6H, J = 6.0 Hz). Intermediate AY: 3-Pentyloctyl 8-((3-hydroxypropyl)amino)octanoate [00376] To a round bottom flask equipped with a stir bar was added 3-pentyloctyl 8- bromooctanoate (1.00 g, 2.47 mmol), propanolamine (5.66 mL, 74.0 mmol), and ethyl alcohol (2 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((3- hydroxypropyl)amino)octanoate (374 mg, 0.94 mmol, 38%) as a clear oil. UPLC/ELSD: RT = 1.64 min. MS (ES): m/z (MH + ) 400.18 for C 24 H 49 NO 3 . Intermediate AZ: Heptadecan-9-yl 8-((3-hydroxypropyl)amino)octanoate [00377] To a round bottom flask equipped with a stir bar was added heptadecane-9-yl 8- bromooctanoate (1.00 g, 2.17 mmol), propanolamine (4.97 mL, 65.0 mmol), and ethyl alcohol (2 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-((3- hydroxypropyl)amino)octanoate (723 mg, 1.59 mmol, 73%) as a clear oil. UPLC/ELSD: RT = 2.06 min. MS (ES): m/z (MH + ) 456.17 for C 2 8H57NO3. Intermediate BA: Heptadecan-9-yl 8-((4-hydroxybutyl)amino)octanoate [00378] To a round bottom flask equipped with a stir bar was added heptadecane-9-yl 8- bromooctanoate (1.00 g, 2.17 mmol), 4-aminobutan-1-ol (5.99 mL, 65.0 mmol), and ethyl alcohol (2 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-((4- hydroxybutyl)amino)octanoate (773 mg, 1.65 mmol, 76%) as a clear oil. UPLC/ELSD: RT = 2.02 min. MS (ES): m/z (MH + ) 470.23 for C 2 9H59NO3. Intermediate BB: 3-Pentyloctyl 8-((4-hydroxybutyl)amino)octanoate [00379] To a round bottom flask equipped with a stir bar was added 3-pentyloctyl 8- bromooctanoate (1.00 g, 2.47 mmol), 4-aminobutan-1-ol (6.82 mL, 74.0 mmol), and ethyl alcohol (2 mL). The resulting mixture was allowed to stir at 40 °C for 16h. The reaction was diluted with dichloromethane, washed with water (2x), and the layers were separated. The organic layer was dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((4- hydroxybutyl)amino)octanoate (501 mg, 1.21 mmol, 49%) as a clear oil. UPLC/ELSD: RT = 1.67 min. MS (ES): m/z (MH + ) 414.24 for C 25 H 51 NO 3 . Intermediate BC: 3-Pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00380] To a solution of 3-propylhexyl 8-bromooctanoate (735 mg, 2.11 mmol) and 3- pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (1.00 g, 2.01 mmol) in cyclopentyl methyl ether (9 mL) and actonitrile (9 mL) was added potassium carbonate (1.66 g, 12.0 mmol) and iodopotassium (366 mg, 2.21 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling, the volatiles were evaporated under vacuum. The residue was diluted with dichloromethane and washed with water. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((3-((tert- butoxycarbonyl)amino)propyl)(8-oxo-8-((3-propylhexyl)oxy)oct yl)amino)octanoate (698 mg, 0.91 mmol, 45%) as a golden oil. UPLC/ELSD: RT = 2.82 min. MS (ES): m/z (MH + ) 767.59 for C 46 H 90 N 2 O 6 . Intermediate BD: 3-Butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00381] To a solution of 3-butylheptyl 8-bromooctanoate (895 mg, 2.37 mmol) and 3- propylhexyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (1.00 g, 2.26 mmol) in cyclopentyl methyl ether (10 mL) and actonitrile (10 mL) was added potassium carbonate (1.87 g, 13.6 mmol) and iodopotassium (412 mg, 2.49 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling, the volatiles were evaporated under vacuum. The residue was diluted with dichloromethane and washed with water. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (972 mg, 1.32 mmol, 58%) as a golden oil. UPLC/ELSD: RT = 2.69 min. MS (ES): m/z (MH + ) 739.46 for C 4 4H86N2O6. Intermediate BE: Bis(3-propylhexyl) 8,8'-((3-((tert- butoxycarbonyl)amino)propyl)azanediyl)dioctanoate [00382] To a solution of 3-propylhexyl 8-bromooctanoate (829 mg, 2.37 mmol) and 3- propylhexyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (1.00 g, 2.26 mmol) in cyclopentyl methyl ether (10 mL) and actonitrile (10 mL) was added potassium carbonate (1.87 g, 13.6 mmol) and iodopotassium (412 mg, 2.49 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling, the volatiles were evaporated under vacuum. The residue was diluted with dichloromethane and washed with water. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-propylhexyl) 8,8'-((3-((tert -butoxycarbonyl)amino)propyl)azanediyl)dioctanoate (730 mg, 1.03 mmol, 45%) as a clear viscous oil. UPLC/ELSD: RT = 2.58 min. MS (ES): m/z (MH + ) 711.59 for C 42 H 82 N 2 O 6 . Intermediate BF: 3-Butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate [00383] To a solution of 3-butylheptyl 8-bromooctanoate (794 mg, 2.11 mmol) and 3- pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (1.00 g, 2.01 mmol) in cyclopentyl methyl ether (9 mL) and actonitrile (9 mL) was added potassium carbonate (1.66 g, 12.0 mmol) and iodopotassium (366 mg, 2.21 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling, the volatiles were evaporated under vacuum. The residue was diluted with dichloromethane and washed with water. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-((tert- butoxycarbonyl)amino)propyl)(8-oxo-8-((3-pentyloctyl)oxy)oct yl)amino)octanoate (896 mg, 1.13 mmol, 56%) as a clear oil. UPLC/ELSD: RT = 2.95 min. MS (ES): m/z (MH + ) 795.59 for C 4 8H94N2O6. Intermediate BG: 3-Pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8- (undecan-6-yloxy)octyl)amino)octanoate Molecular Weight: 795.288 [00384] UPLC/ELSD: RT= 2.93 min. MS (ESI): m/z calcd for C 48 H 95 N 2 O 6 + (M+H) 795.288; found, 795.71. 1H NMR (300 MHz, CDCl 3 ) d: ppm 5.66 (br. s, 1H); 4.88 (p, 1 H); 4.09 (t, 2H); 3.18 (br. d, 2H); 2.50 (br. d, 2H); 2.32 (br. d, 3H); 2.29 (t, 4H); 1.65-1.46 (m, 28H); 1.27 (m, 44H); 0.90 (t, 12H). Intermediate BH: Nonan-5-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate [00385] To a solution of nonan-5-yl 8-bromooctanoate (882 mg, 2.53 mmol) and 3- pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (1.20 g, 2.41 mmol) in cyclopentyl methyl ether (11 mL) and actonitrile (11 mL) was added potassium carbonate (2.00 g, 14.4 mmol) and iodopotassium (439 mg, 2.65 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling, the volatiles were evaporated under vacuum. The residue was diluted with dichloromethane and washed with water. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give nonan-5-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate (1.06 g, 1.39 mmol, 58%) as a golden oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.65 (br. s, 1H); 4.87 (pent., 2H, J = 6.0 Hz); 4.08 (t, 2H, J = 6.0 Hz); 3.18 (br. q, 2H, J = 6.0 Hz); 2.44 (br. s, 2H); 2.35 (br. s, 2H); 2.28 (t, 4H, J = 6.0 Hz); 1.71-1.17 (m, 53H); 1.43 (s, 9H); 0.88 (t, 12H, J = 6.0 Hz). Intermediate BI: Pentadecan-8-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8- ((3-propylhexyl)oxy)octyl)amino)octanoate Chemical Formula: C 4 8H94N2O6 Molecular Weight: 795.288 [00386] UPLC/ELSD: RT= 2.68 min. MS (ESI): m/z calcd for C 48 H 95 N 2 O 6 + (M+H) 795.288; found, 795.71. 1H NMR (300 MHz, CDCl 3 ) d: ppm 5.66 (br. s, 1H); 4.89 (p, 1 H); 4.10 (t, 2H); 3.19 (br. d, 2H); 2.56-2.35 (br. d, 5H); 2.30 (t, 5H); 1.66-1.39 (m, 66H); 0.90 (t, 12H). Intermediate BJ: 3-Propylhexyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8- (tridecan-7-yloxy)octyl)amino)octanoate [00387] To a solution of 3-propylhexyl 8-({3-[(tert- butoxycarbonyl)amino]propyl}amino)octanoate (1.38 g, 3.11 mmol) in Acetonitrile (9 mL) were added potassium iodide (0.588 g, 3.54 mmol), potassium carbonate (1.73 g, 12.5 mmol), and a solution of tridecan-7-yl 8-bromooctanoate (1.26 g, 3.11 mmol) in CMPE (9 mL). The reaction was allowed to stir at 77 °C for 18 hours. The reaction was cooled to room temperature and filtered, then the filtrate was evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3-propylhexyl 8-({3-[(tert- butoxycarbonyl)amino]propyl}[8-oxo-8-(tridecan-7-yloxy)octyl ]amino)octanoate (1.32 g, 55.1%) as a yellow tinted oil. UPLC/ELSD: RT = 2.70 min found, 767.34. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.67 (br. s, 1H); 4.89 (p, 1H); 4.10 (t, 2H); 3.20 (q, 2H); 2.61-2.43 (m, 2H); 2.43-2.35 (m, 4H); 2.30 (dt, 4H); 1.71-1.49 (m, 14H); 1.49-1.40 (m, 12H); 1.40-1.19 (m, 36H); 1.01-0.83 (m, 12H). Intermediate BK: 3-Propylhexyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8- (undecan-6-yloxy)octyl)amino)octanoate [00388] To a solution of 3-propylhexyl 8-({3-[(tert- butoxycarbonyl)amino]propyl}amino)octanoate (1.50 g, 3.39 mmol) in Acetonitrile (10 mL) were added potassium iodide (0.619 g, 3.73 mmol), potassium carbonate (1.87 g, 13.6 mmol), and a solution of undecan-6-yl 8-bromooctanoate (1.28 g, 3.39 mmol) in CPME (10 mL). The reaction was allowed to stir at 77 °C for 18 hours. The reaction was cooled to room temperature and filtered, then the filtrate was evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3-propylhexyl 8-({3-[(tert- butoxycarbonyl)amino]propyl}[8-oxo-8-(undecan-6-yloxy)octyl] amino)octanoate (1.53 g, 61.2%) as a yellow tinted oil. UPLC/ELSD: RT = 2.56 min found, 739.46. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.66 (br. s, 1H); 4.89 (p, 1H); 4.10 (t, 2H); 3.20 (q, 2H); 2.60-2.44 (m, 2H); 2.44-2.35 (m, 4H); 2.30 (t, 4H); 1.74-1.49 (m, 14H); 1.49-1.39 (m, 12H); 1.39-1.19 (m, 32H); 0.91 (t, 12H). Intermediate BL: Nonan-5-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00389] To a solution of 3-propylhexyl 8-({3-[(tert- butoxycarbonyl)amino]propyl}amino)octanoate (1.50 g, 3.39 mmol) in acetonitrile (10 mL) were added potassium iodide (0.619 g, 3.73 mmol), potassium carbonate (1.87 g, 13.6 mmol), and a solution of nonan-5-yl 8-bromooctanoate (1.18 g, 3.39 mmol) in CMPE (10 mL). The reaction was allowed to stir at 77 °C for 18 hours. The reaction was cooled to room temperature and filtered, then the filtrate was evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain nonan-5-yl 8-({3-[(tert- butoxycarbonyl)amino]propyl}({8-oxo-8-[(3-propylhexyl)oxy]oc tyl})amino)octanoate (0.483 g, 20.1%) as a yellow tinted oil. UPLC/ELSD: RT = 2.45 min found, 711.46. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.66 (br. s, 1H); 4.89 (p, 1H); 4.11 (t, 2H); 3.28-3.11 (m, 2H); 2.60-2.44 (m, 2H); 2.44-2.35 (m, 4H); 2.30 (t, 4H); 1.74-1.49 (m, 14H); 1.49-1.39 (m, 12H); 1.39-1.20 (m, 28H); 0.91 (t, 12H). Intermediate BM: 3-Pentyloctyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00390] To a solution of 3-pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo- 8-((3-propylhexyl)oxy)octyl)amino)octanoate (698 mg, 0.91 mmol) in methylene chloride (18 mL) was added trifluoroacetic acid (1.39 mL, 18.2 mmol). The reaction was allowed to stir at room temperature for 4 h. The reaction was quenched with saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (378 mg, 0.57 mmol, 62%) as a clear oil. UPLC/ELSD: RT = 2.26 min. MS (ES): m/z (MH + ) 667.56 for C 4 1H82N2O4. Intermediate BN: 3-Butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00391] To a solution of 3-butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo- 8-((3-propylhexyl)oxy)octyl)amino)octanoate (972 mg, 1.32 mmol) in methylene chloride (27 mL) was added trifluoroacetic acid (2.01 mL, 26.3 mmol). The reaction was allowed to stir at room temperature for 4 h. The reaction was quenched with saturated aqueous NaHCO 3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (503 mg, 0.79 mmol, 60%) as a clear oil. UPLC/ELSD: RT = 2.13 min. MS (ES): m/z (MH + ) 639.31 for C 39 H 78 N 2 O 4 . Intermediate BO: Bis(3-propylhexyl) 8,8'-((3-aminopropyl)azanediyl)dioctanoate [00392] To a solution of bis(3-propylhexyl) 8,8'-((3-((tert - butoxycarbonyl)amino)propyl)azanediyl)dioctanoate (730 mg, 1.03 mmol) in methylene chloride (21 mL) was added trifluoroacetic acid (1.57 mL, 20.5 mmol). The reaction was allowed to stir at room temperature for 4 h. The reaction was quenched with saturated aqueous NaHCO 3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-propylhexyl) 8,8'-((3- aminopropyl)azanediyl)dioctanoate (499 mg, 0.82 mmol, 80%) as a clear oil. UPLC/ELSD: RT = 1.93 min. MS (ES): m/z (MH + ) 611.44 for C 37 H 74 N 2 O 4 . Intermediate BP: 3-Butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate [00393] To a solution of 3-butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo- 8-((3-pentyloctyl)oxy)octyl)amino)octanoate (896 mg, 1.13 mmol) in methylene chloride (23 mL) was added trifluoroacetic acid (1.72 mL, 22.5 mmol). The reaction was allowed to stir at room temperature for 4 h. The reaction was quenched with saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate (632 mg, 0.91 mmol, 81%) as a clear oil. UPLC/ELSD: RT = 2.47 min. MS (ES): m/z (MH + ) 695.68 for C 43 H 86 N 2 O 4. Intermediate BQ: 3-Pentyloctyl 8-((4-aminobutyl)(8-oxo-8-(undecan-6- yloxy)octyl)amino)octanoate [00394] UPLC/ELSD: RT= 2.49 min. MS (ESI): m/z calcd for C 4 4H89N2O6 + (M+H) 709.198; found, 695.43. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.09 (t, 2H); 2.74 (t, 2H); 2.47 (t, 2H); 2.39 (t, 4H); 2.29 (t, 4H); 1.69-1.38 (m, 25H); 1.29 (br. m, 38H); 0.90 (t, 12H). Intermediate BR: Nonan-5-yl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate [00395] To a solution of nonan-5-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-oxo-8- ((3-pentyloctyl)oxy)octyl)amino)octanoate (1.06 g, 1.39 mmol) in methylene chloride (28 mL) was added trifluoroacetic acid (2.12 mL, 27.7 mmol). The reaction was allowed to stir at room temperature for 4 h. The reaction was quenched with saturated aqueous NaHCO3 and extracted with dichloromethane. The organic layer was separated, washed with brine, dried (MgSO 4 ), filtered and concentrated. The crude material was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give nonan-5-yl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate (718 mg, 1.08 mmol, 78%) as a clear oil. UPLC/ELSD: RT = 2.31 min. MS (ES): m/z (MH + ) 667.43 for C 41 H 82 N 2 O 4. Intermediate BS: 3-Pentyloctyl 8-((3-aminopropyl)(8-oxo-8-(undecan-6- yloxy)octyl)amino)octanoate [00396] UPLC/ELSD: RT= 2.34 min. MS (ESI): m/z calcd for C 4 3H87N3O6 + (M+H) 695.171; found, 695.430. 1H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 4.10 (t, 2H); 2.92 (t, 2H); 2.61 (t, 2H); 2.46 (t, 4H); 2.30 (t, 4H); 1.73-1.41 (m, 18H); 1.28 (br. m, 40H); 0.90 (t, 12H). Intermediate BT: 3-Propylhexyl 8-((3-aminopropyl)(8-oxo-8-(tridecan-7- yloxy)octyl)amino)octanoate Chemical Formula: C 4 1H82N2O4 Molecular Weight: 667.12 [00397] To a solution of 3-propylhexyl 8-({3-[(tert-butoxycarbonyl)amino]propyl}[8-oxo- 8-(tridecan-7-yloxy)octyl]amino)octanoate (1.32 g, 1.72 mmol) in dichloromethane (33 mL) was added trifluoroacetic acid (6.63 mL, 34.3 mmol). The reaction was allowed to stir at room temperature for 4 hours. Saturated NaHCO 3 (aq.) was added and the reaction was diluted with dichloromethane. The organic layer was separated and washed twice more with saturated NaHCO 3 (aq.) then brine. The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3- propylhexyl 8-[(3-aminopropyl)[8-oxo-8-(tridecan-7-yloxy)octyl]amino]oct anoate (0.774 g, 67.6%) as a yellow tinted oil. UPLC/ELSD: RT = 2.03 min found, 667.31. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 2.79 (t, 2H); 2.51 (t, 2H); 2.42 (t, 4H); 2.30 (t, 4H); 1.79-1.39 (m, 17H); 1.39-1.18 (m, 36H); 0.90 (t, 12H). Intermediate BU: 3-Propylhexyl 8-((3-aminopropyl)(8-oxo-8-(undecan-6- yloxy)octyl)amino)octanoate [00398] To a solution of 3-propylhexyl 8-({3-[(tert-butoxycarbonyl)amino]propyl}[8-oxo- 8-(undecan-6-yloxy)octyl]amino)octanoate (1.53 g, 2.07 mmol) in dichloromethane (40 mL) was added trifluoroacetic acid (3.05 mL, 39.9 mmol). The reaction was allowed to stir at room temperature for 4 hours. Saturated NaHCO 3 (aq.) was added and the reaction was diluted with dichloromethane. The organic layer was separated and washed twice more with saturated NaHCO 3 (aq.) then brine. The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3- propylhexyl 8-[(3-aminopropyl)[8-oxo-8-(undecan-6-yloxy)octyl]amino]octa noate (0.682 g, 51.5%) as a yellow tinted oil. UPLC/ELSD: RT = 1.85 min found, 639.19. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.10 (t, 2H); 2.82 (t, 2H); 2.53 (t, 2H); 2.44 (t, 4H); 2.30 (t, 4H); 1.75-1.40 (m, 17H); 1.40-1.18 (m, 32H); 0.91 (t, 12H). Intermediate BV: Nonan-5-yl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00399] To a solution of nonan-5-yl 8-({3-[(tert-butoxycarbonyl)amino]propyl}({8-oxo-8- [(3-propylhexyl)oxy]octyl})amino)octanoate (0.483 g, 0.679 mmol) in dichloromethane (13 mL) was added trifluoroacetic acid (1.00 mL, 13.1 mmol). The reaction was allowed to stir at room temperature for 4 hours. Saturated NaHCO3 (aq.) was added and the reaction was diluted with dichloromethane. The organic layer was separated and washed twice more with saturated NaHCO 3 (aq.) then brine. The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain nonan-5-yl 8-[(3-aminopropyl)({8-oxo-8-[(3-propylhexyl)oxy]octyl})amino ]octanoate (0.202 g, 48.7%) as a yellow tinted oil. UPLC/ELSD: RT = 1.85 min found, 611.44. