Novel ionizable lipid compounds for nucleic acid delivery

Abstract

Disclosed are novel ionizable lipid compounds, compositions comprising such ionizable lipid compounds, and their related methods of use.Nanoparticle compositions also comprise novel lipids and additional lipids, such as phospholipids, structural lipids, and PEG lipids.Nanoparticle compositions further comprise bioactive agents, such as siRNA or mRNA, and are useful for delivering the bioactive agents to subjects that require them.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 478,560, filed January 5, 2023, the disclosure of which is incorporated herein by reference in its entirety.

[0002] The present disclosure provides novel ionizable lipid compounds, compositions comprising such ionizable lipid compounds, and related methods of their use. [Background technology]

[0003] The treatment and prevention of disease with bioactive agents such as small molecule drugs, proteins, and nucleic acids, including DNA and mRNA, has the potential to revolutionize modern medicine. In particular, therapeutic, diagnostic, and / or prophylactic nucleic acids have the potential to achieve long-lasting or curative effects through the inhibition, addition, substitution, or editing of genes or RNA. However, nucleic acid delivery into cells is challenging due to the relative instability and poor cell permeability of such molecules, as well as their short expression windows and the need for frequent re-administration to the subject.

[0004] Thus, there is a need to develop compounds, compositions, and methods that improve expression profiles, stability, promote internalization, increase target affinity, and reduce the need for frequent administration of nucleic acid therapeutic and prophylactic agents. Summary of the Invention

[0005] Summary of the Invention The present disclosure provides novel compounds and compositions and methods comprising same.

[0006] In one aspect, the present disclosure provides a compound of formula (I): [ka] or a salt or isomer thereof, wherein each m is independently an integer from 4 to 13, e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13, or any subrange selected within the range of 4 to 13, e.g., 4 to 9, 6 to 8, 4 to 7, 4 to 5, 5 to 9, 6 to 13, etc.; each n is independently an integer from 1 to 3, e.g., 1, 2, or 3, or any subrange selected from the range 1 to 3, e.g., 1 to 2, 2 to 3, etc.; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is independently H, [ka] or [ka] Selected from; each M1 and M2 is independently selected from -C(O)O- and -OC(O)-, wherein at least one of M1 or M2 is -C(O)O-; Q is -O- or -NH-; and each G is -(CR4R5) k - and; wherein each k is an integer from 2 to 5, e.g., 2, 3, 4, and 5; or any subrange selected from the range of 2 to 5, e.g., 2 to 3, 3 to 4, and 2 to 5, etc.; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0007] In one aspect, compounds of Formula I can include, for example, the following compounds: [ka] (also referred to as Compound 1) [ka] (also referred to as compound 68) [ka] (also referred to as compound 50) [ka] (also referred to as compound 48) [ka] (also referred to as compound 35) [ka] (also referred to as compound 53) [ka] (also referred to as compound 54) [ka] (also referred to as compound 42) [ka] (also referred to as compound 65).

[0008] In some aspects, compounds of Formula I are, for example, [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently14 alkyl; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0009] In one aspect, the present disclosure provides a general synthetic route for the synthesis of compounds of formula (I)(j): [ka]

[0010] In some aspects, compounds of Formula I are, for example, [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; each R6 is H; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0011] In one aspect, the present disclosure provides a general synthetic route for the synthesis of compounds of formula (I)(k): [ka]

[0012] In some aspects, compounds of Formula I are, for example, [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is [ka] and; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0013] In some aspects, compounds of Formula I are, for example, [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is [ka] and; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0014] In one aspect, the present disclosure provides a general synthetic route for the synthesis of compounds of formula (I)(l)(i) and (I)(l)(ii): [ka]

[0015] In some aspects, compounds of Formula I are, for example, [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0016] In one aspect, the present disclosure provides a general synthetic route for the synthesis of compounds of formula (I)(m): [ka] During the ceremony X is Cl, Br; each m is independently an integer from 4 to 13, e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13, or any subrange selected within the range of 4 to 13, e.g., 4 to 9, 6 to 8, 4 to 7, 4 to 5, 5 to 9, 6 to 13, etc.; each n is independently an integer from 1 to 3, e.g., 1, 2, or 3, or any subrange selected from the range 1 to 3, e.g., 1 to 2, 2 to 3, etc.; Each R 1 is C1-C5 alkyl, where alkyl is linear or branched; Each R2 and R3 is independently 14 alkyl; and each G is -(CR4R5) k - and; wherein each k is an integer from 2 to 5, e.g., 2, 3, 4, and 5; or any subrange selected from the range of 2 to 5, e.g., 2 to 3, 3 to 4, and 2 to 5, etc.; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0017] In one aspect, Method F is: [ka]

[0018] In one aspect, the present disclosure provides Representative Procedure 1 for the synthesis of Compound 1 / (I)(a): [ka]

[0019] In one aspect, the present disclosure provides representative procedure 2 for the synthesis of compound 53 / (I)(f): [ka]

[0020] In one aspect, the present disclosure provides Representative Procedure 3 for the synthesis of compound 54(I)(g): [ka] Here, the synthetic route to intermediate F is as follows: [ka]

[0021] In one aspect, the present disclosure provides representative procedure 4 for the synthesis of compound 65 / (I)(i): [ka]

[0022] In another aspect, the present disclosure provides a method of delivering a payload (e.g., a therapeutic, diagnostic, and / or prophylactic nucleic acid) to a cell (e.g., a mammalian cell) by administering to a subject in need thereof a nanoparticle composition comprising (i) a compound of Formula (I) and (ii) a payload, wherein administration of a therapeutically effective amount to the subject provides a therapeutic benefit to the subject.

[0023] In another aspect, the present disclosure provides a method of producing a polypeptide of interest in a cell (e.g., a mammalian cell) by contacting the cell with a nanoparticle composition comprising (i) a compound of Formula (I) and (ii) mRNA encoding the polypeptide of interest, whereby the mRNA can be translated within the cell to produce the polypeptide.

[0024] In another aspect, the present disclosure provides a method for introducing a gene into a cell (e.g., a mammalian cell) by contacting the cell with a nanoparticle composition comprising (i) a compound of formula (I) and (ii) DNA encoding a gene of interest, thereby enabling the cell to express the introduced gene.

[0025] In another aspect, the present disclosure provides a method for reducing expression of a gene in a cell by contacting the cell with a nanoparticle composition comprising (i) a compound represented by Formula (I) and (ii) an siRNA capable of reducing expression of a gene of interest, whereby the cell reduces expression of the gene of interest.

[0026] In another aspect, the present disclosure provides a nanoparticle composition comprising (i) a compound represented by Formula (I), (ii) a phospholipid moiety, and (ii) a payload. The phospholipid moiety can be selected from phospholipids known in the art, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin. The fatty acid moiety can be selected from the non-limiting group consisting of lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, erucic acid, arachidic acid, arachidonic acid, phytanic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid.For example, in one aspect, the phospholipids are independently 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-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME16.0) The phospholipid is selected from 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), and sphingomyelin. In one aspect, the phospholipid is DOPE. In another aspect, the phospholipid is DSPC. Non-naturally occurring species, including naturally occurring species with modifications and substitutions, including branching, oxidation, cyclization, and alkynes, are also contemplated.

[0027] In another aspect, the present disclosure provides a nanoparticle composition comprising (i) a compound of Formula (I), (ii) a structured lipid, and (iii) a payload. The structured lipid may be selected from structured lipids known in the art, such as cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, and α-tocopherol.

[0028] In another aspect, the present disclosure provides a nanoparticle composition comprising (i) a compound represented by Formula (I), (ii) a PEG-lipid, and (iii) a payload. The PEG-lipid may be selected from PEG-lipids known in the art, such as PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol.

[0029] In another aspect, the present disclosure provides a nanoparticle composition comprising: (i) a compound represented by formula (I); (ii) a phospholipid moiety; (iii) a structural lipid; (iv) a PEG-lipid; (v) a payload; or any combination thereof.

[0030] In some aspects, the nanoparticle compositions of the present invention are used in conjunction with other therapeutic compounds, separate from the nanoparticles, for the treatment of the same indication in an individual. In certain cases, the nanoparticles and the therapeutic agent are delivered separately or together. If delivered together, they may or may not be in the same formulation and may or may not be delivered by the same route.

[0031] In another aspect, the present disclosure provides a method of synthesizing a compound of formula (I).