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.10 (t, 2H); 2.80 (t, 2H); 2.52 (t, 2H); 2.42 (t, 4H); 2.30 (t, 4H); 1.74-1.39 (m, 17H); 1.39-1.20 (m, 28H); 0.91 (t, 12H). Synthesis of Final Compounds: AA. Compound 3: Heptadecan-9-yl 8-((2-hydroxyethyl)(6-((((3- pentyloctyl)oxy)carbonyl)oxy)hexyl) amino)octanoate [00400] UPLC/ELSD: RT = 3.04 min. MS (ES): m/z (MH + ) 769.313 for C 47 H 93 NO 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.14 (m, 4H); 3.54 (bm, 2H); 2.66-2.37 (m, 6H); 2.30 (m, 2H); 1.77-1.17 (m, 66H); 0.91 (m, 12H). AB. Compound 4: Heptadecan-9-yl 8-((6-((((3-hexylnonyl)oxy)carbonyl)oxy)hexyl)(3- ((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)a mino)octanoate Ethyl 3-hexylnon-2-enoate [00401] Triethyl phosphonoacetate (26.33 g, 117.4 mmol) was added dropwise over 20 minutes to a suspension of sodium hydride (4.697 g, 117.4 mmol) in THF (294 mL) and the mixture was stirred at room temperature until gas evolution ceased (approximately 30 min). The reaction mixture was chilled to 0°C and 7-tridecanone (10 g, 58.7 mmol) was added. The reaction was gradually warmed to room temperature, then heated to reflux and stirred overnight. The reaction was quenched with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with diethyl ether, and the organic extracts were washed with brine, dried with MgSO 4 , and concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford ethyl 3-hexylnon-2-enoate (6.7 g, 27.9 mmol, 47.5%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.63 (s, 1H); 4.15 (q, 2H); 2.61 (t, 2H); 2.15 (t, 2H); 1.53-1.20 (m, 19H); 0.91 (m, 6H). Ethyl 3-hexylnonanoate [00402] To a flask containing a slurry of Pearlmans catalyst (0.73 g, 5.2 mmol) in ethanol (20 mL) under N 2 was added a solution of ethyl 3-hexylnon-2-enoate (6.975 g, 25.9 mmol) in ethanol (5 mL). The reaction was stirred under H2 (balloon) for 16h. The reaction was filtered through a plug of Celite and the filtrate was evaporated under vacuum to afford ethyl 3-hexylnonanoate (6.7 g, 24.7 mmol, 95 %). The residue was taken to the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.16 (q, 2H); 2.23 (d, 2H); 1.86 (bs, 1H); 1.28 (m, 23H); 0.90 (m, 6H). 3-Hexylnonan-1-ol [00403] To a solution of lithium aluminium hydride (49.5 mL of 1M solution in THF, 49.5 mmol) in THF was added a solution of ethyl 3-hexylnonanoate (6.7 g, 24.7 mmol) in THF (20 mL). The reaction was stirred at room temperature for 16h. The reaction was quenched with a saturated solution of sodium sulfate decahydrate. The white solids were removed by filtration through a plug of Celite and the filtrate was evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0- 100% ethyl acetate in hexanes to obtain 3-hexylnonan-1-ol (5.62 g, 24.6 mmol, 99%). 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.69 (t, 2H); 1.61-1.19 (m, 24H); 0.91 (m, 6H). 6-Bromohexyl 3-hexylnonyl carbonate [00404] 3-Hexylnonan-1-ol (1.24 g, 5.4 mmol) was added dropwise to a solution of 6- bromohexyl 4-nitrophenyl carbonate (1.88 g, 5.43 mmol) in DCM (15 mL) in a round bottom flask charged with a magnetic stir bar at room temperature under N2. The reaction was kept under N2 and pyridine (0.55 mL, 6.8 mmol) was added dropwise over 10 min. followed by 4- dimethylaminopyridine (0.133 g, 1.1 mmol) in one portion. The reaction was allowed to stir at room temperature for 16h then diluted with water and DCM. The organic layer was separated, and the aqueous layer was washed with DCM. The combined organics were washed with brine, dried with Na 2 SO 4 . and evaporated under vac. The residue was purified by silica gel chromatography (0-100% ethyl acetate in hexanes) to give 6-bromohexyl 3- hexylnonyl carbonate (1.3 g, 3.0 mmol, 55%). Heptadecan-9-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate [00405] A solution of heptadecan-9-yl 8-bromooctanoate (69.2 g, 0.15 mole) and tert- butyl (3-aminopropyl)carbamate (130.6 g, 0.75 mole) in 500 mL ethanol was heated to 65 °C overnight. The reaction mixture was concentrated, and the crude was purified by flash column chromatography (SiO2: methanol/dichloromethane 0-20%) to get heptadecan-9-yl 8- ((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (62 g, 74%) as light yellow oil. MS (CI): m/z (MH + ) 555.5 for C 3 3H66N2O4. 1 H NMR (300 MHz, CDCl 3 ): d ppm 5.15 (bs, 1H); 4.85 (quint., 1H, J = 6.0 Hz); 3.17 (m, 2H); 2.65 (t, 2H, J = 6.6 Hz); 2.56 (t, 2H, J = 6.8 Hz); 2.26 (t, 2H, J = 7.6 Hz); 1.68-1.56 (m, 6H); 1.46 (m, 5H); 1.43 (s, 9H); 1.24 (m, 30H); 0.86 (t, 6H, J = 6.6 Hz). Heptadecan-9-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(6-((((3- hexylnonyl)oxy)carbonyl)oxy)hexyl)amino)octanoate [00406] To a solution of 6-bromohexyl 3-hexylnonyl carbonate (0.5 g, 1.15 mmol) and heptadecan-9-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (0.637 g, 1.15 mmol) in 6 mL of a 1:1 mixture of cyclopropyl methyl ether and acetonitrile was added potassium carbonate (0.635 g, 4.59 mmol) and potassium iodide (0.21 g, 1.26 mmol). The reaction was allowed to stir at 77 o C for 16 h. The reaction was cooled, filtered and the volatiles were evaporated under vacuum. The residue was purified by silica gel chromatography (0-100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM) to give heptadecan-9-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)(6-((((3- hexylnonyl)oxy)carbonyl)oxy)hexyl)amino)octanoate (0.32 g, 0.35 mmol, 31%). Heptadecan-9-yl 8-((6-((((3-hexylnonyl)oxy)carbonyl)oxy)hexyl)(3-((2-(methyl amino)- 3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate [00407] Compound 4 was prepared analogously to compound 14 using heptadecan-9-yl 8- ((3-((tert-butoxycarbonyl)amino)propyl)(6-((((3- hexylnonyl)oxy)carbonyl)oxy)hexyl)amino)octanoate instead of undecan-3-yl 8-((3-(((tert- butoxycarbonyl)amino)propyl)(6-((2-octyldecyl)oxy)-6-oxohexy l)amino)octanoate. UPLC/ELSD: RT = 3.13 min. MS (ES): m/z (MH + ) 919.429 for C 55 H 103 N 3 O 7. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (p, 1H); 4.17 (m, 4H); 3.68 (bm, 2H); 3.30 (m, 3H); 2.65-2.41 (m, 6H); 2.31 (m, 2H); 1.87-1.19 (m, 73H); 0.90 (m, 12H). AC. Compound 5: Heptadecan-9-yl 8-((6-((((3-hexylnonyl)oxy)carbonyl)oxy)hexyl)(2- hydroxyethyl)amino)octanoate [00408] UPLC/ELSD: RT = 3.16 min. MS (ES): m/z (MH + ) 797.683 for C 49 H 97 NO 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.14 (m, 4H); 3.54 (bm, 2H); 2.68-2.38 (m, 6H); 2.30 (m, 2H); 1.77-1.17 (m, 70H), 0.90 (m, 12H). AD. Compound 6: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(6-((((3-pentyloctyl)oxy)carbonyl)oxy)hexyl) amino)octanoate [00409] Compound 6 was prepared analogously to compound 4 using 6-undecanone instead of 7-tridecanone. UPLC/ELSD: RT = 3.04 min. MS (ES): m/z (MH + ) 891.552 for C 5 3H99N3O7. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (p, 1H); 4.17 (m, 4H); 3.67 (bm, 2H); 3.28 (m, 3H); 2.68-2.38 (m, 6H); 2.31 (m, 2H); 1.86-1.18 (m, 69H); 0.90 (m, 12H). AE. Compound 9: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl) amino)propyl)(8-oxo-8-((2-propylnonyl)oxy)octyl)amino)octano ate Heptadecan-9-yl 8-((3-(((benzyloxy)carbonyl)amino)propyl)amino) octanoate [00410] To a solution of heptadecan-9-yl 8-bromooctanoate (50 g, 204 mmol) and benzyl (3-aminopropyl)carbamate (35 g, 76 mmol) in 500 mL ethanol was added sodium bicarbonate (57 g, 0.68 mole) in one portion at room temperature, the mixture heated to 65 o C and stirred for two days. The reaction mixture was cooled to room temperature and the solid was filtered away through a pad of Celite. The filtrate was concentrated and purified by column chromatography (dichloromethane/methanol 9:1) to give heptadecan-9-yl 8-((3- (((benzyloxy)carbonyl)amino)propyl)amino) octanoate (29.2 g, 66%) as a light yellow oil. 1 H NMR (300 MHz, CDCl 3 ): d 0.84 (m, 6H); 1.10-1.57 (m, 42H); 2.24 (t, 2H, J = 6.7 Hz); 2.49 (m, 2H); 2.56 (m, 2H); 3.04 (m, 2H); 4.76 (m, 1H); 4.99 (s, 2H);7.29-7.35 (m, 5H). tert-Butyl 8-((3-(((benzyloxy)carbonyl)amino)propyl)(8-(heptadecan-9-yl oxy)-8- oxooctyl)amino)octanoate [00411] To a solution of heptadecan-9-yl 8-((3- (((benzyloxy)carbonyl)amino)propyl)amino) octanoate (14.6 g, 24.9 mmol) in 500 mL cyclopentylmethyl ether/acetonitrile (1:1, v/v) at room temperature was added tert-butyl 8- bromooctanoate (Oakwood Chemical, Estill, SC; 7.62 g, 387 mL, 27.3 mmol), followed by potassium carbonate (13.7 g, 99.6 mmol) and potassium iodide (5 g, 30 mmol). The reaction mixture was stirred at room temperature for 30 min and then at 85 o C overnight. The reaction mixture was cooled to room temperature and the solids were removed through a pad of Celite. The filtrate was concentrated and purified by column chromatography (hexane/ethyl acetate, 9:1 to 1:1) to give tert-butyl 8-((3-(((benzyloxy)carbonyl)amino)propyl)(8- (heptadecan-9-yloxy)-8-oxooctyl)amino)octanoate (14.3 g, 73%) as a colorless oil. 1 H NMR (300 MHz, CDCl 3 ): d 0.84 (m, 6H); 1.10-1.64 (m, 59H); 2.17-2.32 (m, 8H); 2.42 (m, 2H); 3.26 (m, 2H); 4.84 (m, 1H); 5.07 (s, 2H); 6.20 (m, 1H); 7.29-7.35 (m, 5H). tert-Butyl 8-((3-aminopropyl)(8-(heptadecan-9-yloxy)-8-oxooctyl)amino) octanoate [00412] To a solution of tert-butyl 8-((3-(((benzyloxy)carbonyl)amino)propyl)(8- (heptadecan-9-yloxy)-8-oxooctyl)amino)octanoate (28.6 g, 36.3 mmol) in 500 mL ethanol was added palladium on carbon (3 g, 10% wet, matrix activated). The reaction mixture was stirred under a hydrogen balloon overnight. MS showed no more starting material, and the mixture was filtered through a pad of Celite. The filtrate was concentrated to give tert-butyl 8-((3-aminopropyl)(8-(heptadecan-9-yloxy)-8-oxooctyl)amino) octanoate (23.4 g, quant.) as a brown oil, which was used in the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ): d 0.84 (m, 6H); 1.22-1.71 (m, 61H); 2.18 (t, 2H, J = 6.7 Hz); 2.25 (t, 2H, J = 6.7Hz); 2.32 (m, 4H); 2.39 (t, 2H, J = 6.8 Hz); 2.70 (t, 2H, J = 6.7 Hz); 4.86 (m, 1H). tert-Butyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate [00413] To a solution of tert-butyl 8-((3-aminopropyl)(8-(heptadecan-9-yloxy)-8- oxooctyl)amino) octanoate (23.47 g, 36 mmol) in 500 mL diethyl ether at 0 ˚C, was added 3,4-dimethoxy cyclobut-3-ene-1,2-dione (5.63 g, 40 mmol) and the reaction mixture stirred at room temperature for 4 hours. Methylamine solution (2 M in methanol, 23.4 mL, 46.8 mmol) was added, and the reaction mixture stirred at room temperature overnight. The reaction mixture was concentrated, and the residue was triturated with 100 mL tetrahydrofuran. The solid was removed through a pad of Celite. The filtrate was concentrated and purified by column chromatography with dichloromethane to dichloromethane/methanol/NH 4 OH (9:1:0.1) to give tert-butyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate (23 g, 86%) as a white wax. HPLC/UV (254 nm, Method-B): RT = 6.73 min. MS (CI): m/z (MH + ) 762.5 for C 45 H 83 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ): d 0.84 (m, 6H); 1.22-1.64 (m, 57H); 1.78 (m, 2H); 2.18 (t, 2H, J = 6.7 Hz); 2.26 (t, 2H, J = 6.7 Hz); 8-((8-(Heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4-dioxocyclobut-1-en-1- yl)amino)propyl)amino)octanoic acid [00414] To a solution of tert-butyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino )octanoate (450 mg, 0.59 mmol) in DCM (9.7 mL) was added trifluoroacetic acid (2.4 mL, 32.0 mmol) at 0 °C. The resulting mixture was allowed to stir at room temperature for 4 h. The reaction mixture was then concentrated in vacuo and the crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoic acid (389 mg, 0.55 mmol, 93%) as a golden oil. UPLC/ELSD: RT = 2.12 min. MS (ES): m/z (MH + ) 706.41 for C 4 1H75N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 11.42 (br. s, 1H); 9.29 (br. s, 1H); 8.64 (br. s, 1H); 4.83 (pent., 1H, J = 6 Hz); 3.71 (br. t, 2H, J = 6 Hz); 3.27 (br. s, 3H); 3.14 (br. t, 2H, J = 6 Hz); 2.94 (br. t, 4H, J = 6 Hz); 2.31-2.14 (m, 4H); 2.04 (br. s, 2H); 1.77-1.10 (m, 48H); 0.85 (t, 6H, J = 6 Hz). 4-Methyleneundecane [00415] To a suspension of methyltriphenylphosphonium bromide (11 g, 31 mmol) in tetrahydrofuran at -78 ˚C, was added n-BuLi solution (2.5 M in hexane, 16.8 mL, 40 mmol) dropwise. The reaction mixture was slowly warmed up to 0 o C for 3 hours until all the solid was dissolved. A solution of undecan-4-one (5g, 29.5 mmol) in tetrahydrofuran was added, and then the reaction mixture was heated to reflux overnight. After the reaction mixture was cooled to room temperature, the solvent was removed under vacuum. The residue was purified by silica gel chromatography (pentane) to give 4-methyleneundecane (4.9 g, 99%) as colorless oil. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m, 12H); 1.97 (m, 4H); 4.68 (s, 2H). 2-Propylnonan-1-ol [00416] To a solution of 4-methyleneundecane (4.9 g, 29.2 mmol) in tetrahydrofuran was added borane- tetrahydrofuran complex (1 M in THF, 36 mL) dropwise at 0 ˚C. The reaction mixture was stirred at room temperature for 3 hours until TLC indicated no more starting material, and aqueous 4 M NaOH (40 mL) was added. After stirring for 10 min, 30% hydrogen peroxide (10 mL) was added and stirred for 4 hours. The reaction mixture was quenched by aqueous sodium bisulfite solution and extracted with ethyl acetate (50 mL x 3). The combined organic layer was dried (Na2SO4), concentrated and purified by silica gel chromatography (hexane/ethyl acetate) to give 2-propylnonan-1-ol (4.2g, 77%) as a colorless oil. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m, 17H); 3.52 (d, 2H, J = 5.5 Hz). Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl) amino)propyl)(8- oxo-8-((2-propylnonyl)oxy)octyl)amino)octanoate [00417] To a solution of 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoic acid (830 mg, 1.17 mmol) in dichloromethane was added 2-propylnonan-1-ol (500 mg, 2.7 mmol), EDCI (1.3 g, 6.75 mmol) and 4-dimethylaminopyridine (33 mg, 0.27 mmol), and the mixture was stirred at room temperature overnight. The reaction was quenched with water and the organic layer was separated. The aqueous layer was extracted with dichloromethane (50 mL x 3). The combined organics were dried (Na 2 SO 4 ), concentrated and purified by silica gel chromatography )dichloromethane/methanol/NH 4 OH (9:1:0.1)) to give heptadecan-9-yl 8- ((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl) amino)propyl)(8-oxo-8-((2- propylnonyl)oxy)octyl)amino)octanoate (405 mg, 57%) as light yellow wax. HPLC/UV (254 nm, Method-A): RT = 7.04 min. MS (CI): m/z (MH + ) 874.7 for C 5 3H99N3O6. 1 H NMR (300 MHz, CDCl 3 ): d 0.86 (m, 12H); 1.22-1.49 (m, 60H); 1.60 (m, 6H); 1.75 (m, 2H); 2.27 (m, 4H); 2.43 (m, 4H); 2.59 (m, 2H); 3.25 (d, 3H, J = 4.9 Hz); 3.63 (m, 2H); 3.96 (d, 2H, J = 5.7 Hz); 4.84 (m, 1H). AF. Compound 10: 2-Butylnonyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methyl amino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)octano ate 5-Methylenedodecane [00418] Same as procedure as for compound 9 but using dodecan-5-one instead of undecan-4-one. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m, 14H); 1.99 (m, 4H); 4.68 (s, 2H). 2-Butylnonan-1-ol [00419] Same as procedure as for compound 9 but using 5-methylenedodecane instead of 4-methyleneundecane. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m, 19H); 3.52 (d, 2H, J = 5.5 Hz). 2-Butylnonyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methyl amino)-3,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate [00420] Same as procedure as for compound 9 but using 2-butylnonan-1-ol instead of 2- propylnonan-1-ol. Light yellow wax. HPLC/UV (254 nm, Method-A): RT = 6.98 min. MS (CI): m/z (MH + ) 888.7 for C 5 4H101N3O6. 1 H NMR (300 MHz, CDCl 3 ): d 0.86 (m, 12H); 1.22-1.49 (m,62H); 1.60 (m, 6H); 1.78 (m, 2H); 2.27 (m, 4H); 2.43 (m, 4H); 2.58 (m, 2H); 3.25 (d, 3H, J = 4.8 Hz); 3.64 (m, 2H); 3.96 (d, 2H, J = 5.8 Hz); 4.84 (m, 1H). AG. Compound 11: 2-Ethylnonyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methyl amino) -3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate 3-Methylenedecane [00421] Same as procedure as for compound 9 but using decan-3-one instead of undecan- 4-one. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m,10H); 1.99 (m, 4H); 4.68 (s, 2H). 2-Ethylnonan-1-ol [00422] Same as procedure as for compound 9 but using 3-methylenedecane instead of 4- methyleneundecane. 1 H NMR (300 MHz, CDCl 3 ): d 0.87 (m, 6H); 1.22-1.36 (m, 15H); 3.52 (d, 2H, J = 5.5 Hz). 2-Ethylnonyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methyl amino) -3,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate [00423] Same as procedure as for compound 9 but using 2-ethylnonan-1-ol instead of 2- propylnonan-1-ol. Light yellow wax. HPLC/UV (254 nm, Method-A): RT = 6.99 min. MS (CI): m/z (MH + ) 860.7 for C 5 2H97N3O6. 