[0032] In another aspect, the present disclosure provides a method of making a nanoparticle composition comprising a lipid component comprising a compound of formula (I). [Brief explanation of the drawings]

[0033] [Figure 1] FIG. 1 shows lipids of Formula I formulated into LNPs transfected into HEK293 cells expressing a Fluc-mRNA reporter compared to transfection of naked mRNA.

[0034] [Figure 2] Figure 2 shows the in vivo expression of Fluc-mRNA reporter payload in BALB mice over a 144-hour period following subcutaneous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP156 / SM-102).

[0035] [Figure 3] Figure 3 shows a scale graph comparing the in vivo expression of Fluc-mRNA reporter payload in BALB mice with that of naked mRNA and FDA-approved LNP (LNP156 / SM-102) over a 144-hour period following subcutaneous administration of an LNP formulation containing the lipid of Formula I.

[0036] [Figure 4] Figure 4 shows the weight change in BALB mice following subcutaneous administration of an LNP formulation containing the lipid of Formula I and a Fluc-mRNA reporter payload, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP156 / SM-102).

[0037] [Figure 5] Figure 5 shows the total flux AUC values ​​for in vivo expression of Fluc-mRNA reporter payload in BALB mice following subcutaneous administration of LNP formulations containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and FDA-approved LNP (LNP156 / SM-102).

[0038] [Figure 6]Figure 6 shows a scale graph comparing the total flux AUC values ​​of in vivo expression of Fluc-mRNA reporter payload with the in vivo expression of naked mRNA and FDA-approved LNP (LNP156 / SM-102) in BALB mice following subcutaneous administration of LNP formulations containing the lipid of Formula I.

[0039] [Figure 7] Figure 7 shows the 2D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0040] [Figure 8] FIG. 8 shows a scale graph comparing 2D in vivo expression of Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I with the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0041] [Figure 9] Figure 9 shows 2D weight changes in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I and a Fluc-mRNA reporter payload, compared to in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0042] [Figure 10] FIG. 10 shows the total flux AUC values ​​for in vivo expression of the Fluc-mRNA reporter payload in BALB mice following intravenous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0043] [Figure 11] FIG. 11 shows a scale graph comparing the total flux AUC values ​​of in vivo expression of Fluc-mRNA reporter payload with the in vivo expression of naked mRNA and FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®) in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I.

[0044] [Figure 12] FIG. 12 shows the 3D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0045] [Figure 13] FIG. 13 shows a scale graph comparing 3D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, with the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0046] [Figure 14] FIG. 14 shows the 3D total flux AUC values ​​for in vivo expression of the Fluc-mRNA reporter payload in BALB mice following intravenous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0047] [Figure 15]Figure 15 shows a scale graph comparing 3D total flux AUC values ​​for in vivo expression of Fluc-mRNA reporter payload with in vivo expression of naked mRNA and FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®) in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I.

[0048] The present disclosure relates to novel ionizable lipids and lipid nanoparticle compositions comprising the novel ionizable lipids. The present disclosure also provides methods for delivering therapeutic, diagnostic, and / or prophylactic agents to cells and methods for treating diseases or disorders in subjects in need thereof. For example, a method for delivering a therapeutic, diagnostic, and / or prophylactic agent to a cell includes contacting the cell with a nanoparticle composition of the present disclosure comprising a nucleic acid (e.g., DNA or RNA), whereby the nucleic acid provides a therapeutic benefit to the subject. A method for delivering a therapeutic, diagnostic, and / or prophylactic agent to a target cell or organ may include administering to a subject a nanoparticle composition comprising one or more ionizable lipids of the present disclosure and a payload.

[0049] Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The compounds of the present disclosure contemplate all such compounds, including cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures thereof, and other mixtures thereof. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are intended to be included in the present invention.

[0050] As used herein, the term "isomer" refers to any geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer, or diastereomer of a compound. The compounds may contain one or more chiral centers and / or double bonds and thus can exist as stereoisomers, such as double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). The present disclosure encompasses any and all isomers of the compounds described herein, including stereomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers, e.g., racemates.