1 H NMR (300 MHz, CDCl 3 ): d 0.86 (m, 12H); 1.22-1.49 (m, 58H); 1.60 (m, 5H); 1.78 (m, 2H); 2.26 (m, 4H); 2.43 (m, 4H); 2.58 (m, 2H); 3.25 (d, 3H, J = 4.6 Hz); 3.64 (m, 2H); 3.96 (d, 2H, J = 5.7 Hz); 4.84 (m, 1H); 7.43 (br, 2H). AH. Compound 14: Undecan-3-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(6-((2-octyldecyl)oxy)-6-oxohexyl)amino)octa noate 2-Octyldecanoic acid [00424] A solution of diisopropylamine (2.92 mL, 20.8 mmol) in THF (10 mL) was cooled to -78 °C and a solution of n-BuLi (7.5 mL, 18.9 mmol, 2.5 M in hexanes) was added. The reaction was allowed to warm to 0 °C. To a solution of decanoic acid (2.96 g, 17.2 mmol) and NaH (754 mg, 18.9 mmol, 60% w/w) in THF (20 mL) at 0 °C was added the solution of LDA and the mixture was allowed to stir at room temperature for 30 min. After this time 1- iodooctane (5 g, 20.8 mmol) was added and the reaction mixture was heated at 45 °C for 6 h. The reaction was quenched with 1N HCl (10 mL). The organic layer was dried over MgSO 4 , filtered and evaporated under vacuum. The residue was purified by silica gel chromatography (0-20% ethyl acetate in hexanes) to yield 2-octyldecanoic acid (1.9 g, 6.6 mmol). 1 H NMR (300 MHz, CDCl 3 ) d: ppm 2.38 (br. m, 1H); 1.74-1.03 (br. m, 28H); 0.91 (m, 6H). 2-Octyldecanol [00425] A solution of 2-octyldecanoic acid (746 mg, 2.6 mmol) in dry THF (12 mL) was added to a stirred solution of LAH (5.2 mL, 5.2 mmol, 1M solution in THF) in dry THF (6 mL) under nitrogen at 0 ° C. The reaction was allowed to warm to room temperature and stirred at room temperature for 12 h. A solution of saturated Na2SO4*10H2O solution (10 mL) was added. The solids were filtered through a plug of Celite. The filtrate was evaporated under vacuum and the residue was purified by silica gel chromatography (0-20% ethyl acetate in hexanes) to yield 2-octyldecan-1-ol (635 mg, 2.3 mmol). 1 H NMR (300 MHz, CDCl 3 ) d: ppm 3.55 (d, 2H); 1.57-1.18 (m, 30H); 0.91 (m, 6H). 2-Octyldecyl 6-bromohexanoate [00426] To a solution of 6-bromohexanoic acid (606 mg, 3.1 mmol) and 2-octyldecanol (840 mg, 3.1 mmol) in dichloromethane (3.1 mL) was added N-(3-dimethylaminopropyl)-N¢- ethylcarbodiimide hydrochloride (655 mg, 3.4 mmol), N,N-diisopropylethylamine (1.2 mL, 6.8 mmol) and DMAP (76 mg, 0.62 mmol). The reaction was allowed to stir at room temperature for 16 h. The reaction was diluted with dichloromethane and washed with saturated sodium bicarbonate. The organic layer was separated and washed with brine, and dried over MgSO 4 . The organic layer was filtered and evaporated in vacuo. The residue was purified by silica gel chromatography (0-100% ethyl acetate in hexanes) to obtain 2- octyldecyl 6-bromohexanoate (849 mg, 1.9 mmol, 61%). 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.01 (d, 2H); 3.56 (t, 0.24); 3.43 (t, 1.76); 2.35 (t, 2H); 1.99-1.43 (m, 8H); 1.29 (m, 27H); 0.91 (m, 6H). Undecan-3-yl 8-bromooctanoate [00427] To a solution of 3-undecanol (4.14 g, 24 mmol), 8-bromooctanoic acid (8.01 g, 36 mmol) and DMAP (0.58 g, 4.8 mmol) in dichloromethane (50 mL) at 0 ˚C was added EDCI (6.9 g, 36 mmol) and the reaction mixture stirred at room temperature overnight. TLC showed the reaction completed. The reaction mixture was cooled to 0 ˚C and a solution of hydrochloric acid (10 mL conc. HCl, 90 mL water, 7.5 g sodium chloride) was added very slowly over 20 minutes. Then acetonitrile (100 mL) and hexane (100 mL) were added, the layers separated and the organic layer dried and removed in vacuum to give an oil. The oil was dissolved in hexane (100 mL) and washed with a mixture of acetonitrile (100 mL) and 5% sodium bicarbonate (100 mL). The hexane layer was separated and filtered through Celite, which was then washed with hexane. The solvent was removed under vacuum to give undecan-3-yl 8-bromooctanoate (8.76 g, 97%) as colorless oil. Contains approximately 13% of the corresponding chloride. 1 H NMR (300 MHz, CDCl 3 ): d ppm 4.82-4.76 (m, 1H); 3.39 (t, 2H, J = 6.7 Hz); 2.44 (t, 0.3H, J = 7.4 Hz, for CH 2 Cl); 2.28 (t, 2H, J = 7.5 Hz, for CH 2 Br); 1.88-1.79 (m, 2H); 1.70- 1.42 (m, 6H); 1.38-1.17 (m, 18H); Undecan-3-yl 8-({3-[(tert-butoxycarbonyl)amino]propyl}amino)octanoate [00428] Prepared from tert-butyl N-(3-aminopropyl)carbamate analogously to compound 34 using undecan-3-yl 8-bromooctanoate instead of 4-pentylnonyl-8-bromooctanoate to give undecan-3-yl 8-({3-[(tert-butoxycarbonyl)amino]propyl}amino)octanoate. Undecan-3-yl 8-((3-(((tert-butoxyoxycarbonyl)amino)propyl)(6-((2-octyldec yl)oxy)-6- oxohexyl)amino)octanoate [00429] To a solution of 2-octyldecyl 6-bromohexanoate (0.4 g, 0.894 mmol) and undecan-3-yl 8-({3-[(tert-butoxycarbonyl)amino]propyl}amino)octanoate (0.501 g, 0.894 mmol) in 40 mL of a 1:1 mixture of cyclopropyl methyl ether and acetonitrile were added potassium carbonate (0.741 g, 5.363 mmol) and potassium iodide (0.163 g, 0.983 mmol). The reaction was heated to 77 o C and stirred for 16 hours. The mixture was cooled to room temp., filtered, and the filtrate evaporated under vacuum. The residue was purified by silica gel chromatography (0-100% (solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM) to give undecan-3-yl 8-((3-(((tert-butoxycarbonyl)amino)propyl)(6-((2- octyldecyl)oxy)-6-oxohexyl)amino)octanoate (323 mg, 0.39 mmol, 43%). UPLC/ELSD: RT = 3.15 min. MS (ES): m/z (MH + ) 837.525 for C 5 1H100N2O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.64 (bm, 1H), 4.83 (p, 1H); 3.98 (d, 2H); 3.19 (bm, 2H); 2.55-2.23 (m, 10H); 1.75-1.18 (m, 72H); 0.90 (m, 12H). Undecan-3-yl 8-((3-aminopropyl)(6-((2-octyldecyl)oxy)-6-oxohexyl)amino)oc tanoate [00430] To a solution of undecan-3-yl 8-({3-[(tert-butoxycarbonyl)amino]propyl}({6-[(2- octyldecyl)oxy]-6-oxohexyl})amino)octanoate (0.277 g, 0.331 mmol) in DCM (10 mL) was added trifluoroacetic acid (0.506 mL, 6.62 mmol). The reaction was allowed to stir at RT for 4 h. The reaction was diluted with DCM and slowly quenched with a saturated aqueous sodium bicarbonate solution. The organic layer was separated, washed with brine, dried with Na2SO4, filtered and evaporated under vacuum to give undecan-3-yl 8-((3- aminopropyl)(6-((2-octyldecyl)oxy)-6-oxohexyl)amino)octanoat e (0.24 g, 0.33 mmol, quant.) which was taken to the next step without further purification. UPLC/ELSD: RT = 2.79 min. MS (ES): m/z (MH + ) 737.613 for C 4 6H92N2O4. 3-Methoxy-4-(methylamino)cyclobut-ene-1,2-dione [00431] To a solution of 3,4-dimethoxy-3-cyclobutene-1,2-dione (1 g, 7 mmol) in 100 mL diethyl ether was added a 2M methylamine solution in THF (3.8 mL, 7.6 mmol) and a ppt. formed almost immediately. The mixture was stirred at room temperature for 24 hours, then filtered, the filter solids washed with diethyl ether and air-dried. The filter solids were dissolved in hot EtOAc, filtered, the filtrate allowed to cool to room temp., then cooled to 0 o C to give a ppt. This was isolated via filtration, washed with cold EtOAc, air-dried, then dried under vacuum to give 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (0.70 g, 5 mmol, 73%) as a white solid. 1 H NMR (300 MHz, DMSO-d 6 ) d: ppm 8.50 (br. d, 1H, J = 69 Hz); 4.27 (s, 3H); 3.02 (sdd, 3H, J = 42 Hz, 4.5 Hz). Undecan-3-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)pr opyl)(6-((2- octyldecyl)oxy)-6-oxohexyl)amino)octanoate [00432] 3-Methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (69 mg, 0.49 mmol) was reacted as in Compound 34 using undecan-3-yl 8-((3-aminopropyl)(6-((2-octyldecyl)oxy)-6- oxohexyl)amino)octanoate instead of bis(4-pentylnonyl)-8,8'-((3- aminopropyl)azanediyl)dioctanoate to give undecan-3-yl 8-((3-((2-(methylamino)-3,4- dioxocyclobut-1-en-1-yl)amino)propyl)(6-((2-octyldecyl)oxy)- 6-oxohexyl)amino)octanoate (80 mg, 0.09 mmol, 29%). UPLC/ELSD: RT = 2.99 min. MS (ES): m/z (MH + ) 847.392 for C 51 H 95 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.82 (p, 1H); 3.97 (m, 2H); 3.69 (m, 2H); 3.29 (m, 3H); -2.62-2.25 (m, 10H); 1.85-1.17 (m, 64H); 0.91 (m, 12H). AI. Compound 15: Undecan-3-yl 8-((6-((2-hexyldecyl)oxy)-6-oxohexyl)(3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino )octanoate [00433] Compound 15 was prepared analogously to compound 14 starting from decanoic acid and using 1-iodohexane instead of 1-iodooctane. UPLC/ELSD: RT = 2.86 min. MS (ES): m/z (MH + ) 819.269 for C 4 9H91N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.78 (m, 1H); 3.95 (m, 2H); 3.63 (m, 2H); 3.28 (m, 3H); 2.59-2.20 (m, 13H); 1.83-1.16 (m, 58H); 0.87 (m, 12H). AJ. Compound 16: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((2-pentylnonyl)oxy)octyl)amino)oct anoate
[00434] To a solution of 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2-(methylamino)- 3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}octanoic acid (400 mg, 0.56 mmol) and 2- pentylnonan-1-ol (364 mg, 1.7 mmol) in 5.6 mL dry DCM was added 4- (dimethylamino)pyridine (35.0 mg, 0.28 mmol) and dicyclohexyl carbodiimide (369 mg, 1.7 mmol). The solution was stirred at room temperature for 18 hours, after which no starting alcohol remained by LC/MS. The mixture was diluted with a DCM and washed twice with aqueous saturated sodium bicarbonate solution, dried (MgSO 4 ), filtered and conc. The residue was purified by silica gel chromatography (0-50% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-((3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)(8-ox o-8-((2- pentylnonyl)oxy)octyl)amino)octanoate (154 mg, 0.17 mmol, 30%) as a white waxy solid. UPLC/ELSD: RT = 3.04 min. MS (ES): m/z (MH + ) 903.8 for C 55 H 103 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.32 (s, 1H); 4.87 (quint., 1H, J = 6 Hz); 4.00 (d, 2H, J = 6 Hz); 3.68 (br. s, 2H); 3.29 (d, 3H, J = 6 Hz); 2.65 (br. s, 2H); 2.51 (br. m, 4H); 2.33 (m, 5H); 1.81 (br. m, 3H); 1.64 (br. m, 6H); 1.52 (br. m, 9H); 1.28 (br. s, 61H); 0.88 (br. t, 13H, J = 7.5 Hz). AK. Compound 17: Heptadecan-9-yl 8-({8-[(3-isopropylnonyl)oxy]-8-oxooctyl}(3-{[2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino )octanoate [00435] To a solution of 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2-(methylamino)- 3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}octanoic acid (400 mg, 0.57 mmol) and 3- isopropylnonan-1-ol (317 mg, 1.7 mmol) in 5.6 mL dry DCM was added 4- (dimethylamino)pyridine (35.0 mg, 0.28 mmol) and dicyclohexylcarbodiimide (369 mg, 1.7 mmol). The solution was stirred at room temperature for 18 hours, after which no starting alcohol remained by LC/MS. The mixture was diluted with a DCM and washed twice with aqueous saturated sodium bicarbonate solution, dried (MgSO 4 ), filtered and conc. The residue was purified by silica gel chromatography (0-50% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-({8-[(3- isopropylnonyl)oxy]-8-oxooctyl}(3-{[2-(methylamino)-3,4-diox ocyclobut-1-en-1- yl]amino}propyl)amino)octanoate (202 mg, 0.23 mmol, 41%) as a white waxy solid. UPLC/ELSD: RT = 2.92 min. MS (ES): m/z (MH + ) 875.5 for C 53 H 99 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.32 (s, 1H); 4.87 (quint., 1H, J = 6 Hz); 4.08 (m, 2H, J = 7 Hz); 3.67 (br. s, 2H); 3.30 (d, 3H, J = 5 Hz); 2.64 (br. s, 2H); 2.51 (br. m, 4H); 2.31 (m, 5H); 1.81 (br. m, 3H); 1.64 (br. m, 6H); 1.52 (br. m, 9H); 1.28 (br. s, 46H); 0.90 (br. m, 15H). AL. Compound 18: Heptadecan-9-yl 8-[(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl]amino}propyl)({8-oxo-8-[(3-propylnonyl)oxy]octyl})amino]o ctanoate [00436] To a solution of 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2-(methylamino)- 3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}octanoic acid (400 mg, 0.57 mmol) and 3- propylnonan-1-ol (317 mg, 1.7 mmol) in 5.6 mL dry DCM was added 4- (dimethylamino)pyridine (35.0 mg, 0.28 mmol) and dicyclohexylcarbodiimide (369 mg, 1.7 mmol). The solution was stirred at room temperature for 18 hours, after which no starting alcohol remained by LC/MS. The mixture was diluted with a DCM and washed twice with aqueous saturated sodium bicarbonate solution, dried (MgSO 4 ), filtered and conc. The residue was purified by silica gel chromatography (0-50% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-[(3-{[2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)({8-o xo-8-[(3- propylnonyl)oxy]octyl})amino]octanoate (206 mg, 0.24 mmol, 42%) as a white waxy solid. UPLC/ELSD: RT = 2.93 min. MS (ES): m/z (MH + ) 875.5 for C 53 H 99 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.32 (s, 2H); 4.87 (quint., 1H, J = 5 Hz); 4.10 (t, 2H, J = 8 Hz); 3.68 (br. s, 2H); 3.30 (d, 3H, J = 5 Hz); 2.67 (br. s, 2H); 2.53 (br. m, 4H); 2.31 (t, 5H, J = 7 Hz); 1.83 (br. m, 3H); 1.62 (br. m, 7H); 1.51 (br. m, 10H); 1.28 (br. s, 52H); 0.90 (br. m, 13H). AM. Compound 19: Heptadecan-9-yl 8-({8-[(2-isopropylnonyl)oxy]-8-oxooctyl}(3-{[2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino )octanoate [00437] To a solution of 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2-(methylamino)- 3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}octanoic acid (400 mg, 0.57 mmol) and 3- propylnonan-1-ol (317 mg, 1.7 mmol) in 5.6 mL dry DCM was added 4- (dimethylamino)pyridine (35.0 mg, 0.28 mmol) and dicyclohexylcarbodiimide (369 mg, 1.7 mmol). The solution was stirred at room temperature for 18 hours, after which no starting alcohol remained by LC/MS. The mixture was diluted with a DCM and washed twice with aqueous saturated sodium bicarbonate solution, dried (MgSO 4 ), filtered and conc. The residue was purified by silica gel chromatography (0-50% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-({8-[(2- isopropylnonyl)oxy]-8-oxooctyl}(3-{[2-(methylamino)-3,4-diox ocyclobut-1-en-1- yl]amino}propyl)amino)octanoate (163 mg, 0.19 mmol, 33%) as a white waxy solid. UPLC/ELSD: RT = 2.95 min. MS (ES): m/z (MH + ) 875.5 for C 5 3H99N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.23 (s, 1H); 4.87 (quint., 1H, J = 6 Hz); 3.94 (m, 2H, J = 5 Hz); 3.60 (br. s, 2H); 3.20 (d, 3H, J = 6 Hz); 2.94 (s, 1H); 2.57 (br. s, 2H); 2.43 (br. m, 4H); 2.23 (br. m, 5H); 1.73 (br. m, 4H); 1.55 (br. m, 4H); 1.41 (br. m, 9H); 1.19 (br. s, 45H); 0.81 (br. m, 14H). AN. Compound 20: 3-Butylnonyl 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino }octanoate [00438] To a solution of 8-{[8-(heptadecan-9-yloxy)-8-oxooctyl](3-{[2-(methylamino)- 3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}octanoic acid (400 mg, 0.57 mmol) and 3- butylnonan-1-ol (341 mg, 1.7 mmol) in 5.6 mL dry DCM was added 4- (dimethylamino)pyridine (35.0 mg, 0.28 mmol) and dicyclohexylcarbodiimide (369 mg, 1.7 mmol). The solution was stirred at room temperature for 18 hours, after which no starting alcohol remained by LC/MS. The mixture was diluted with a DCM and washed twice with aqueous saturated sodium bicarbonate solution, dried (MgSO 4 ), filtered and conc. The residue was purified by silica gel chromatography (0-50% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylnonyl 8-{[8-(heptadecan-9- yloxy)-8-oxooctyl](3-{[2-(methylamino)-3,4-dioxocyclobut-1-e n-1- yl]amino}propyl)amino}octanoate (116 mg, 0.13 mmol, 23%) as a white waxy solid. UPLC/ELSD: RT = 3.01 min. MS (ES): m/z (MH + ) 888.7 for C 5 4H101N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.23 (s, 1H); 4.78 (quint., 1H, J = 6 Hz); 4.01 (m, 2H, J = 5 Hz); 3.58 (br. s, 2H); 3.20 (d, 3H, J = 6 Hz); 2.50 (br. s, 2H); 2.37 (br. m, 4H); 2.22 (br. t, 4H); 1.69 (br. m, 5H); 1.55 (br. m, 6H); 1.43 (br. m, 5H); 1.35 (br. m, 5H); 1.19 (br. s, 50H); 0.81 (br. m, 12H). AO. Compound 21: Heptadecan-9-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(2- hydroxyethyl)amino) octanoate Molecular Weight: 794.34 [00439] UPLC/ELSD: RT = 3.20 min. MS (ES): m/z (MH + ) 795.093 for C 50 H 99 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.10 (t, 2H); 3.56 (m, 2H); 2.67-2.41 (m, 6H); 2.30 (m, 4H), 1.74-1.17 (m, 72H); 0.90 (m, 12H). AP. Compound 22: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(2- hydroxyethyl)amino)octanoate Step 1: Heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate [00440] A solution of heptadecan-9-yl 8-bromooctanoate (10 g, 21.67 mmol) and ethanolamine (39.70 g, 649.96 mmol) in EtOH (5 mL) was heated to 65 °C for 16h. The reaction was cooled to rt and dissolved in ethyl acetate and extracted with water (4X). The organic layer was separated, washed with brine, dried with Na 2 SO 4 , filtered and evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0- 100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM to obtain heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate (7.85 g, 82%). UPLC/ELSD: RT = 2.06 min. MS (ES): m/z (MH + ) 442.689 for C 27 H 55 NO 3. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 3.66 (t, 2H); 2.79 (t, 2H); 2.63 (m, 2H); 2.30 (t, 2H); 1.77- 1.20 (m, 40H); 0.90 (m, 6H). Step 2: Compound 22: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(2- hydroxyethyl)amino)octanoate [00441] To a solution of 3-butylheptyl 8-bromooctanoate (6.15 g, 16.31 mmol) and heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate (6.86 g, 15.53 mmol) in a mixture of CPME (15 mL) and acetonitrile (6 mL) was added potassium carbonate (8.59 g, 62.12 mmol) and potassium iodide (2.84 g, 17.08 mmol). The reaction was allowed to stir at 77 °C for 16 h. The reaction was cooled and filtered, and the volatiles were evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0- 100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM to obtain 3-butylheptyl 8- ((8-(heptadecan-9-yloxy)-8-oxooctyl)(2-hydroxyethyl)amino)oc tanoate (4.53 g, 37.8%). Chemical Formula: C 46 H 91 NO 5 Molecular Weight: 738.24 [00442] UPLC/ELSD: RT = 3.04 min. MS (ES): m/z (MH + ) 739.464 for C 46 H 91 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.11 (m, 2H), 3.57 (bm, 2H); 2.73-2.39 (m, 6H); 2.30 (m, 4H); 1.72-1.17 (m, 64H); 0.92 (m, 12H). AQ. Compound 23: Bis(3-hexylnonyl) 8,8'-((2-hydroxyethyl)azanediyl)dioctanoate Chemical Formula: C 48 H 95 NO 5 Molecular Weight: 766.29 [00443] UPLC/ELSD: RT = 3.11 min. MS (ES): m/z (MH + ) 767.710 for C 48 H 95 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.10 (m, 4H); 3.55 (bm, 2H); 2.65-2.37 (m, 6H); 2.31 (m, 4H); 1.72-1.16 (m, 67H); 0.91 (m, 12H). AR. Compound 24: Heptadecan-9-yl 8-((2-hydroxyethyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino) octanoate Chemical Formula: C 48 H 95 NO 5 Molecular Weight: 766.29 [00444] UPLC/ELSD: RT = 3.13 min. MS (ES): m/z (MH + ) 767.586 for C 4 8H95NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.10 (t, 2H); 3.56 (bm, 2H), 2.68-2.40 (m, 6H); 2.30 (m, 4H); 1.74-1.18 (m, 68H); 0.91 (m, 12H). AS. Compound 25: Bis(3-pentyloctyl) 8,8'-((2-hydroxyethyl)azanediyl)dioctanoate Chemical Formula: C 4 4H87NO5 Molecular Weight: 710.18 [00445] UPLC/ELSD: RT = 2.86 min. MS (ES): m/z (MH + ) 711.341 for C 4 4H87NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.10 (t, 4H); 3.55 (m, 2H); 2.65-2.38 (m, 6H), 2.31 (m, 4H); 1.73-1.16 (m, 59); 0.91 (m, 12H). AT. Compound 26: Heptadecan-9-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino )octanoate Ethyl 3-hexylnon-2-enoate [00446] Triethyl phosphonoacetate (26.33 g, 117.4 mmol) was added dropwise over 20 minutes to a suspension of sodium hydride (4.697 g, 117.4 mmol) in THF (294 mL) and the mixture was stirred at room temperature until gas evolution ceased (approximately 30 min). The reaction mixture was chilled to 0 °C and 7-tridecanone (10 g, 58.7 mmol) was added. The reaction was gradually warmed to room temperature, then heated to reflux and stirred overnight. The reaction was quenched with saturated aqueous sodium bicarbonate. The aqueous phase was extracted with diethyl ether, and the organic extracts were washed with brine, dried with MgSO 4 , and concentrated. The crude material was purified by silica gel chromatography (0-20% EtOAc:hexanes) to afford ethyl 3-hexylnon-2-enoate (6.7 g, 27.9 mmol, 47.5%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.63 (s, 1H); 4.15 (q, 2H); 2.61 (t, 2H); 2.15 (t, 2H); 1.53-1.20 (m, 19H); 0.91 (m, 6H). Ethyl 3-hexylnonanoate [00447] To a flask containing a slurry of Pearlmans catalyst (0.73 g, 5.2 mmol) in ethanol (20 mL) under N2 was added a solution of ethyl 3-hexylnon-2-enoate (6.975 g, 25.9 mmol) in ethanol (5 mL). The reaction was stirred under H 2 (balloon) for 16h. The reaction was filtered through a plug of Celite and the filtrate was evaporated under vacuum to afford ethyl 3-hexylnonanoate (6.7 g, 24.7 mmol, 95 %). The residue was taken to the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.16 (q, 2H); 2.23 (d, 2H); 1.86 (bs, 1H); 1.28 (m, 23H); 0.90 (m, 6H). 3-Hexylnonan-1-ol [00448] To a solution of lithium aluminum hydride (49.5 mL of 1M solution in THF, 49.5 mmol) in THF was added a solution of ethyl 3-hexylnonanoate (6.7 g, 24.7 mmol) in THF (20 mL). The reaction was stirred at room temperature for 16h. The reaction was quenched with a saturated solution of sodium sulfate decahydrate. The white solids were removed by filtration through a plug of Celite and the filtrate was evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0- 100% ethyl acetate in hexanes to obtain 3-hexylnonan-1-ol (5.62 g, 24.6 mmol). Heptadecan-9-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(3-((2-(methylamino)-3, 4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate [00449] Heptadecan-9-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(3-((2-(methylamino)-3, 4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoate was prepared analogously to compound 9 using 3-hexylnonan-1-ol instead of 2-propylnonan-1-ol. UPLC/ELSD: RT = 3.17 min. MS (ES): m/z (MH + ) 917.085 for C 58 H 105 N 3 O 6 . . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.68 (bm, 2H); 3.28 (m; 3H); 2.75-2.44 (m, 6H); 2.31 (m, 4H); 1.82 (m, 3H); 1.72-1.18 (m, 71H); 0.90 (m, 12H). AU. Compound 27: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino )octanoate Step 1: Heptadecan-9-yl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate [00450] A solution of tert-butyl N-(3-aminopropyl)carbamate (34.35 g, 197.15 mmol) in EtOH (200 mL) was heated to 65 °C and a solution of heptadecan-9-yl 8- bromooctanoate (26 g, 56.33 mmol) in EtOH (90 mL) was added over 3 h. The reaction was heated at 65 °C for 3h. The reaction was cooled to <50 °C and EtOH was evaporated under vacuum and azeotroped with heptane (4X). To a solution of crude product in 2-MeTHF (150 mL) 5% K 2 CO 3 (150 mL) was added and the resulting mixture was stirred for 10 minutes. The two layers were allowed to form. The aqueous layer was removed and the 2-MeTHF layer was washed with 100 mL water (x3). The organic layer was separated, washed with brine, dried with Na 2 SO 4 , filtered and evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0-100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM to obtain heptadecan-9-yl 8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate (20g, 63.9%). UPLC/ELSD: RT = 2.34 min. MS (ES): m/z (MH + ) 555.319 for C 33 H 66 N 2 O 4. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.18 (bs, 1H); 4.89 (p, 1H); 3.22 (m, 2H); 2.64 (t, 2H); 2.59 (t, 2H); 2.30 (t, 2H); 1.73-1.21 (m, 50H); 0.90 (m, 6H). Step 2: 3-Butylheptyl 8-((3-((tert-butoxycarbonyl)amino)propyl)(8-(heptadecan-9-yl oxy)-8- oxooctyl)amino)octanoate [00451] To a solution of heptadecan-9-yl 8-({3-[(tert- butoxycarbonyl)amino]propyl}amino)octanoate (11.76 g, 21.19 mmol) and 3-butylheptyl 8- bromooctanoate (9.2 g, 24.37 mmol) in propionitrile (52 mL) was added Potassium carbonate (4.39 g, 31.79 mmol) and Potassium iodide (0.53 g, 3.18 mmol). The reaction was heated at 80 °C for 16h. The reaction was cooled and filtered, and the volatiles were evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0-100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM to obtain 3-butylheptyl 8-((3-((tert- butoxycarbonyl)amino)propyl)(8-(heptadecan-9-yloxy)-8-oxooct yl)amino)octanoate (9.68 g, 53.6%). UPLC/ELSD: RT = 3.07 min. MS (ES): m/z (MH + ) 851.216 for C 5 2H102N2O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.68 (bs, 1H); 4.90 (p, 1H); 4.11 (t, 2H); 3.20 (m, 2H); 2.52-2.24 (m, 10H); 1.76-1.20 (m, 74H); 0.90 (m, 12H). Step 3: 3-Butylheptyl 8-((3-aminopropyl)(8-(heptadecan-9-yloxy)-8- oxooctyl)amino)octanoate [00452] To a solution of 3-butylheptyl 8-({3-[(tert-butoxycarbonyl)amino]propyl}[8- (heptadecan-9-yloxy)-8-oxooctyl]amino)octanoate (7 g, 8.22 mmol) in DCM (25 mL) was added trifluoroacetic acid (9.4 mL, 123.32 mmol). The reaction was allowed to stir at rt for 2 h. The reaction was evaporated under vacuum. The residue was dissolved in mixture of methyl THF/heptane (1:9) and extracted with sat. sodium bicarbonate (3X). The organic layer was separated, washed with brine, dried with Na 2 SO 4 , filtered and evaporated under vacuum to obtain 3-Butylheptyl 8-((3-aminopropyl)(8-(heptadecan-9-yloxy)-8- oxooctyl)amino)octanoate. This was taken as a crude to the next step without further purification. UPLC/ELSD: RT = 2.63 min. MS (ES): m/z (MH + ) 751.305 for C 4 7H94N2O4. Step 4: Compound 27: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino )octanoate [00453] To a solution of 3-butylheptyl 8-[(3-aminopropyl)[8-(heptadecan-9-yloxy)-8- oxooctyl]amino]octanoate (7 g, 9.32 mmol) in methyl THF (31 mL) was added 3-methoxy-4- (methylamino)cyclobut-3-ene-1,2-dione (1.71 g, 12.11 mmol), and a aqueous solution of 10% Sodium bicarbonate (8.6 mL, 10.25 mmol). The reaction was allowed to stir at 50 °C for 2.5 h. The reaction was cooled to rt and diluted with heptane and extracted with water. The organic layer was separated, washed with brine, dried with Na2SO4, filtered and evaporated under vacuum. The residue was purified by flash chromatography (ISCO) by 0- 100% (a solution of 20% MeOH, 80% DCM, 1% NH 4 OH) in DCM to obtain 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4-dioxocyclobut-1-en-1- yl)amino)propyl)amino)octanoate (5.4 g, 63%). UPLC/ELSD: RT = 2.98 min. MS (ES): m/z (MH + ) 861.714 for C 5 2H97N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.10 (t, 2H); 3.75 (m, 2H); 3.39-3.20 (m, 5H); 3.08 (m, 4H); 2.31 (m, 4H); 2.12 (bm, 2H); 1.81-1.20 (m, 65H); 0.90 (m, 12H). AV. Compound 28: Bis(3-hexylnonyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate [00454] Compound 28 was prepared analogously to compound 34 starting from 3- hexylnonyl-8-bromooctanoate instead of 4-pentylnonyl-8-bromooctanoate. UPLC/ELSD: RT = 3.08 min. MS (ES): m/z (MH + ) 889.332 for C 5 4H101N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.10 (m, 4H); 3.67 (bm, 2H); 3.28 (m, 3H); 2.57 (m, 2H); 2.44 (m, 4H); 2.31 (m, 4H); 1.77 (m, 2H); 1.62 (m, 8H); 1.52-1.19 (m, 58H); 0.90 (m, 12H). AW. Compound 29: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en- 1-yl)amino)propyl)(8-oxo-8-((3-pentyloctyl)oxy)octyl)amino)o ctanoate [00455] Compound 29 was prepared analogously to compound 9 starting from 6- undecanone instead of 7-tridecanone. UPLC/ELSD: RT = 3.12 min. MS (ES): m/z (MH + ) 889.332 for C 54 H 101 N 3 O 6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (p, 1H); 4.10 (t, 2H); 3.67 (bm, 2H); 3.29 (m, 3H); 2.67-2.41 (m, 6H); 2.31 (m, 4H); 1.85-1.18 (m, 71H), 0.90 (m, 12H). AX. Compound 30: Bis(3-pentyloctyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate Step 1: Bis(3-pentyloctyl) 8,8'-((3-((tert-butoxycarbonyl)amino)propyl) azanediyl)dioctanoate [00456] To a solution of 3-pentyloctyl 8-bromooctanoate (5.61 g, 13.8 mmol) and 3- pentyloctyl 8-((3-((tert-butoxycarbonyl)amino)propyl)amino)octanoate (6.00 g, 12.0 mmol) in propionitrile (30 mL) was added potassium carbonate (2.49 g, 18.0 mmol) and iodopotassium (300 mg, 1.80 mmol). The reaction was allowed to stir at 80 ºC for 16 h. Upon cooling to room temperature, the reaction mixture was filtered via vacuum filtration. The residue in the vessel and the filter cake on the funnel was washed twice with propionitrile. The filtrate was then concentrated in vacuo at 40 °C. The crude residue was purified by silica gel chromatography (0-5-10-20-25-30-35-40-50-80-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3- pentyloctyl) 8,8'-((3-((tert-butoxycarbonyl)amino)propyl) azanediyl)dioctanoate (7.37 g, 8.95 mmol, 74%) as a light yellow transparent oil. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.66 (br. s, 1H); 4.08 (t, 4H, J = 6.0 Hz); 3.17 (br. q, 2H, J = 6.0 Hz); 2.43 (t, 2H, J = 6.0 Hz); 2.34 (br. t, 4H, J = 6.0 Hz); 2.28 (t, 4H, J = 9.0 Hz); 1.67-1.52 (m, 10H); 1.48-1.37 (m, 14H); 1.35-1.17 (m, 45H); 0.88 (t, 12H, J = 6.0 Hz). Step 2: Bis(3-pentyloctyl) 8,8'-((3-aminopropyl)azanediyl)dioctanoate [00457] To a round bottom flask equipped with a stir bar was added bis(3-pentyloctyl) 8,8'-((3-((tert-butoxycarbonyl)amino)propyl) azanediyl)dioctanoate (3.00 g, 3.64 mmol). The oil was dissolved in cyclopentyl methyl ether (8 mL) and stirred for 5 minutes.3M HCl in cyclopentyl methyl ether (6.07 mL, 18.2 mmol) was added dropwise. After addition was complete, the reaction was heated to 40 ºC for 1 hour and reaction completion was monitored by TLC/LCMS analysis. The reaction was cooled to room temperature, and then chilled to 0 °C.10% K 2 CO 3 solution was then added dropwise to the reaction mixture. After addition was complete, the aqueous/cyclopentyl methyl ether emulsion was diluted with EtOAc and the resulting mixture stirred for 10 minutes. The solution was transferred to a separation funnel and the layers were separated. The organic layer was dried (MgSO 4 ), filtered, and concentrated. The residue was redissolved in heptane and washed twice with MeCN. The heptane layer was dried (MgSO 4 ), filtered, and concentrated to afford crude bis(3- pentyloctyl) 8,8'-((3-aminopropyl)azanediyl)dioctanoate (2.43 g, 3.36 mmol, 92%) as an off- white oil. The crude material was carried onto the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 4H, J = 6.0 Hz); 2.98 (t, 2H, J = 6.0 Hz); 2.71 (t, 2H, J = 6.0 Hz); 2.54 (br. t, 4H, J = 6.0 Hz); 2.28 (t, 6H, J = 6.0 Hz); 1.76 (br. pentet, 2H, J = 2.0 Hz); 1.66-1.52 (m, 9H); 1.52-1.43 (m, 4H); 1.37-1.18 (m, 45H); 0.88 (t, 12H, J = 6.0 Hz). Step 3: Compound 30: Bis(3-pentyloctyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate [00458] To a round bottom flask equipped with a stir bar was added bis(3-pentyloctyl) 8,8'-((3-aminopropyl)azanediyl)dioctanoate (2.43 g, 3.36 mmol), 3-methoxy-4- (methylamino)cyclobut-3-ene-1,2-dione (616 mg, 4.36 mmol) and 2-Methyl THF (10 mL). 10% K 2 CO 3 solution (10 mL) was added and the resulting biphasic mixture was heated to 45 °C and stirred vigorously for 3 hours. Reaction completion was monitored by TLC/LCMS analysis. Upon completion the mixture was allowed to cool to room temperature. The reaction was diluted with water, layers were separated, and the aqueous layer was extracted twice with heptane. The organics were combined, washed with water (3x), brine, and with a 1:1 acetonitrile/water mixture. The combined organics were then dried (Na2SO4), filtered, and concentrated. The crude residue was azeotroped and concentrated with DCM and MeOH three times to yield a pale yellow crude waxy oil. The crude residue was purified by silica gel chromatography (0-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-pentyloctyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate (2.11 g, 2.54 mmol, 76%) as a white waxy solid. UPLC/ELSD: RT = 2.79 min. MS (ES): m/z (MH + ) 832.34 for C 5 0H93N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.83 (br. s, 1H); 7.61 (br. s, 1H); 4.03 (t, 4H, J = 9.0 Hz); 3.64 (br. s, 2H); 3.28 (br. d, 3H, J = 6.0 Hz); 2.46 (t, 2H, J = 9.0 Hz); 2.33 (br. t, 4H, J = 6.0 Hz); 2.33 (t, 4H, J = 9.0 Hz); 1.74 (br. pentet, 2H, J = 6.0 Hz); 1.62-1.47 (m, 8H); 1.41-1.12 (m, 50H); 0.83 (t, 12H, J = 9.0 Hz). AY. Compound 31: Heptadecan-9-yl 8-((2-hydroxyethyl)(8-oxo-8-((4- pentylnonyl)oxy)octyl)amino)octanoate [00459] To a solution of 4-pentylnonyl-8-bromooctanoate (499 mg, 1.19 mmol) and heptadecan-9-yl 8-((2-hydroxyethyl)amino)octanoate (500 mg, 1.13 mmol) in cyclopentyl methyl ether (5 mL) and acetonitrile (5 mL) was added potassium carbonate (939 mg, 6.79 mmol) and potassium iodide (207 mg, 1.25 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-((2-hydroxyethyl)(8-oxo-8- ((4-pentylnonyl)oxy)octyl)amino)octanoate (242 mg, 0.31 mmol, 27%) as a clear viscous oil. UPLC/ELSD: RT = 3.18 min. MS (ES): m/z (MH + ) 780.66 for C 4 9H97NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.83 (pent., 1H, J = 6.0 Hz); 4.00 (t, 2H, J = 6.0 Hz); 3.64 (br. s, 1H), 3.53 (t, 2H, J = 6.0 Hz); 2.59 (t, 2H, J = 3.0 Hz); 2.46 (br. t, 4H, J = 6.0 Hz); 2.30-2.18 (m, 4H); 1.65-1.37 (m, 15H); 1.36-1.11 (m, 55H); 0.91-0.78 (m, 12H). AZ. Compound 32: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((4-pentylnonyl)oxy)octyl)amino)oct anoate [00460] To a solution of 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3-((2-(methylamino)-3 ,4- dioxocyclobut-1-en-1-yl)amino)propyl)amino)octanoic acid (706 mg, 1.00 mmol), 4- pentylnonan-1-ol (322 mg, 1.50 mmol), and DMAP (37.0 mg, 0.30 mmol) in dichloromethane (5 mL) at room temperature was added N,N’-dicyclohexylcarbodiimide (309 mg, 1.50 mmol). The resulting mixture was allowed to stir at room temperature for 16 h. The reaction was then diluted with additional dichloromethane and washed with saturated aqueous sodium bicarbonate. The organic layer washed with 1M HCl, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5- 10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en- 1-yl)amino)propyl)(8-oxo-8-((4-pentylnonyl)oxy)octyl)amino)o ctanoate (186 mg, 0.21 mmol, 21%) as a beige oil. UPLC/ELSD: RT = 3.15 min. MS (ES): m/z (MH + ) 902.65 for C 5 5H103N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 8.78 (br. s, 1H); 8.18 (br. s, 1H); 4.83 (pent., 2H, J = 6 Hz); 4.01 (t, 2H, J = 6 Hz); 3.68 (br. d, 2H, J = 6 Hz); 3.28 (d, 3H, J = 6 Hz); 3.15 (br. s, 2H); 2.95 (br. s, 4H); 2.32-2.20 (m, 4H); 2.12-1.97 (br. m, 2H); 1.73-1.41 (m, 14H); 1.40-1.11 (m, 54H); 0.93-0.78 (m, 12H). BA. Compound 33: Bis(4-pentylnonyl)-8,8'-((2-hydroxyethyl)azanediyl)dioctanoa te Bis(4-pentylnonyl)-8,8'-((2-hydroxyethyl)azanediyl)dioctanoa te (Compound 33) [00461] To a solution of 4-pentylnonyl-8-bromooctanoate (337 mg, 0.80 mmol) and 4- pentylnonyl-8-((2-hydroxyethyl)amino)octanoate (306 mg, 0.77 mmol) in cyclopentyl methyl ether (4 mL) and acetonitrile (4 mL) was added potassium carbonate (635 mg, 4.59 mmol) and potassium iodide (140 mg, 0.84 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(4-pentylnonyl)-8,8'-((2- hydroxyethyl)azanediyl)dioctanoate (224 mg, 0.30 mmol, 40%) as a clear oil. UPLC/ELSD: RT = 3.05 min. MS (ES): m/z (MH + ) 738.48 for C 46 H 91 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.02 (t, 4H, J = 6.0 Hz); 3.51 (t, 2H, J = 6.0 Hz); 3.25 (br. s, 1H); 2.56 (t, 2H, J = 6.0 Hz); 2.43 (br. t, 4H, J = 6.0 Hz); 2.27 (t, 4H, J = 6.0 Hz); 1.67-1.50 (m, 8H); 1.48-1.36 (m, 5H); 1.36-1.12 (m, 50H); 0.93-0.80 (m, 12H). BB. Compound 34: Bis(4-pentylnonyl)-8,8'-((3-((2-(methylamino)-3,4-dioxocyclo but-1- en-1-yl)amino)propyl)azanediyl)dioctanoate 4-Pentylnonyl-8-((3-((tert-butoxycarbonyl)amino)propyl)amino ) octanoate [00462] To a solution of tert-butyl N-(3-aminopropyl)carbamate (2.99 g, 17.2 mmol) in ethanol (8 mL) was added a solution of 4-pentylnonyl-8-bromooctanoate (1.2 g, 2.86 mmol) in ethanol (7 mL) at room temperature over the course of 20 min. The reaction was heated to 60 °C, and allowed to stir at this temperature for 16 h. The reaction mixture was then cooled to room temperature, and the solvents were evaporated. The resulting residue was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate and brine (5x) until no white precipitate was observed in the aqueous layer. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 4-pentylnonyl-8-((3-((tert- butoxycarbonyl)amino)propyl)amino)octanoate (802 mg, 1.56 mmol, 55%) as a clear oil. UPLC/ELSD: RT = 2.11 min. MS (ES): m/z (MH + ) 513.26 for C 3 0H60N2O4. Bis(4-pentylnonyl)-8,8'-((3-((tert-butoxycarbonyl)amino)prop yl) azanediyl)dioctanoate [00463] To a solution of 4-pentylnonyl-8-bromooctanoate (667 mg, 1.59 mmol) and 4- pentylnonyl-8-((3-((tert-butoxycarbonyl)amino)propyl)amino)o ctanoate (777 mg, 1.52 mmol) in cyclopentyl methyl ether (7 mL) and acetonitrile (7 mL) was added potassium carbonate (1.26 g, 9.09 mmol) and potassium iodide (277 mg, 1.67 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(4-pentylnonyl)-8,8'-((3-((tert- butoxycarbonyl)amino)propyl) azanediyl)dioctanoate (742 mg, 0.87 mmol, 57%) as a clear oil. UPLC/ELSD: RT = 3.11 min. MS (ES): m/z (MH + ) 851.59 for C 52 H 102 N 2 O 6 . Bis(4-pentylnonyl)-8,8'-((3-aminopropyl)azanediyl)dioctanoat e [00464] To a solution of bis(4-pentylnonyl)-8,8'-((3-((tert-butoxycarbonyl)amino)prop yl) azanediyl)dioctanoate (742 mg, 0.87 mmol) in dichloromethane (17 mL) was added trifluoroacetic acid (1.33 mL, 17.4 mmol). The resulting mixture was allowed to stir at room temperature for 4 h prior to being quenched with saturated sodium bicarbonate. The aqueous layer was extracted with dichloromethane. The combined organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(4-pentylnonyl)-8,8'-((3- aminopropyl)azanediyl)dioctanoate (414 mg, 0.55 mmol, 63%) as a clear oil. UPLC/ELSD: RT = 2.58 min. MS (ES): m/z (MH + ) 751.68 for C 47 H 94 N 2 O 4 . Bis(4-pentylnonyl)-8,8'-((3-((2-(methylamino)-3,4-dioxocyclo but-1-en-1- yl)amino)propyl)azanediyl)dioctanoate [00465] To a solution of bis(4-pentylnonyl)-8,8'-((3-aminopropyl)azanediyl)dioctanoat e (414 mg, 0.55 mmol) in tetrahydrofuran (4 mL) and water (750 mL) was added 3-methoxy-4- (methylamino)cyclobut-3-ene-1,2-dione (117 mg, 0.83 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(4-pentylnonyl)-8,8'-((3-((2-(methylamino)-3,4-dioxocyclo but-1-en-1- yl)amino)propyl)azanediyl)dioctanoate (179 mg, 0.21 mmol, 38%) as an off-white waxy solid. UPLC/ELSD: RT = 2.97 min. MS (ES): m/z (MH + ) 860.59 for C 5 2H97N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.29 (br. s, 1H); 4.03 (t, 4H, J = 6 Hz); 3.64 (br. s, 2H); 3.27 (d, 3H, J = 3 Hz); 2.54 (br. t, 2H, J = 6 Hz); 2.41 (br. t, 4H, J = 6 Hz); 2.29 (t, 4H, J = 9 Hz); 1.81-1.69 (m, 2H); 1.68-1.51 (m, 8H); 1.48-1.13 (m, 55H); 0.94-0.80 (m, 12H). BC. Compound 35: 2-(4-Ethylcyclohexyl)ethyl 8-((8-(heptadecan-9-yloxy)-8- oxooctyl)(2-hydroxyethyl)amino)octanoate Chemical Formula: C 4 5H87NO5 Molecular Weight: 722.19 [00466] UPLC/ELSD: RT = 2.92 min. MS (ES): m/z (MH + ) 723.059 for C 4 5H87NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89(p, 1H); 4.12 (m, 2H); 3.57 (bm, 2H); 2.73-2.22 (m, 10H); 1.82-1.15 (m, 61H), 0.90 (m, 11H). BD. Compound 37: 2-(3-Ethylcyclopentyl)ethyl 8-((8-(heptadecan-9-yloxy)-8- oxooctyl)(2-hydroxyethyl)amino)octanoate Chemical Formula: C 4 4H85NO5 Molecular Weight: 708.17 [00467] UPLC/ELSD: RT = 2.89 min. MS (ES): m/z (MH + ) 709.614 for C 4 4H85NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.09 (m, 2H); 3.56 (m, 2H); 2.66-2.40 (m, 6H); 2.30 (m, 4H); 2.06-1.05 (m, 60H), 0.90 (m, 10H). BE. Compound 39: Heptadecan-9-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(2-(4-propylcyclohexyl)ethoxy)octyl )amino)octanoate [00468] Compound 39 was prepared analogously to compound 26 starting from 4- propylcyclohexanone instead of 7-tridecanone. UPLC/ELSD: RT = 2.97 min. MS (ES): m/z (MH + ) 859.727 for C 5 2H95N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (p, 1H); 4.12 (t, 2H); 3.68 (bm, 2H); 3.28 (m, 3H); 2.66-2.40 (m, 6H); 2.32 (m, 4H); 1.84-1.12 (m, 65H); 0.90 (m, 12H). BF. Compound 40: 2-(3-Ethylcyclopentyl)ethyl 8-((8-(heptadecan-9-yloxy)-8- oxooctyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)amino)octanoate [00469] Compound 40 was prepared analogously to compound 26 starting from 3- ethylcyclopentanone instead of 7-tridecanone. UPLC/ELSD: RT = 2.84 min. MS (ES): m/z (MH + ) 831.604 for C 5 0H91N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.79 (p, 1H); 4.87 (t, 2H); 4.08 (bm, 2H); 3.66 (m, 3H); 3.50 (m, 2H); 3.28 (m, 3H); 2.57 (m, 2H); 2.44 (m, 4H); 2.31 (m, 4H); 2.03-1.08 (m, 63H); 0.89 (m, 8H); 0.71 (m, 1H). BG. Compound 42: Bis(3-pentyloctyl) 8,8'-((3-hydroxypropyl)azanediyl)dioctanoate [00470] To a solution of 3-pentyloctyl 8-bromooctanoate (398 mg, 0.98 mmol) and 3- pentyloctyl 8-((3-hydroxypropyl)amino)octanoate (374 mg, 0.94 mmol) in cyclopentyl methyl ether (5 mL) and acetonitrile (5 mL) was added potassium carbonate (776 mg, 5.62 mmol) and potassium iodide (171 mg, 1.03 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-pentyloctyl) 8,8'-((3- hydroxypropyl)azanediyl)dioctanoate (296 mg, 0.41 mmol, 44%) as a clear viscous oil. UPLC/ELSD: RT = 2.94 min. MS (ES): m/z (MH + ) 724.42 for C 45 H 89 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.69 (br. s, 1H); 4.08 (t, 4H, J = 6.0 Hz); 3.79 (br. t, 2H, J = 6.0 Hz); 2.63 (br. t, 2H, J = 6.0 Hz); 2.39 (br. dd, 4H, J = 9.0, 9.0 Hz); 2.28 (t, 4H, J = 6.0 Hz); 1.72- 1.52 (m, 10H); 1.51-1.20 (m, 50H); 0.88 (t, 12H, J = 6.0 Hz). BH. Compound 44: Bis(3-pentyloctyl) 8,8'-((4-hydroxybutyl)azanediyl)dioctanoate [00471] To a solution of 3-pentyloctyl 8-bromooctanoate (516 mg, 1.27 mmol) and 3- pentyloctyl 8-((4-hydroxybutyl)amino)octanoate (501 mg, 1.21 mmol) in cyclopentyl methyl ether (6 mL) and acetonitrile (6 mL) was added potassium carbonate (1.00 g, 7.27 mmol) and potassium iodide (221 mg, 1.33 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-pentyloctyl) 8,8'-((4- hydroxybutyl)azanediyl)dioctanoate (515 mg, 0.70 mmol, 58%) as a clear viscous oil. UPLC/ELSD: RT = 2.93 min. MS (ES): m/z (MH + ) 738.35 for C 46 H 91 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 6.56 (br. s, 1H); 4.08 (t, 4H, J = 6.0 Hz); 3.54 (br. t, 2H, J = 6.0 Hz); 2.42 (br. t, 6H, J = 6.0 Hz); 2.28 (t, 4H, J = 6.0 Hz); 1.71-1.52 (m, 12H); 1.52-1.19 (m, 50H); 0.88 (t, 12H, J = 6.0 Hz). BI. Compound 49: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(3- hydroxypropyl)amino)octanoate [00472] To a solution of 3-butylheptyl 8-bromooctanoate (629 mg, 1.67 mmol) and heptadecan-9-yl 8-((3-hydroxypropyl)amino)octanoate (723 mg, 1.59 mmol) in cyclopentyl methyl ether (7 mL) and acetonitrile (7 mL) was added potassium carbonate (1.32 g, 9.52 mmol) and potassium iodide (290 mg, 1.75 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((8-(heptadecan-9-yloxy)-8- oxooctyl)(3-hydroxypropyl)amino)octanoate (503 mg, 0.67 mmol, 42%) as a clear viscous oil. UPLC/ELSD: RT = 3.04 min. MS (ES): m/z (MH + ) 752.42 for C 47 H 93 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 5.68 (br. s, 1H); 4.86 (pent., 1H, J = 6.0 Hz); 4.08 (t, 2H, J = 6.0 Hz); 3.79 (br. t, 2H, J = 6.0 Hz); 2.63 (br. t, 2H, J = 6.0 Hz); 2.39 (br. t, 4H, J = 6.0 Hz); 2.28 (dt, 4H, J = 6.0, 3.0 Hz); 1.73-1.17 (m, 65H); 0.96-0.82 (m, 12H). BJ. Compound 50: 3-Butylheptyl 8-((8-(heptadecan-9-yloxy)-8-oxooctyl)(4- hydroxybutyl)amino)octanoate [00473] To a solution of 3-butylheptyl 8-bromooctanoate (652 mg, 1.65 mmol) and heptadecan-9-yl 8-((4-hydroxybutyl)amino)octanoate (773 mg, 1.65 mmol) in cyclopentyl methyl ether (8 mL) and acetonitrile (8 mL) was added potassium carbonate (1.36 g, 9.87 mmol) and potassium iodide (300 mg, 1.81 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((8-(heptadecan-9-yloxy)-8- oxooctyl)(4-hydroxybutyl)amino)octanoate (829 mg, 1.08 mmol, 66%) as a clear viscous oil. UPLC/ELSD: RT = 3.05 min. MS (ES): m/z (MH + ) 766.48 for C 48 H 95 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 6.58 (br. s, 1H); 4.86 (pent., 1H, J = 6.0 Hz); 4.08 (t, 2H, J = 6.0 Hz); 3.55 (br. t, 2H, J = 6.0 Hz); 2.44 (br. t, 6H, J = 6.0 Hz); 2.27 (dt, 4H, J = 6.0, 3.0 Hz); 1.70- 1.41 (m, 19H); 1.36-1.18 (m, 48H); 0.94-0.81 (m, 12H). BK. Compound 51: 3-Pentyloctyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-propylhexyl)oxy)octyl)amino)oct anoate [00474] To a solution of 3-pentyloctyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (378 mg, 0.57 mmol) in ethanol (5 mL) was added 3- methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (120 mg, 0.85 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)pr opyl)(8-oxo-8- ((3-propylhexyl)oxy)octyl)amino)octanoate (198 mg, 0.26 mmol, 45%) as light-yellow amorphous solid. UPLC/ELSD: RT = 2.54 min. MS (ES): m/z (MH + ) 776.22 for C 4 6H85N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.25 (br. s, 1H); 4.07 (t, 4H, J = 6.0 Hz); 3.65 (br. s, 2H); 3.27 (d, 3H, J = 6.0 Hz); 2.54 (br. t, 2H, J = 6.0 Hz); 2.41 (br. t, 4H, J = 6.0 Hz); 2.29 (t, 4H, J = 6.0 Hz); 1.74 (br. pent., 2H, J = 6.0 Hz); 1.67-1.52 (m, 8H); 1.49-1.18 (m, 43H); 0.88 (t, 12H, J = 6.0 Hz). BL. Compound 52: 3-Butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-propylhexyl)oxy)octyl)amino)oct anoate [00475] To a solution of 3-butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (503 mg, 0.79 mmol) in ethanol (7 mL) was added 3- methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (167 mg, 1.18 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)pr opyl)(8-oxo-8- ((3-propylhexyl)oxy)octyl)amino)octanoate (228 mg, 0.31 mmol, 39%) as light-yellow amorphous solid. UPLC/ELSD: RT = 2.43 min. MS (ES): m/z (MH + ) 748.47 for C 4 4H81N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.51 (br. s, 1H); 7.19 (br. s, 1H); 4.06 (t, 4H, J = 6.0 Hz); 3.65 (br. s, 2H); 3.28 (d, 3H, J = 6.0 Hz); 2.51 (br. t, 2H, J = 6.0 Hz); 2.39 (br. t, 4H, J = 6.0 Hz); 2.27 (t, 4H, J = 6.0 Hz); 1.75 (br. pent., 2H, J = 6.0 Hz); 1.67-1.51 (m, 8H); 1.47- 1.16 (m, 38H); 0.92-0.82 (m, 12H). BM. Compound 53: Bis(3-propylhexyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate [00476] To a solution of bis(3-propylhexyl) 8,8'-((3-aminopropyl)azanediyl)dioctanoate (499 mg, 0.82 mmol) in ethanol (8 mL) was added 3-methoxy-4-(methylamino)cyclobut-3- ene-1,2-dione (173 mg, 1.23 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give bis(3-propylhexyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino )propyl)azanediyl)dioctanoate (199 mg, 0.28 mmol, 34%) as an off-white waxy solid. UPLC/ELSD: RT = 2.25 min. MS (ES): m/z (MH + ) 720.47 for C 4 2H77N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.56 (br. s, 1H); 7.25 (br. s, 1H); 4.06 (t, 4H, J = 6.0 Hz); 3.65 (br. s, 2H); 3.28 (d, 3H, J = 6.0 Hz); 2.50 (br. t, 2H, J = 6.0 Hz); 2.38 (br. t, 4H, J = 6.0 Hz); 2.27 (t, 4H, J = 6.0 Hz); 1.75 (br. pent., 2H, J = 6.0 Hz); 1.65-1.50 (m, 8H); 1.47-1.16 (m, 34H); 0.86 (t, 12H, J = 6.0 Hz). BN. Compound 54: 3-Butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-pentyloctyl)oxy)octyl)amino)oct anoate [00477] To a solution of 3-butylheptyl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate (632 mg, 0.91 mmol) in ethanol (8 mL) was added 3- methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (192 mg, 1.36 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)pr opyl)(8-oxo-8- ((3-pentyloctyl)oxy)octyl)amino)octanoate (240 mg, 0.30 mmol, 33%) as an off-white waxy solid. UPLC/ELSD: RT = 2.67 min. MS (ES): m/z (MH + ) 804.22 for C 48 H 89 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.38 (br. s, 1H); 7.03 (br. s, 1H); 4.07 (t, 4H, J = 6.0 Hz); 3.65 (br. s, 2H, J = 6.0 Hz); 3.27 (d, 3H, J = 6.0 Hz); 2.52 (br. t, 2H, J = 6.0 Hz); 2.40 (br. t, 4H, J = 6.0 Hz); 2.28 (t, 4H, J = 6.0 Hz); 1.75 (br. pent., 2H, J = 6.0 Hz); 1.67-1.51 (m, 8H); 1.47- 1.17 (m, 46H); 0.93-0.82 (m, 12H). BO. Compound 55: Bis(3-butylheptyl) 8,8'-((3-((2-(methylamino)-3,4-dioxocyclobut-1- en-1-yl)amino)propyl)azanediyl)dioctanoate [00478] UPLC/ELSD: RT = 2.52 min. MS (ES): m/z (MH + ) 776.462 for C 4 6H85N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.10 (m, 4H); 3.67 (bm, 2H); 3.29 (m, 3H); 2.58 (m, 2H); 2.43 (m, 4H); 2.32 (t, 4H); 1.84-1.16 (m, 54H); 0.91 (m, 12H). BP. Compound 57: 3-Pentyloctyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(undecan-6-yloxy)octyl)amino)octano ate: [00479] To a solution of undecan-6-yl 8-[(3-aminopropyl)[8-oxo-8-(undecan-6- yloxy)octyl]amino]octanoate (0.870 g, 1.304 mmol, 1 equiv.) in THF (6.5 mL) and water (1.1 mL) added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (0.321 g, 2.274 mmol, 1.744 equiv.). The reaction was allowed to stir at 67 °C for 16 h. The reaction was cooled to room temperature, dried, and filtered. The organics were removed under vacuum. The residue was purified by silica gel chromatography [0-30% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3-pentyloctyl 8-[(3-{[2- (methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)[8-ox o-8-(undecan-6- yloxy)octyl]amino]octanoate (0.126 g, 12%). UPLC/ELSD: RT= 2.68 min. MS (ESI): m/z calcd for C 48 H 90 N 3 O 6 + (M+H) 804.255; found, 804.47. 1 H NMR (300 MHz, CDCl 3 ) ^: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.67 (br. s, 2H); 3.30 (d, 3H); 2.59 (br. t, 2H); 2.41 (br. t, 4H); 2.31 (t, 4H); 1.78-1.28 (m, 58H); 0.90 (t, 12H). BQ. Compound 58: Nonan-5-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-pentyloctyl)oxy)octyl)amino)oct anoate [00480] To a solution of nonan-5-yl 8-((3-aminopropyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate (718 mg, 1.08 mmol) in ethanol (10 mL) was added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (228 mg, 1.61 mmol). The reaction was allowed to stir at 67 °C for 20 h. After 20 h, the reaction was cooled to room temperature and diluted with diethyl ether. The organics were washed with brine, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give nonan-5-yl 8-((3-((2-(methylamino)-3,4- dioxocyclobut-1-en-1-yl)amino)propyl)(8-oxo-8-((3-pentylocty l)oxy)octyl)amino)octanoate (215 mg, 0.28 mmol, 26%) as an off-white waxy solid. UPLC/ELSD: RT = 2.55 min. MS (ES): m/z (MH + ) 776.34 for C 46 H 85 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 7.22 (br. s, 1H); 6.80 (br. s, 1H); 4.86 (pent., 1H, J = 6.0 Hz); 4.07 (t, 2H, J = 6.0 Hz); 3.65 (br. s, 2H); 3.27 (d, 3H, J = 6.0 Hz); 2.54 (br. t, 2H, J = 6.0 Hz); 2.41 (br. t, 4H, J = 6.0 Hz); 2.29 (t, 4H, J = 6.0 Hz); 1.74 (br. pent., 2H, J = 6.0 Hz); 1.67-1.47 (m, 10H); 1.45-1.19 (m, 41H); 0.88 (t, 12H, J = 6.0 Hz). BR. Compound 60: 3-Butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(tridecan-7-yloxy)octyl)amino)octan oate Chemical Formula: C 4 8H89N3O6 Exact Mass: 803.68 Molecular Weight: 804.26 [00481] UPLC/ELSD: RT = 2.74 min. MS (ES): m/z (MH + ) 804.220 for C 4 8H89N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.70 (bm, 2H); 3.30 (m, 3H); 2.90-2.57 (m, 5H); 2.31 (m, 4H); 1.90 (bm, 2H); 1.74-1.19 (m, 56H); 0.91 (m, 12H). BS. Compound 61: 3-Butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(undecan-6-yloxy)octyl)amino)octano ate Chemical Formula: C 4 6H85N3O6 Exact Mass: 775.64 Molecular Weight: 776.20 [00482] UPLC/ELSD: RT = 2.56 min. MS (ES): m/z (MH + ) 776.344 for C 4 6H85N3O6. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.72 (bm, 2H); 3.33 (m, 3H); 3.15-2.72 (m, 5H); 2.31 (m, 4H); 2.01 (bm, 2H); 1.77-1.18 (m, 52H); 0.91 (m, 12H). BT. Compound 62: 3-Butylheptyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-(nonan-5-yloxy)-8-oxooctyl)amino)octanoat e: Chemical Formula: C 4 4H81N3O6 Exact Mass: 747.61 Molecular Weight: 748.15 [00483] UPLC/ELSD: RT = 2.36 min. MS (ES): m/z (MH + ) 748.344 for C 44 H 81 N 3 O 6 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.68 (bm, 2H); 3.30 (m, 3H); 2.74-2.43 (m, 6H); 2.31 (m, 4H); 1.82 (bm, 2H); 1.73-1.16 (m, 47H); 0.91 (m, 12H). BU. Compound 63: Pentadecan-8-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-propylhexyl)oxy)octyl)amino)oct anoate Chemical Formula: C 4 8H89N3O6 Molecular Weight: 804.255 [00484] To a solution of pentadecan-8-yl 8-[(3-aminopropyl)({8-oxo-8-[(3- propylhexyl)oxy]octyl})amino]octanoate (3.779 g, 5.436 mmol, 1 equiv.) in THF (10 mL) and added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (1.151 g, 8.154 mmol, 1.5 equiv.) in water (0.4 mL). The reaction was allowed to stir at 67 °C for 16 h under reflux. The reaction was cooled to room temperature, dried, and filtered. The organics were removed under vacuum. The residue was purified by silica gel chromatography [0-30% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain pentadecan-8-yl 8- [(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}prop yl)({8-oxo-8-[(3- propylhexyl)oxy]octyl})amino]octanoate (0.252 g, 6%). UPLC/ELSD: RT= 2.75 min. MS (ESI): m/z calcd for C 4 8H90N3O6 + (M+H) 804.255; found, 804.34. 1 H NMR (300 MHz, CDCl 3 ) ^: ppm 9.05 (br. s, 1H); 8.37 (br. s, 1 H); 4.88 (p, 1H); 4.10 (t, 2H); 3.73 (br. d, 2H); 3.34 (d, 3H); 3.26 (m, 2H); 3.07 (br. t, 4H); 2.30 (t, 4H); 2.11 (br. t, 2H); 1.71-1.28 (m, 61H); 0.90 (t, 12H). BV. Compound 64: 3-Propylhexyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(tridecan-7-yloxy)octyl)amino)octan oate Chemical Formula: C 46 H 85 N 3 O 6 Molecular Weight: 776.20 [00485] To a solution of 3-propylhexyl 8-[(3-aminopropyl)[8-oxo-8-(tridecan-7- yloxy)octyl]amino]octanoate (0.774 g, 1.16 mmol) in tetrahydrofuran (5.8 mL) and water (1.0 mL) was added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (0.246 g, 1.74 mmol). The reaction was allowed to stir at 67 °C for 18 hours. The reaction was cooled to room temperature and diluted with dichloromethane (40 mL), then washed with saturated NaHCO3 (aq.). The organic layer was separated and washed twice more with saturated NaHCO3 (aq.). The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to 3-propylhexyl 8-[(3- {[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)[ 8-oxo-8-(tridecan-7- yloxy)octyl]amino]octanoate (0.317 g, 0.375 mmol, Yield 32.3%) as a waxy yellow tinted solid. UPLC/ELSD: RT = 2.48 min found, 776.34. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.78-3.55 (m, 2H); 3.29 (d, 3H); 2.56 (t, 2H); 2.43 (t, 4H); 2.31 (t, 4H); 1.85-1.70 (m, 2H); 1.70-1.48 (m, 11H); 1.48-1.39 (m, 4H); 1.39-1.17 (m, 36H); 0.90 (t, 12H). BW. Compound 65: 3-Propylhexyl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-(undecan-6-yloxy)octyl)amino)octano ate: Chemical Formula: C 4 4H81N3O6 Molecular Weight: 748.15 [00486] To a solution of 3-propylhexyl 8-[(3-aminopropyl)[8-oxo-8-(undecan-6- yloxy)octyl]amino]octanoate (0.682 g, 1.07 mmol) in tetrahydrofuran (5.4 mL) and water (0.9 mL) was added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (0.226 g, 1.60 mmol). The reaction was allowed to stir at 67 °C for 18 hours. The reaction was cooled to room temperature and diluted with dichloromethane (40 mL), then washed with saturated NaHCO3 (aq.). The organic layer was separated and washed twice more with saturated NaHCO3 (aq.). The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain 3-propylhexyl 8- [(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}prop yl)[8-oxo-8-(undecan-6- yloxy)octyl]amino]octanoate (0.377 g, 44.8%) as a waxy yellow/white tinted solid. UPLC/ELSD: RT = 2.42 min found, 748.47. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.76-3.57 (m, 2H); 3.29 (d, 3H); 2.56 (t, 2H); 2.43 (t, 4H); 2.31 (t, 4H); 1.84-1.71 (m, 2H); 1.71-1.48 (m, 11H); 1.48-1.40 (m, 4H); 1.40-1.17 (m, 32H); 0.91 (t, 12H). BX. Compound 66: Nonan-5-yl 8-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1- yl)amino)propyl)(8-oxo-8-((3-propylhexyl)oxy)octyl)amino)oct anoate: Chemical Formula: C 4 2H77N3O6 Molecular Weight: 720.09 [00487] To a solution of nonan-5-yl 8-[(3-aminopropyl)({8-oxo-8-[(3- propylhexyl)oxy]octyl})amino]octanoate (0.20 g, 0.33 mmol) in tetrahydrofuran (1.7 mL) and water (0.3 mL) was added 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (0.070 g, 0.49 mmol). The reaction was allowed to stir at 67 °C for 18 hours. The reaction was cooled to room temperature and diluted with dichloromethane (40 mL), then washed with saturated NaHCO3 (aq.). The organic layer was separated and washed twice more with saturated NaHCO3 (aq.). The organic layer was died over MgSO 4 , then filtered and evaporated under vacuum. The residue was purified by silica gel chromatography [0-70% (mixture of 1%NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane] to obtain nonan-5-yl 8-[(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}pr opyl)({8-oxo-8- [(3-propylhexyl)oxy]octyl})amino]octanoate (0.131 g, 51.9%) as a yellow tinted waxy solid. UPLC/ELSD: RT = 2.24 min found, 720.22. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.88 (p, 1H); 4.10 (t, 2H); 3.77-3.55 (m, 2H); 3.29 (d, 3H); 2.56 (t, 2H); 2.43 (t, 4H); 2.31 (t, 4H); 1.89-1.70 (m, 3H); 1.70-1.50 (m, 10H); 1.50-1.39 (m, 4H); 1.39-1.17 (m, 28H); 0.91 (t, 12H). BY. Compound 67: 3-Pentyloctyl 8-((2-hydroxyethyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate [00488] To a solution of 3-pentyloctyl 8-bromooctanoate (517 mg, 1.28 mmol) and 3- propylhexyl 8-((2-hydroxyethyl)amino)octanoate (400 mg, 1.21 mmol) in cyclopentyl methyl ether (6 mL) and acetonitrile (6 mL) was added potassium carbonate (1.01 g, 7.28 mmol) and potassium iodide (222 mg, 1.36 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-pentyloctyl 8-((2-hydroxyethyl)(8-oxo-8-((3- propylhexyl)oxy)octyl)amino)octanoate (310 mg, 0.47 mmol, 39%) as a clear viscous oil. UPLC/ELSD: RT = 2.58 min. MS (ES): m/z (MH + ) 654.23 for C 4 0H79NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.08 (t, 4H, J = 6.0 Hz); 3.54 (br. t, 2H, J = 6.0 Hz); 2.59 (br. t, 2H, J = 6.0 Hz); 2.46 (br. t, 4H, J = 6.0 Hz); 2.28 (t, 4H, J = 6.0 Hz); 1.68-1.52 (m, 8H); 1.50-1.37 (m, 6H); 1.35-1.19 (m, 37H); 0.88 (t, 12H, J = 6.0 Hz). BZ. Compound 68: 3-Propylhexyl 8-((2-hydroxyethyl)(8-oxo-8-(undecan-6- yloxy)octyl)amino)octanoate [00489] To a solution of undecan-6-yl 8-bromooctanoate (481 mg, 1.28 mmol) and 3- propylhexyl 8-((2-hydroxyethyl)amino)octanoate (400 mg, 1.21 mmol) in cyclopentyl methyl ether (6 mL) and acetonitrile (6 mL) was added potassium carbonate (1.01 g, 7.28 mmol) and potassium iodide (222 mg, 1.36 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to room temperature, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50-100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-propylhexyl 8-((2-hydroxyethyl)(8-oxo-8- (undecan-6-yloxy)octyl)amino)octanoate (461 mg, 0.74 mmol, 61%) as a clear viscous oil. UPLC/ELSD: RT = 2.43 min. MS (ES): m/z (MH + ) 626.24 for C 28 H 75 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (pent., 1H, J = 6.0 Hz); 4.08 (t, 2H, J = 6.0 Hz); 3.52 (t, 2H, J = 6.0 Hz); 2.57 (t, 2H, J = 6.0 Hz); 2.44 (br. t, 4H, J = 6.0 Hz); 2.28 (t, 4H, J = 6.0 Hz); 1.67- 1.38 (m, 16H); 1.37-1.18 (m, 32H); 0.88 (dt, 12H, J = 6.0, 3.0 Hz). CA. Compound 69: Bis(3-butylheptyl) 8,8'-((2-hydroxyethyl)azanediyl)dioctanoate Chemical Formula: C 40 H 79 NO 5 Exact Mass: 653.60 Molecular Weight: 654.07 [00490] UPLC/ELSD: RT = 2.66 min. MS (ES): m/z (MH + ) 654.34 for C 4 0H79NO5. 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.11 (t, 4H); 3.56 (bm, 2H); 2.69 - 2.39 (bm, 6H), 2.31 (t, 4H); 1.73 – 1.17 (m, 50H); 0.92 (m, 12H). CB. Compound 70: 3-butylheptyl 8-((2-hydroxyethyl)(8-oxo-8-((3- pentyloctyl)oxy)octyl)amino)octanoate Chemical Formula: C 40 H 79 NO 5 Exact Mass: 653.60 Molecular Weight: 654.07 [00491] UPLC/ELSD: RT = 2.63 min. MS (ES): m/z (MH + ) 654.357 for C 40 H 79 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.89 (p, 1H); 4.11 (t, 2H); 3.54 (m, 2H); 3.03 (bs, 1H); 2.59 (m, 2H); 2.45 (m, 4H); 2.30 (m, 4H); 1.74-1.18 (m, 51H); 0.91 (m, 12H). CC. Compound 71: 3-Propylhexyl 8-((2-hydroxyethyl)(8-oxo-8-(tridecan-7- yloxy)octyl)amino)octanoate [00492] To a solution of tridecan-7-yl 8-bromooctanoate (517 mg, 1.28 mmol) and 3- propylhexyl 8-((2-hydroxyethyl)amino)octanoate (400 mg, 1.21 mmol) in cyclopentyl methyl ether (6 mL) and acetonitrile (6 mL) was added potassium carbonate (1.01 g, 7.28 mmol) and potassium iodide (222 mg, 1.36 mmol). The resulting reaction mixture was allowed to stir at 80 ºC for 16 h. The reaction mixture was then cooled to rt, and the volatiles were evaporated under vacuum. The resulting residue was diluted with dichloromethane and washed with water and brine. The organic layer was separated, dried (MgSO 4 ), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0-5-10-25-50- 100% (mixture of 1% NH 4 OH, 20% MeOH in dichloromethane) in dichloromethane) to give 3-propylhexyl 8-((2-hydroxyethyl)(8-oxo-8-(tridecan-7-yloxy)octyl)amino)oc tanoate (377 mg, 0.58 mmol, 47%) as a clear viscous oil. UPLC/ELSD: RT = 2.60 min. MS (ES): m/z (MH + ) 654.23 for C 40 H 79 NO 5 . 1 H NMR (300 MHz, CDCl 3 ) d: ppm 4.87 (pent., 1H, J = 6.0 Hz); 4.08 (t, 2H, J = 6.0 Hz); 3.53 (br. t, 2H, J = 6.0 Hz); 2.58 (br. t, 2H, J = 6.0 Hz); 2.45 (br. t, 4H, J = 6.0 Hz); 2.28 (dt, 4H, J = 9.0, 3.0 Hz); 1.69-1.39 (m, 16H); 1.38-1.18 (m, 36H); 0.94-0.81 (m, 12H). Example 2: Sample formulations [00493] Lipid nanoparticles (e.g., empty LNPs or loaded LNPs) including a therapeutic and/or prophylactic can be optimized according to the selection of a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), the selection of additional lipids, the amount of each lipid in the lipid component, and the wt:wt ratio of the lipid component to the therapeutic and/or prophylactic. [00494] Lipid nanoparticles (e.g., empty LNPs or loaded LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, and a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c) were prepared. Tables 2a and 2b summarize the characteristics of the formulations. [00495] As shown in Tables 2a and 2b, the choice of compound according to Formula (1- 1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A- a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c) dramatically affects the size (e.g., diameter), polydispersity index (“PDI”), and encapsulation efficiency (“%EE”) of the compositions. Table 2a. Characteristics of nanoparticles comprising compounds of the disclosure. Table 2b. Characteristics of nanoparticles comprising compounds of the disclosure. Examples 3-7: Expression, clearance, and tolerability of sample formulations [00496] The lipids of the disclosure were developed to promote potent delivery of therapeutic agents to cells, while maintaining a short half-life (i.e., a low metabolic stability) in tissue and thus reduced lipid accumulation in the tissue upon repeat dosing. High accumulation of a lipid in a tissue could trigger toxic effects, and is thus undesirable. On the other hand, a lipid which is metabolized quickly in a tissue may not deliver enough of a therapeutic agent, such as, e.g., an mRNA, to cells to be effective. The aim of the studies outlined below is the identification of amino lipids with optimal metabolic stability which also yield high potency lipid nanoparticles (LNPs). Example 3: Expression of hEPO induced by sample formulations in mice and residual lipid levels in the liver [00497] To assess potency of expression and metabolic stability of lipids of the disclosure the hepatocyte protein expression (hEPO) following administration of a nanoparticle of the disclosure (e.g., a loaded LNP) to mice was measured. [00498] Lipid nanoparticles (LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), and an mRNA encoding hEPO were intravenously administered to CD-1 mice. The concentration of hEPO in serum was tested at 6 h after injection. The particles tested had a PDI of between about 0.1–0.4, an encapsulation efficiency of between about 82–99%, and a particle diameter of about 56–145 nm. All of the tested LNPs demonstrated effective delivery of mRNA to hepatocytes with varying amounts of lipid remaining in the livers of the mice after 24h. Table 3. Expression of hEPO induced by administration of LNPs comprising lipids of the disclosure in mice and residual lipid levels in the liver.
aThe percent of total dose calculation assumes a 25 g mouse with a 1.5 g liver b >95% equals very slow metabolism of the lipid Example 4: Expression of luciferase induced by sample formulations in mice and rats [00499] To further assess potency of expression and metabolic stability of lipids of the disclosure, expression of luciferase following administration of a nanoparticle of the disclosure comprising an mRNA containing luciferase (e.g., a loaded LNP) to rodents was measured. [00500] Lipid nanoparticles (LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), and an mRNA encoding luciferase were intravenously administered to CD-1 mice and Sprague Dawley rats. The level of expression was tested via bioluminescence in the liver ex vivo at 6 h after injection. The particles tested had a PDI of between about 0.09–0.25 and an encapsulation efficiency of between about 89– 98%. All of the tested LNPs demonstrated effective delivery of mRNA to the liver and mRNA expression in hepatocytes with varying amounts of lipid remaining in the liver tissue of the rats after 24h. Table 4a. Expression of NPI-Luciferase induced by administration of LNPs comprising lipids of the disclosure in mice and rats. Table 4b. Expression of luciferase induced by administration of LNPs comprising lipids of the disclosure in mice and residual lipid levels in the liver. Example 5: Tolerability in rats [00501] To assess tolerability of the lipids of the disclosure, expression of a reporter antibody construct induced by sample formulations was assessed in rats. [00502] Lipid nanoparticles (LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), and an mRNA encoding a reporter antibody construct were intravenously administered to Sprague Dawley rats. The level of reporter antibody construct was tested via an Enzyme-linked Immunosorbent Assay (ELISA) at 6 h after injection. For the formulations tested, high levels of the reporter antibody were measured with minimal signs of liver toxicity. Table 5. Expression of reporter antibody construct induced by administration of LNPs comprising lipids of the disclosure in rats. Example 6: Expression of hEPO induced by sample formulations in rats [00503] To further assess potency of expression and metabolic stability of lipids of the disclosure the hepatocyte protein expression (hEPO) following administration of a nanoparticle of the disclosure (e.g., a loaded LNP) to rats was measured. [00504] Lipid nanoparticles (LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), and an mRNA encoding hEPO were intravenously administered to Sprague Dawley rats. The concentration of hEPO in serum was tested at 6 h after injection. All of the tested LNPs demonstrated effective delivery of mRNA to hepatocytes with varying amounts of lipid remaining in the livers of the rats after 6h. Table 6. Expression of hEPO induced by administration of LNPs comprising lipids of the disclosure in rats. Example 7: Expression of luciferase induced by sample formulations in non-human primates [00505] To the assess potency of expression and metabolic stability of lipids of the disclosure in different species, the expression of an mRNA following administration of a nanoparticle of the disclosure containing the mRNA (e.g., a loaded LNP) to non-human primates was measured. [00506] Lipid nanoparticles (LNPs) including DSPC as a phospholipid, cholesterol as a structural lipid, PEG-1 as a PEG lipid, a compound according to Formula (1-1), (2-1), (I-a), (A), (B), (A-1), (A-2), (A-3), (IA), (IB), (B-1), (B-2), (B-3), (A-a), (A-a1), (A-a2), (A-a3), (A-b), (A-b1), (A-b2), (A-b3), (A-c), or (B-c), and an mRNA encoding luciferase were intravenously administered to cynomolgus monkeys. The level of luciferase in livers ex vivo was measured via ELISA at 6 h after injection. All of the tested LNPs demonstrated effective delivery of mRNA to hepatocytes of non-human primates. Table 7. Expression of luciferase induced by administration of LNPs comprising lipids of the disclosure in non-human primates.
Enumerated Embodiments Embodiment 1. A compound of Formula (1-A): or its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein R’ branched is: R aa , R ab , R ag , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; R 2 and R 3 are independently selected from the group consisting of H, C 1-14 alkyl, C 2 -14 alkenyl, -R*YR*”, -YR*”, and -R*OR*”, or R 2 and R 3 , together with the atom to which they are attached, form a heterocycle or carbocycle; R 4 is selected from the group consisting of hydrogen, a C 3-6 carbocycle, -(CH 2 )nQ, -(CH 2 ) n CHQR, -(CH 2 ) o C(R 12 ) 2 (CH 2 ) n-o Q, -CHQR, -CQ(R) 2 , -C(O)NQR and unsubstituted C 1-6 alkyl, where Q is selected from the group consisting of a carbocycle, heterocycle, -OR, - O(CH 2 ) n N(R) 2 , -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R) R 8 , -N(R)S(O) 2 R 8 , -O(CH 2 ) n OR, -N(R)C(=NR 9 )N(R) 2 , -N(R)C(=CHR 9 )N(R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 9 )N(R) 2 , -N(OR)C(=CHR 9 )N(R) 2 , -C(=NR 9 )N(R) 2 , -C(=NR 9 )R, - C(O)N(R)OR, -(CH 2 )nN(R) 2 , –C(R)N(R) 2 C(O)OR, NC(R)=R 11 , N(C=NR 15 )R 11 , NRC(C(O)NR 14 R 14 ’) 2 , - NRC(O)(CH 2 )pC(O)NR 14 R 14 ’, wherein A is C 6-10 aryl or a heterocycle; and each o is independently selected from the group consisting of 1, 2, 3, and 4; p is 0, 1, 2, 3, or 4; a is 1, 2, 3, or 4; and each n is independently selected from the group consisting of 1, 2, 3, 4, and 5; or X a and X b are each independently O or S; R 10 is selected from the group consisting of H, halo, -OH, R, -N(R) 2 , -CN, -N 3 , -C(O)OH, -C(O)OR, -OC(O)R, -OR, -SR, -S(O)R, -S(O)OR, -S(O) 2 OR, -NO2, -S(O) 2 N(R) 2 , -N(R)S(O) 2 R, –NH(CH 2 )t1N(R) 2 , –NH(CH 2 )p1O(CH 2 )q1N(R) 2 , –NH(CH 2 ) s1 OR, –N((CH 2 ) s1 OR) 2 , -N(R)-carbocycle, -N(R)-heterocycle, -N(R)-aryl, -N(R)-heteroaryl, -N(R)(CH 2 )t1-carbocycle, -N(R)(CH 2 )t1-heterocycle, -N(R)(CH 2 )t1-aryl, - N(R)(CH 2 ) t1 -heteroaryl, a carbocycle, a heterocycle, aryl and heteroaryl; n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; r is 0 or 1; t 1 is selected from the group consisting of 1, 2, 3, 4, and 5; p 1 is selected from the group consisting of 1, 2, 3, 4, and 5; q 1 is selected from the group consisting of 1, 2, 3, 4, and 5; and s 1 is selected from the group consisting of 1, 2, 3, 4, and 5; each R 5 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; each R 6 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; M and M’ are independently selected from the group consisting of -OC(O)O-, -C(O)O-, -OC(O)-, -OC(O)-M”-C(O)O-, -OC(O)-NR M -C(O)O-, -O-M”-O-, -C(O)N(R M )-, -N(R M )C(O)-, -OC(O)N(R M )-, -N(R M )C(O)O-, -NR M C(O)NR M -, -O-N=C(R M )-, -C(O)-, - C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR M )O-, -S(O) 2 -, -S-S-, -SO-, -OS-, S(R M ) 2 O-, -O- S(R M ) 2 -, -S(O)O-, -OS(O)-, an aryl group, and a heteroaryl group, in which M” is a bond, - (CH 2 ) z C(O)-, C 1-13 alkyl, C 2-13 alkenyl, -B(R**)-, -Si(R**) 2 -, -S(R**) 2 -, or -S(O)-, wherein z is 1, 2, 3, or 4; R 7 is selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H; R 8 is selected from the group consisting of C 3-6 carbocycle and heterocycle; R 9 is selected from the group consisting of H, CN, NO 2 , C 1-6 alkyl, -OR, -S(O) 2 R, -S(O) 2 N(R) 2 , C 2-6 alkenyl, C 3-6 carbocycle and heterocycle; R 11 is selected from the group consisting of C 3-6 carbocycle and heterocycle, wherein the C 3-6 carbocycle and heterocycle are each optionally substituted with one or more R 13 ; R 12 is selected from the group consisting of H, OH, C 1-3 alkyl, and C 2-3 alkenyl; each R 13 is independently selected from the group consisting of OH, oxo, halo, C 1-6 alkyl, C 1-6 alkoxyl, C 2-6 alkenyl, C 1-6 alkylamino, di-(C 1-6 alkyl) amino, NH 2 , C(O)NH 2 , CN, and NO2; each R 14 is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; each R 14 ’ is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; R 15 is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; each R is independently selected from the group consisting of C 1-6 alkyl, C 1-3 alkyl- aryl, C 2-3 alkenyl, and H; each R’ is independently selected from the group consisting of C1-18 alkyl, C 2 -18 alkenyl, -R*YR*”, -YR*”, (CH 2 )qOR*, and H; each R M is independently selected from the group consisting of H, C 1-6 alkyl and C 2-6 alkenyl; each R*” is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; each R* is independently selected from the group consisting of C 1-12 alkyl and C 2-12 alkenyl; each R** is independently selected from the group consisting of H, OH, C 1-12 alkyl, C 2-12 alkenyl, (CH 2 )qOR*, and (CH 2 )qOH; each Y is independently a C 3-6 carbocycle; Y a is a C 3-6 carbocycle; each X is independently selected from the group consisting of F, Cl, Br, and I; each q is independently selected from the group consisting of 1, 2, and 3; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 2. A compound of Formula (1-B): its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein R aa , R ab , R ag , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; R ba , R bb , R bg , and R bd are each independently selected from the group consisting of H, C 2-3 0 alkyl, and C 5 -20 alkenyl; R 4 is selected from the group consisting of hydrogen, a C 3-6 carbocycle, -(CH 2 ) n Q, -(CH 2 )nCHQR, -(CH 2 )oC(R 12 ) 2 (CH 2 )n-oQ, -CHQR, -CQ(R) 2 , -C(O)NQR and unsubstituted C 1-6 alkyl, where Q is selected from the group consisting of a carbocycle, heterocycle, -OR, - O(CH 2 )nN(R) 2 , -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R) R 8 , -N(R)S(O) 2 R 8 , -O(CH 2 )nOR, -N(R)C(=NR 9 )N(R) 2 , -N(R)C(=CHR 9 )N(R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 9 )N(R) 2 , -N(OR)C(=CHR 9 )N(R) 2 , -C(=NR 9 )N(R) 2 , -C(=NR 9 )R, - C(O)N(R)OR, -(CH 2 )nN(R) 2 , –C(R)N(R) 2 C(O)OR, NC(R)=R 11 , N(C=NR 15 )R 11 , NRC(C(O)NR 14 R 14 ’) 2 , -NRC(O)(CH 2 ) p C(O)NR 14 R 14 ’, wherein A is C 6-10 aryl or a heterocycle; and each o is independently selected from the group consisting of 1, 2, 3, and 4; p is 0, 1, 2, 3, or 4; a is 1, 2, 3, or 4; and each n is independently selected from the group consisting of 1, 2, 3, 4, and 5; or wherein X a and X b are each independently O or S; R 10 is selected from the group consisting of H, halo, -OH, R, -N(R) 2 , -CN, -N 3 , -C(O)OH, -C(O)OR, -OC(O)R, -OR, -SR, -S(O)R, -S(O)OR, -S(O) 2 OR, -NO2, -S(O) 2 N(R) 2 , -N(R)S(O) 2 R, –NH(CH 2 ) t1 N(R) 2 , –NH(CH 2 ) p1 O(CH 2 ) q1 N(R) 2 , –NH(CH 2 )s1OR, –N((CH 2 )s1OR) 2 , -N(R)-carbocycle, -N(R)-heterocycle, -N(R)-aryl, -N(R)-heteroaryl, -N(R)(CH 2 )t1-carbocycle, -N(R)(CH 2 )t1-heterocycle, -N(R)(CH 2 )t1-aryl, - N(R)(CH 2 )t1-heteroaryl, a carbocycle, a heterocycle, aryl and heteroaryl; n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; r is 0 or 1; t 1 is selected from the group consisting of 1, 2, 3, 4, and 5; p 1 is selected from the group consisting of 1, 2, 3, 4, and 5; q 1 is selected from the group consisting of 1, 2, 3, 4, and 5; and s 1 is selected from the group consisting of 1, 2, 3, 4, and 5; each R 5 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; each R 6 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; M and M’ are independently selected from the group consisting of -OC(O)O-, -C(O)O-, -OC(O)-, -OC(O)-M”-C(O)O-, -OC(O)-NR M -C(O)O-, -O-M”-O-, -C(O)N(R M )-, -N(R M )C(O)-, -OC(O)N(R M )-, -N(R M )C(O)O-, -NR M C(O)NR M -, -O-N=C(R M )-, -C(O)-, - C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR M )O-, -S(O) 2 -, -S-S-, -SO-, -OS-, S(R M ) 2 O-, -O- S(R M ) 2 -, -S(O)O-, -OS(O)-, an aryl group, and a heteroaryl group, in which M” is a bond, - (CH 2 )zC(O)-, C 1-13 alkyl, C 2-13 alkenyl, -B(R**)-, -Si(R**) 2 -, -S(R**) 2 -, or -S(O)-, wherein z is 1, 2, 3, or 4; R 7 is selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H; R 8 is selected from the group consisting of C 3-6 carbocycle and heterocycle; R 9 is selected from the group consisting of H, CN, NO2, C 1-6 alkyl, -OR, -S(O) 2 R, -S(O) 2 N(R) 2 , C 2-6 alkenyl, C 3-6 carbocycle and heterocycle; R 11 is selected from the group consisting of C 3-6 carbocycle and heterocycle, wherein the C 3-6 carbocycle and heterocycle are each optionally substituted with one or more R 13 ; R 12 is selected from the group consisting of H, OH, C 1-3 alkyl, and C 2-3 alkenyl; each R 13 is independently selected from the group consisting of OH, oxo, halo, C 1-6 alkyl, C 1-6 alkoxyl, C 2-6 alkenyl, C 1-6 alkylamino, di-(C 1-6 alkyl) amino, NH 2 , C(O)NH 2 , CN, and NO 2 ; each R 14 is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; each R 14 ’ is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; R 15 is independently selected from the group consisting of H, OH, C 1-6 alkyl, and C 2-3 alkenyl; each R is independently selected from the group consisting of C 1-6 alkyl, C 1-3 alkyl- aryl, C 2-3 alkenyl, and H; each R’ is independently selected from the group consisting of C 1-18 alkyl, C 2-18 alkenyl, -R*YR*”, -YR*”, (CH 2 ) q OR*, and H; each R M is independently selected from the group consisting of H, C 1-6 alkyl and C 2-6 alkenyl; each R*” is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; each R* is independently selected from the group consisting of C 1-12 alkyl and C 2-12 alkenyl; each R** is independently selected from the group consisting of H, OH, C 1-12 alkyl, C 2-12 alkenyl, (CH 2 ) q OR*, and (CH 2 ) q OH; each Y is independently a C 3-6 carbocycle; Y a is a C 3-6 carbocycle; each X is independently selected from the group consisting of F, Cl, Br, and I; each q is independently selected from the group consisting of 1, 2, and 3; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 3. The compound of any one of the preceding embodiments, wherein R 7 is H. Embodiment 4. The compound of any of the preceding embodiments, wherein M is - C(O)O- or -OC(O)-. Embodiment 5. The compound of any of the preceding embodiments, wherein M’ is - C(O)O- or -OC(O)-. Embodiment 6. A compound of Formula (A): or its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein R’ branched point of attachment; wherein R aa is H, and R ab , R ag , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl, wherein at least one of R ab , R ag , and R ad is selected from the group consisting of C 2-12 alkyl and C 2-12 alkenyl; R 2 and R 3 are each C 1-14 alkyl; R 4 is selected from the group consisting of -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, - (CH 2 ) 5 OH a 10 wherein R is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R 5 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; each R 6 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; R 7 is H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 7. A compound of Formula (B): r its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein point of attachment; wherein R aa and R ab are each H, and R ag and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl, wherein at least one of R ag and R ad is selected from the group consisting of C 2-12 alkyl and C 2-12 alkenyl; R ba , R bb , R bg , and R bd are each independently selected from the group consisting of H, C 2-3 0 alkyl, and C 5 -20 alkenyl, wherein at least one of R ba , R bb , R bg , and R bd is selected from the group consisting of C 2-30 alkyl and C 5-20 alkenyl; R 4 is selected from the group consisting of -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, - wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R 5 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; each R 6 is independently selected from the group consisting of OH, C 1-3 alkyl, C 2-3 alkenyl, and H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 8. The compound of any one of the preceding embodiments, each R 5 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H. Embodiment 9. The compound of any one of the preceding embodiments, wherein each R 6 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H. Embodiment 10. The compound of any one of the preceding embodiments, wherein R 5 and R 6 are each H. Embodiment 11. A compound of Formula (1-1): its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein wherein denotes a point of attachment; wherein R ag and R bg are each independently a C 2-12 alkyl or C 2-12 alkenyl; R 2 and R 3 are each independently selected from the group consisting of C 1-14 alkyl and C 2 -14 alkenyl; R 4 is -(CH 2 ) 2 OH, wherein denotes a point of attachment; R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R’ independently is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; and s is 2 or 3. Embodiment 12. A compound of Formula (2-1): (2-1) or its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein R’ branched is: cyclic and R’ is: and R’ b is: ; wherein denotes a point of attachment; wherein R ag and R bg are each independently a C 2-12 alkyl or C 2-12 alkenyl; R 2 and R 3 are each independently selected from the group consisting of C 1-14 alkyl and C 2 -14 alkenyl; wherein denotes a point of attachment; R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each R’ independently is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; and s is 2 or 3. Embodiment 13. The compound of any one of the preceding embodiments, having the following structure: wherein R ag is a C 2-6 alkyl. Embodiment 14. The compound of any one of the preceding embodiments, having the following structure: Embodiment 15. The compound of any one of the preceding embodiments, having the following structure: Embodiment 16. The compound of any one of the preceding embodiments, wherein R ag is a C 2-6 alkyl. Embodiment 17. The compound of any one of the preceding embodiments, wherein R bg is a C 2-6 alkyl. Embodiment 18. The compound of any one of the preceding embodiments, wherein R ag and R bg are each independently a C 2-6 alkyl. Embodiment 19. The compound of any one of the preceding embodiments, wherein R 4 is -(CH 2 ) 2 OH. Embodiment 20. The compound of any one of the preceding embodiments, wherein R 4 i Embodiment 21. The compound of any one of the preceding embodiments, wherein R’ a is R’ branched . Embodiment 22. The compound of any one of the preceding embodiments, wherein R’ a is R’ cyclic . Embodiment 23. The compound of any one of the preceding embodiments, wherein R’ b i Embodiment 24. The compound of any one of the preceding embodiments, wherein R’ b i a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein wherein denotes a point of attachment; wherein R aa , R ag and R ad are each independently selected from the group consisting of C 2-12 alkyl, and C 2-12 alkenyl; and R ab is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 2 and R 3 are each C 1-14 alkyl; wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; R 7 is H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 26. A compound of Formula (A-2): r its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein denotes a point of attachment; wherein R aa , R ab , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 2 and R 3 are each C 1-14 alkyl; wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; R 7 is H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 27. A compound of Formula (A-3): r its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein denotes a point of attachment; wherein R aa , R ag and R ab are each independently selected from the group consisting of C 2-12 alkyl, and C 2-12 alkenyl; and R ad is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 2 and R 3 are each C 1-14 alkyl; wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; R 7 is H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 28. The compound of any one of the preceding embodiments, wherein R 2 and R 3 are each C 1-14 alkyl. Embodiment 29. The compound of any one of the preceding embodiments, wherein R 2 and R 3 are each C8 alkyl. Embodiment 30. The compound of any one of the preceding embodiments, wherein one of R 2 and R 3 is C 2 alkyl and the other is C8 alkyl. Embodiment 31. A compound of any one of the preceding embodiments, having the following structure: Embodiment 32. A compound of any one of the preceding embodiments, having the following structure: Embodiment 33. A compound of Formula (B-1): its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein point of attachment; wherein R aa , R ag , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ab is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R ba , R bg , and R bd are each independently selected from the group consisting of H, C 2-30 alkyl, and C 5 -20 alkenyl; and R bb is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 34. A compound of Formula (B-2): its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein denotes a point of attachment; wherein R aa , R ab , and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R ba , R bb , and R bd are each independently selected from the group consisting of H, C 2 - 30 alkyl, and C 5-20 alkenyl; and R bg is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH or , wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 35. A compound of Formula (B-3): its N-oxide, or a salt or isomer thereof, wherein R’ a is R’ branched or R’ cyclic ; wherein wherein denotes a point of attachment; wherein R aa , R ab , and R ag are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ad is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R ba , R bb , and R bg are each independently selected from the group consisting of H, C 2-30 alkyl, and C 5 -20 alkenyl; and R bd is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH or wherein denotes a point of attachment; R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 5 and R 6 are each H; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; R’ is a C 1-12 alkyl or C 2-12 alkenyl; Y a is a C 3-6 carbocycle; R*” a is selected from the group consisting of C 1-15 alkyl and C 2-15 alkenyl; l is selected from the group consisting of 1, 2, 3, 4, and 5; s is 2 or 3; and m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13. Embodiment 36. The compound of any one of the preceding embodiments wherein R’ a is R’ cyclic . Embodiment 37. The compound of any one of the preceding embodiments, wherein s is 2 or 3. Embodiment 38. The compound of any one of the preceding embodiments, wherein Y a is cyclohexyl. Embodiment 39. The compound of any one of the preceding embodiments, wherein Embodiment 40. The compound of any one of the preceding embodiments, wherein Y a is cyclopentyl. Embodiment 41. The compound of any one of the preceding embodiments, wherein Embodiment 42. The compound of any one of the preceding embodiments, wherein R*” a is a C 2 -alkyl or C 3 -alkyl. Embodiment 43. The compound of any one of the preceding embodiments wherein R’ a is R’ branched . Embodiment 44. A compound any one of the preceding embodiments, wherein R aa , R ab and R ad are each H; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 45. A compound any one of the preceding embodiments , wherein R ba , R bg and R bd are each H, and R bb is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 46. A compound any one of the preceding embodiments, wherein R ba , R bb and R bd are each H; R bg is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 47. A compound any one of the preceding embodiments, wherein R ba , R bb and R bg are each H, and R bd is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 48. A compound of Formula (A-a): wherein R ab , R ag and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl, wherein at least one of R ab , R ag , and R ad is selected from the group consisting of C 2-12 alkyl and C 2-12 alkenyl; R 4 is selected from the group consisting of -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, and -(CH 2 ) 5 OH; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 49. A compound of Formula (A-a1): thereof, wherein R ag and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ab is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 50. A compound of Formula (A-a2): its N-oxide, or a salt or isomer thereof, wherein R ab and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 51. A compound of Formula (A-a3): or its N-oxide, or a salt or isomer wherein R ab and R ag are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ad is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH o wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; M and M’ are each independently selected from the group consisting of -C(O)O- and -OC(O)-; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 52. A compound of Formula (A-b): (A-b) or its N-oxide, or a salt or isomer thereof, wherein R ab , R ag and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl, wherein at least one of R ab , R ag , and R ad is selected from the group consisting of C 2-12 alkyl and C 2-12 alkenyl; R 4 is selected from the group consisting of -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH and - (CH 2 ) 5 OH; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 53. A compound of Formula (A-b1): or its N-oxide, or a salt or isomer , wherein R ag and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ab is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH o wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 54. A compound of Formula (A-b2): its N-oxide, or a salt or isomer thereof, wherein R ab and R ad are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH , wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 55. A compound of Formula (A-b3): or its N-oxide, or a salt or isomer wherein R ab and R ag are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl; and R ad is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R 4 is -(CH 2 ) 2 OH o wherein R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 56. A compound any one of the preceding embodiments, wherein R ag and R ad are each H; and R ab is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 57. A compound any one of the preceding embodiments, wherein R ab and R ad are each H; and R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 58. A compound any one of the preceding embodiments, wherein R ab and R ag are each H; and R ad is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl. Embodiment 59. A compound of Formula (A-c): A-c) or its N-oxide, or a salt or isomer thereof, wherein R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; and R 4 is -(CH 2 ) 2 OH o and R’ is a C 1-12 alkyl. Embodiment 60. A compound of Formula (B-c): or its N-oxide, or a salt or isomer thereof, wherein R ag is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; R bg is a C 2 alkyl, C 3 alkyl, C 4 alkyl, C 5 alkyl, or C 6 alkyl; and R 4 is -(CH 2 ) 2 OH o and R’ is a C 1-12 alkyl. Embodiment 61. The compound of any one of the preceding embodiments, wherein R 4 is -(CH 2 ) 2 OH. Embodiment 62. The compound of any one of the preceding embodiments, wherein R 4 is -(CH 2 ) 3 OH. Embodiment 63. The compound of any one of the preceding embodiments, wherein R 4 is -(CH 2 ) 4 OH. Embodiment 64. The compound of any one of the preceding embodiments, wherein R 4 Embodiment 65. The compound of any one of the preceding embodiments, wherein n2 is 2. Embodiment 66. The compound of any one of the preceding embodiments, wherein R 10 is -N(R) 2 . Embodiment 67. The compound of any one of the preceding embodiments, wherein R 10 is –NHCH 3 . Embodiment 68. The compound of any one of the preceding embodiments, wherein R 4 Embodiment 69. The compound of any one of the preceding embodiments, wherein R 4 Embodiment 70. The compound of any of the preceding embodiments, wherein M’ and M are each -C(O)O-. Embodiment 71. The compound of any of the preceding embodiments, wherein M’ is - C(O)O- and M is -OC(O)-. Embodiment 72. The compound of any of the preceding embodiments, wherein M’ is - OC(O)- and M is -C(O)O-. Embodiment 73. The compound of any one of the preceding embodiments, wherein l is 5. Embodiment 74. The compound of any one of the preceding embodiments, wherein m is 7. Embodiment 75. The compound of any one of the preceding embodiments, wherein R’ is a C 1-12 alkyl or C 2-12 alkenyl. Embodiment 76. The compound of any one of the preceding embodiments, wherein R’ is a C 2 alkyl. Embodiment 77. The compound of any one of the preceding embodiments, wherein R’ is a C 3 or C 4 alkyl. Embodiment 78. The compound of any one of the preceding embodiments, wherein R’ is a C 3 alkyl. Embodiment 79. The compound of any one of the preceding embodiments, wherein R’ is a C 5 alkyl. Embodiment 80. The compound of any one of the preceding embodiments, wherein R ag is a C 2-6 alkyl and R’ is a C 3 alkyl. Embodiment 81. The compound of any one of the preceding embodiments, wherein R ag is a C 2-6 alkyl and R’ is a C 4 alkyl. Embodiment 82. The compound of any one of the preceding embodiments, wherein R ab is H. Embodiment 83. The compound of any one of the preceding embodiments, wherein R ab is a C 2 -C 4 alkyl. Embodiment 84. The compound of any one of the preceding embodiments, wherein R ab is n-propyl or n-butyl. Embodiment 85. The compound of any one of the preceding embodiments, wherein R ab is i-propyl, sec-butyl, or tert-butyl. Embodiment 86. The compound of any one of the preceding embodiments, wherein R ab is a C 5 -alkyl or a C 6 -alkyl. Embodiment 87. The compound of any one of the preceding embodiments, wherein R ag is H. Embodiment 88. The compound of any one of the preceding embodiments, wherein R ag is a C 2 -alkyl, C 3 -alkyl, or a C 4 alkyl. Embodiment 89. The compound of any one of the preceding embodiments, wherein R ag is n-propyl or n-butyl. Embodiment 90. The compound of any one of the preceding embodiments, wherein R ag is i-propyl, sec-butyl, or tert-butyl. Embodiment 91. The compound of any one of the preceding embodiments, wherein R ag is a C 5 -alkyl or a C 6 -alkyl. Embodiment 92. The compound of any one of the preceding embodiments, wherein R ad is H. Embodiment 93. The compound of any one of the preceding embodiments, wherein R ad is a C 2 -alkyl, C 3 -alkyl, or a C 4 alkyl. Embodiment 94. The compound of any one of the preceding embodiments, wherein R ad is n-propyl or n-butyl. Embodiment 95. The compound of any one of the preceding embodiments, wherein R ad is i-propyl, sec-butyl, or tert-butyl. Embodiment 96. The compound of any one of the preceding embodiments, wherein R ad is a C 5 -alkyl or a C 6 -alkyl. Embodiment 97. The compound of any one of the preceding embodiments, wherein R bb is H. Embodiment 98. The compound of any one of the preceding embodiments, wherein R bb is a C 2 -alkyl, C 3 -alkyl, or a C 4 alkyl. Embodiment 99. The compound of any one of the preceding embodiments, wherein R bb is n-propyl or n-butyl. Embodiment 100. The compound of any one of the preceding embodiments, wherein R bb is i-propyl, sec-butyl, or tert-butyl. Embodiment 101. The compound of any one of the preceding embodiments, wherein R bb is a C 5 -alkyl or a C 6 -alkyl. Embodiment 102. The compound of any one of the preceding embodiments, wherein R bg is H. Embodiment 103. The compound of any one of the preceding embodiments, wherein R bg is a C 2 -alkyl, C 3 -alkyl, or a C 4 alkyl. Embodiment 104. The compound of any one of the preceding embodiments, wherein R bg is n-propyl or n-butyl. Embodiment 105. The compound of any one of the preceding embodiments, wherein R bg is i-propyl, sec-butyl, or tert-butyl. Embodiment 106. The compound of any one of the preceding embodiments, wherein R bg is a C 5 -alkyl or a C 6 -alkyl. Embodiment 107. The compound of any one of the preceding embodiments, wherein R bd is H. Embodiment 108. The compound of any one of the preceding embodiments, wherein R bd is a C 2 -alkyl, C 3 -alkyl, or a C 4 alkyl. Embodiment 109. The compound of any one of the preceding embodiments, wherein R bd is n-propyl or n-butyl. Embodiment 110. The compound of any one of the preceding embodiments, wherein R bd is i-propyl, sec-butyl, or tert-butyl. Embodiment 111. A compound selected from:
Embodiment 112. A compound selected from:
Embodiment 113. A compound selected from:
Embodiment 114. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid. Embodiment 115. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, a PEG lipid, and one or more therapeutic and/or prophylactic agents. Embodiment 116. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising the compound in an amount from about 40% to about 60%. Embodiment 117. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising the phospholipid in an amount from about 0% to about 20%. Embodiment 118. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising the structural lipid in an amount from about 30% to about 50%. Embodiment 119. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising the PEG lipid in an amount from about 0% to about 5%. Embodiment 120. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising about 40 mol % to about 60 mol % of the compound of any one of the preceding embodiments, about 0 mol % to about 20 mol % phospholipid, about 30 mol % to about 50 mol % structural lipid, and about 0 mol % to about 5 mol % PEG lipid. Embodiment 121. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising about 30 mol % to about 60 mol % of the compound of any one of the preceding embodiments, about 0 mol % to about 30 mol % phospholipid, about 18.5 mol % to about 48.5 mol % structural lipid, and about 0 mol % to about 10 mol % PEG lipid. Embodiment 122. The loaded LNP of any one of the preceding embodiments, wherein the one or more therapeutic and/or prophylactic agents is a polynucleotide or a polypeptide. Embodiment 123. The loaded LNP of any one of the preceding embodiments, wherein the one or more therapeutic and/or prophylactic agents is a nucleic acid. Embodiment 124. The loaded LNP of any one of the preceding embodiments, wherein the one or more therapeutic and/or prophylactic agents is selected from the group consisting of a ribonucleic acid (RNA) and a deoxyribonucleic acid (DNA). Embodiment 125. The loaded LNP of any one of the preceding embodiments, wherein the DNA is selected from the group consisting of a double-stranded DNA, a single-stranded DNA (ssDNA), a partially double-stranded DNA, a triple stranded DNA, and a partially triple-stranded DNA. Embodiment 126. The loaded LNP of any one of the preceding embodiments, wherein the DNA is selected from the group consisting of a circular DNA, a linear DNA, and mixtures thereof. Embodiment 127. The loaded LNP of any one of the preceding embodiments, wherein the one or more therapeutic and/or prophylactic agents is selected from the group consisting of a plasmid expression vector, a viral expression vector, and mixtures thereof. Embodiment 128. The loaded LNP of any one of the preceding embodiments, wherein the one or more therapeutic and/or prophylactic agents is a RNA. Embodiment 129. The loaded LNP of any one of the preceding embodiments, wherein the RNA is selected from the group consisting of a single-stranded RNA, a double-stranded RNA (dsRNA), a partially double-stranded RNA, and mixtures thereof. Embodiment 130. The loaded LNP of any one of the preceding embodiments, wherein the RNA is selected from the group consisting of a circular RNA, a linear RNA, and mixtures thereof. Embodiment 131. The loaded LNP of any one of the preceding embodiments, wherein the RNA is selected from the group consisting of is selected from the group consisting of a short interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a RNA interference (RNAi) molecule, a microRNA (miRNA), an antagomir, an antisense RNA, a ribozyme, a Dicer-substrate RNA (dsRNA), a small hairpin RNA (shRNA), a messenger RNA (mRNA), and mixtures thereof. Embodiment 132. The loaded LNP of any one of the preceding embodiments, wherein the RNA is an mRNA. Embodiment 133. The loaded LNP of any one of the preceding embodiments, wherein the mRNA is a modified mRNA (mmRNA). Embodiment 134. The loaded LNP of any one of the preceding embodiments, wherein the mRNA incorporates a micro-RNA binding site (miR binding site). Embodiment 135. The loaded LNP of any one of the preceding embodiments, wherein the mRNA includes one or more of a stem loop, a chain terminating nucleoside, a polyA sequence, a polyadenylation signal, and/or a 5’ cap structure. Embodiment 136. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the phospholipid is selected from the group consisting of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphochol ine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine,1,2-dioleo yl-sn-glycero-3-phosphoetha nolamine (DOPE), 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), sphingomyelin, and mixtures thereof. Embodiment 137. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). Embodiment 138. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the structural lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof. Embodiment 139. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the structural lipid i or a salt thereof. Embodiment 140. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the structural lipid is cholesterol: or a salt thereof. Embodiment 141. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is selected from the group consisting of a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, and a PEG-modified dialkylglycerol, and mixtures thereof. Embodiment 142. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is selected from the group consisting of 1,2-dimyristoyl- sn-glycerol methoxypolyethylene glycol (PEG-DMG), 1,2-distearoyl-sn-glycero-3- phosphoethanolamine-N-[amino(polyethylene glycol)] (PEG-DSPE), PEG-disteryl glycerol (PEG-DSG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (PEG- DAG), PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), or PEG-l,2- dimyristyloxlpropyl-3-amine (PEG-c-DMA). Embodiment 143. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is PEG-DMG. Embodiment 144. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PL-I): or a salt thereof, wherein: R 3PL1 is –OR OPL1 ; R OPL1 is hydrogen, optionally substituted alkyl, or an oxygen protecting group; r PL1 is an integer between 1 and 100, inclusive; L 1 is optionally substituted C1-10 alkylene, wherein at least one methylene of the optionally substituted C 1-10 alkylene is independently replaced with optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, O, N(R NPL1 ), S, C(O), C(O)N(R NPL1 ), NR NPL1 C(O), - C(O)O, OC(O), OC(O)O, OC(O)N(R NPL1 ), NR NPL1 C(O)O, or NR NPL1 C(O)N(R NPL1 ); D is a moiety obtained by click chemistry or a moiety cleavable under physiological conditions; m PL1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; A is of the formula: each instance of of L 2 is independently a bond or optionally substituted C 1-6 alkylene, wherein one methylene unit of the optionally substituted C 1-6 alkylene is optionally replaced with O, N(R NPL1 ), S, C(O), C(O)N(R NPL1 ), NR NPL1 C(O), C(O)O, OC(O), OC(O)O, - OC(O)N(R NPL1 ), NR NPL1 C(O)O, or NR NPL1 C(O)N(R NPL1 ); each instance of R 2SL is independently optionally substituted C 1-3 0 alkyl, optionally substituted C 1-30 alkenyl, or optionally substituted C 1-30 alkynyl; optionally wherein one or more methylene units of R 2SL are independently replaced with optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, N(R NPL1 ), O, S, C(O), C(O)N(R NPL1 ), NR NPL1 C(O), - NR NPL1 C(O)N(R NPL1 ), C(O)O, OC(O), OC(O)O, OC(O)N(R NPL1 ), NR NPL1 C(O)O, C(O)S, - SC(O), C(=NR NPL1 ), C(=NR NPL1 )N(R NPL1 ), NR NPL1 C(=NR NPL1 ), - NR NPL1 C(=NR NPL1 )N(R NPL1 ), C(S), C(S)N(R NPL1 ), NR NPL1 C(S), NR NPL1 C(S)N(R NPL1 ), S(O) , OS(O), S(O)O, OS(O)O, OS(O) 2 , S(O) 2 O, OS(O) 2 O, N(R NPL1 )S(O), S(O)N(R NPL1 ), - N(R NPL1 )S(O)N(R NPL1 ), OS(O)N(R NPL1 ), N(R NPL1 )S(O)O, S(O) 2 , N(R NPL1 )S(O) 2 , - S(O) 2 N(R NPL1 ), N(R NPL1 )S(O) 2 N(R NPL1 ), OS(O) 2 N(R NPL1 ), or N(R NPL1 )S(O) 2 O; each instance of R NPL1 is independently hydrogen, optionally substituted alkyl, or a nitrogen protecting group; Ring B is optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; and p SL is 1 or 2. Embodiment 145. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PL-I-OH): or a salt thereof. Embodiment 146. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PL-II-OH): or a salt or isomer thereof, wherein: R 3PEG is–OR O ; R O is hydrogen, C 1-6 alkyl or an oxygen protecting group; r PEG is an integer between 1 and 100; R 5PEG is C 10-40 alkyl, C 10-40 alkenyl, or C 10-40 alkynyl; and optionally one or more methylene groups of R 5PEG are independently replaced with C 3 -10 carbocyclylene, 4 to 10 membered heterocyclylene, C 6-10 arylene, 4 to 10 membered heteroarylene,, –N(R NPEG )–, –O– , –S–, –C(O)–, –C(O)N(R NPEG )–, –NR NPEG C(O)–, –NR NPEG C(O)N(R NPEG )–, –C(O)O–, – OC(O)–, –OC(O)O–, –OC(O)N(R NPEG )–, –NR NPEG C(O)O–, –C(O)S–, –SC(O)–, – C(=NR NPEG )–, –C(=NR NPEG )N(R NPEG )–, –NR NPEG C(=NR NPEG )–, – NR NPEG C(=NR NPEG )N(R NPEG )–, –C(S)–, –C(S)N(R NPEG )–, –NR NPEG C(S)–, – NR NPEG C(S)N(R NPEG )–, –S(O)–, –OS(O)–, –S(O)O–, –OS(O)O–, –OS(O) 2 –, –S(O) 2 O–, – OS(O) 2 O–, –N(R NPEG )S(O)–, –S(O)N(R NPEG )–, –N(R NPEG )S(O)N(R NPEG )–, – OS(O)N(R NPEG )–, –N(R NPEG )S(O)O–, –S(O) 2 –, –N(R NPEG )S(O) 2 –, –S(O) 2 N(R NPEG )–, – N(R NPEG )S(O) 2 N(R NPEG )–, –OS(O) 2 N(R NPEG )–, or –N(R NPEG )S(O) 2 O–; and each instance of R NPEG is independently hydrogen, C 1-6 alkyl, or a nitrogen protecting group. Embodiment 147. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein in the PEG lipid of Formula (PL-II-OH), r is an integer between 40 and 50. Embodiment 148. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein in the PEG lipid of Formula (PL-II-OH), r is 45. Embodiment 149. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein in the PEG lipid of Formula (PL-II-OH), R 5 is C 17 alkyl. Embodiment 150. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PL-II): wherein r PEG is an integer between 1 and 100. Embodiment 151. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PEG-1): Embodiment 152. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of Formula (PL-III): salt or isomer thereof, wherein s PL1 is an integer between 1 and 100. Embodiment 153. The empty LNP or loaded LNP of any one of the preceding embodiments, wherein the PEG lipid is a compound of following formula: Embodiment 154. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the structural lipid is cholesterol. Embodiment 155. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the structural lipid is cholesterol and the PEG lipid is PEG 2k -DMG. Embodiment 156. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the structural lipid is cholesterol and the PEG lipid is PEG-1. Embodiment 157. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the PEG lipid is PEG 2k -DMG. Embodiment 158. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the PEG lipid is PEG-1. Embodiment 159. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC, the structural lipid is cholesterol, and the PEG lipid is PEG 2k -DMG. Embodiment 160. An empty lipid nanoparticle (empty LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC, the structural lipid is cholesterol, and the PEG lipid is PEG-1. Embodiment 161. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the structural lipid is cholesterol, and one or more therapeutic and/or prophylactic agents. Embodiment 162. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the structural lipid is cholesterol and the PEG lipid is PEG2k-DMG, and one or more therapeutic and/or prophylactic agents. Embodiment 163. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the structural lipid is cholesterol and the PEG lipid is PEG-1, and one or more therapeutic and/or prophylactic agents. Embodiment 164. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the PEG lipid is PEG 2k -DMG, and one or more therapeutic and/or prophylactic agents. Embodiment 165. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC and the PEG lipid is PEG-1, and one or more therapeutic and/or prophylactic agents. Embodiment 166. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC, the structural lipid is cholesterol, and the PEG lipid is PEG 2k -DMG, and one or more therapeutic and/or prophylactic agents. Embodiment 167. A loaded lipid nanoparticle (loaded LNP) comprising a compound of any one of the preceding embodiments, a phospholipid, a structural lipid, and a PEG lipid, wherein the phospholipid is DSPC, the structural lipid is cholesterol, and the PEG lipid is PEG-1, and one or more therapeutic and/or prophylactic agents. Embodiment 168. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising DSPC in an amount from about 0% to about 20%. Embodiment 169. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising cholesterol in an amount from about 30% to about 50%. Embodiment 170. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising PEG2k-DMG in an amount from about 0% to about 5%. Embodiment 171. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising PEG-1 in an amount from about 0% to about 5%. Embodiment 172. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising about 40 mol % to about 60 mol % of the compound of any one of the preceding embodiments, about 0 mol % to about 20 mol % DSPC, about 30 mol % to about 50 mol % cholesterol, and about 0 mol % to about 5 mol % PEG 2k -DMG. Embodiment 173. The empty LNP or loaded LNP of any one of the preceding embodiments, comprising about 40 mol % to about 60 mol % of the compound of any one of the preceding embodiments, about 0 mol % to about 20 mol % DSPC, about 30 mol % to about 50 mol % cholesterol, and about 0 mol % to about 5 mol % PEG-1. Embodiment 174. The loaded LNP of any one of the preceding embodiments the encapsulation efficiency of the therapeutic and/or prophylactic agent is between 80% and 100%. Embodiment 175. The loaded LNP of any one of the preceding embodiments, wherein the wt/wt ratio of the lipid component to the mRNA is from about 10:1 to about 60:1. Embodiment 176. The loaded LNP of any one of the preceding embodiments, wherein the wt/wt ratio of the lipid component to the mRNA is about 20:1. Embodiment 177. The loaded LNP of any one of the preceding embodiments, wherein the N:P ratio is from about 5:1 to about 8:1. Embodiment 178. A pharmaceutical composition comprising the loaded LNP of any one of the preceding embodiments and a pharmaceutically acceptable carrier. Embodiment 179. The pharmaceutical composition of any one of the preceding embodiments, further comprising a cryoprotectant, a buffer, or a combination thereof. Embodiment 180. The pharmaceutical composition of any one of the preceding embodiments, wherein the cryoprotectant comprises sucrose. Embodiment 181. The pharmaceutical composition of any one of the preceding embodiments, wherein the cryoprotectant comprises sodium acetate. Embodiment 182. The pharmaceutical composition of any one of the preceding embodiments, wherein the cryoprotectant comprises sucrose and sodium acetate. Embodiment 183. The pharmaceutical composition of any one of the preceding embodiments, wherein the buffer is selected from the group consisting of an acetate buffer, a citrate buffer, a phosphate buffer, and a tris buffer. Embodiment 184. A method of delivering a therapeutic and/or prophylactic agent to a cell within a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 185. A method of specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 186. A method for the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 187. A method of producing a polypeptide of interest in a cell within a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 188. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the loaded LNP of any one of the preceding embodiments. Embodiment 189. Use of a loaded LNP of any one of the preceding embodiments, in the manufacture of a medicament for delivering a therapeutic and/or prophylactic agent to a cell within a subject. Embodiment 190. Use of a loaded LNP of any one of the preceding embodiments, in the manufacture of a medicament for specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject. Embodiment 191. Use of a loaded LNP of any one of the preceding embodiments, in the manufacture of a medicament for the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject. Embodiment 192. Use of a loaded LNP of any one of the preceding embodiments, in the manufacture of a medicament for producing a polypeptide of interest in a cell within a subject. Embodiment 193. Use of a loaded LNP of any one of the preceding embodiments, in the manufacture of a medicament for treating a disease or disorder in a subject in need thereof. Embodiment 194. A loaded LNP of any one of the preceding embodiments, for use in delivering a therapeutic and/or prophylactic agent to a cell within a subject, wherein the delivering comprises administering a therapeutically effective amount of the loaded LNP to the subject. Embodiment 195. A loaded LNP of any one of the preceding embodiments, for use in specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject, wherein the delivering comprises administering a therapeutically effective amount of the loaded LNP to the subject. Embodiment 196. A loaded LNP of any one of the preceding embodiments, for use in the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject, wherein the use comprises administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 197. A loaded LNP of any one of the preceding embodiments, for use in producing a polypeptide of interest in a cell within a subject, the use comprises administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 198. A loaded LNP of any one of the preceding embodiments, for use in the treatment of a disease or disorder in a subject in need thereof, wherein the treatment comprises administering a therapeutically effective amount of the loaded LNP to a subject. Embodiment 199. A method of delivering a therapeutic and/or prophylactic agent to a cell within a subject, the method comprising administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Embodiment 200. A method of specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject, the method comprising administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Embodiment 201. A method for the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject, the method comprising administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Embodiment 202. A method of producing a polypeptide of interest in a cell within a subject, the method comprising administering to the subject the loaded LNP of any one of the preceding embodiments. Embodiment 203. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of any one of the preceding embodiments. Embodiment 204. Use of a pharmaceutical composition of any one of the preceding embodiments, in the manufacture of a medicament for delivering a therapeutic and/or prophylactic agent to a cell within a subject. Embodiment 205. Use of a pharmaceutical composition of any one of the preceding embodiments, in the manufacture of a medicament for specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject. Embodiment 206. Use of a pharmaceutical composition of any one of the preceding embodiments, in the manufacture of a medicament for the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject, the method comprising administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Embodiment 207. Use of a pharmaceutical composition of any one of the preceding embodiments, in the manufacture of a medicament for producing a polypeptide of interest in a cell within a subject. Embodiment 208. Use of a pharmaceutical composition of any one of the preceding embodiments, in the manufacture of a medicament for treating a disease or disorder in a subject in need thereof. Embodiment 209. A pharmaceutical composition of any one of the preceding embodiments, for use in delivering a therapeutic and/or prophylactic agent to a cell within a subject, wherein the delivering comprises administering a therapeutically effective amount of the pharmaceutical composition to the subject. Embodiment 210. A pharmaceutical composition of any one of the preceding embodiments, for use in specifically delivering a therapeutic and/or prophylactic agent to an organ of a subject, wherein the delivering comprises administering a therapeutically effective amount of the pharmaceutical composition to the subject. Embodiment 211. A pharmaceutical composition of any one of the preceding embodiments, for use in the enhanced delivery of a therapeutic and/or prophylactic to a target tissue of a subject, wherein the use comprises administering to the subject the pharmaceutical composition of any one of the preceding embodiments. Embodiment 212. A pharmaceutical composition of any one of the preceding embodiments, for use in producing a polypeptide of interest in a cell within a subject, the use comprises administering to the subject the pharmaceutical composition of any one of the preceding embodiments Embodiment 213. A pharmaceutical composition of any one of the preceding embodiments, for use in the treatment of a disease or disorder in a subject in need thereof, wherein the treatment comprises administering a therapeutically effective amount of the pharmaceutical composition to a subject. Embodiment 214. The method, use, or loaded LNP or pharmaceutical composition for use, of any one of the preceding embodiments, wherein the organ is selected from the group consisting of liver, kidney, lung, and spleen. Embodiment 215. The method, use, or loaded LNP or pharmaceutical composition for use, of any one of the preceding embodiments, wherein the target tissue is selected from the group consisting of liver, kidney, lung, and spleen. Embodiment 216. The method or loaded LNP or pharmaceutical composition for use of any one of the preceding embodiments, wherein the administering is performed parenterally. Embodiment 217. The method or loaded LNP or pharmaceutical composition for use wherein the administering is performed intramuscularly, intradermally, subcutaneously, and/or intravenously. Embodiment 218. The use of any one of the preceding claims, wherein the medicament is for parenteral administration. Embodiment 219. The use of any one of the preceding claims, wherein the medicament is for intramuscular, intradermal, subcutaneous, and/or intravenous administration. Embodiment 220. The method, use, or loaded LNP or pharmaceutical composition for use, of any one of the preceding embodiments, wherein the subject is human.
Equivalents [00507] It is to be understood that while the present disclosure has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the present disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and alterations are within the scope of the following claims.
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