[0051] Those skilled in the art will recognize that reactions can be optimized to favor one isomer, new schemes can be devised to produce a single isomer, or isomeric mixtures containing any of a variety of isomeric ratios can be utilized. For example, when only two isomers are combined, mixtures containing isomeric ratios of 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 are all contemplated by the present invention. Those skilled in the art will readily appreciate that similar ratios are contemplated for more complex isomeric mixtures. When one isomer is preferred, techniques such as preparative thin-layer chromatography, preparative HPLC, preparative chiral HPLC, or preparative SFC may be used to separate the isomers.

[0052] Those skilled in the art will understand that the synthetic methods described herein utilize various protecting groups. As used herein, the term "protecting group" means that a specific functional moiety, such as O, S, or N, is temporarily blocked, so that a reaction can be selectively carried out at another reactive site of a multifunctional compound. In preferred embodiments, the protecting group reacts selectively in good yield to give a stable protected substrate for the intended reaction; the protecting group should be selectively removed in good yield by a readily available, preferably non-toxic reagent that does not attack other functional groups; the protecting group forms an easily separable derivative (more preferably, without the creation of a new stereocenter); and the protecting group has minimal additional functionality to avoid further reactive sites. As detailed herein, oxygen, sulfur, nitrogen, and carbon protecting groups may be utilized. Hydroxyl protecting groups include: methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethyl)-2-methyl ... (2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-Methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-dinitrobenzhydryl, 5-dibenzosuberyl phenyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenediaminetetraacetic acid) Dithio)pentanoate (levulinoyl dithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-Dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1 ,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). For protecting 1,2- or 1,3-diols, protecting groups include methylene acetal, ethylidene acetal, 1-t-butylethylidene ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, 1-methoxyethylidene orthoester, 1-ethoxyethylidine orthoester, 1,2-Dimethoxyethylidene orthoesters, α-methoxybenzylidene orthoesters, 1-(N,N-dimethylamino)ethylidene derivatives, α-(N,N'-dimethylamino)benzylidene derivatives, 2-oxacyclopentylidene orthoesters, di-t-butylsilylene groups (DTBS), 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivatives (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivatives (TBDS), cyclic carbonates, cyclic boronic acid esters, ethyl boronic acid, and phenyl boronic acid. Amino protecting groups include methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc). ), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BQC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-Dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate (p-nitobenzyl carbamate), p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate mate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivatives, N'-p-toluenesulfonylaminocarbonyl derivatives, N'-phenylaminothiocarbonyl derivatives, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, Cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxycarbonylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isobornyl carbanate, isobutyl carbamate methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate , 2,4,6-tri-t-butylphenylcarbamate, 4-(trimethylammonium)benzylcarbamate, 2,4,6-trimethylbenzylcarbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o- Nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivatives, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-Diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane -2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-di Benzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, Np-methoximethyleneamine Dibenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylborinic acid derivatives, N-[phenyl(pentacarbonylchromium or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidate, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridine sulfenamide (Npys), p-toluenesulfonamide (Ts), benzene benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy 4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide. While exemplary protecting groups are detailed herein, it will be understood that the invention is not intended to be limited to these protecting groups; rather, a variety of additional equivalent protecting groups can be readily identified using the above criteria and utilized in the methods of the invention. Further protecting groups are described in Protective Groups in Organic Synthesis, Third Ed. Greene,TW and Wuts, PG, Eds., John Wiley & Sons, New York: 1999, the entire contents of which are incorporated herein by reference.

[0053] It will be understood that the compounds described herein can be substituted with any number of substituents or functional moieties. In general, the term "substituted" (whether preceded by the term "optionally" or not) and the substituents contained in the formulae of the present invention refer to the replacement of hydrogen radicals in a given structure with the radical of the specified substituent. When multiple positions in any given structure can be substituted with multiple substituents selected from a specified group, the substituents can be the same or different at each position. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. Furthermore, this invention is not intended to be limited in any manner by the permissible substituents of organic compounds. Combinations of substituents and variables envisioned by this invention are preferably those that result in the formation of stable compounds useful, for example, in the treatment of infectious or proliferative diseases. As used herein, the term "stable" preferably refers to compounds that have sufficient stability to permit manufacture, maintain compound integrity for a period of time sufficient to be detected, and preferably be useful for the purposes detailed herein.

[0054] As used herein, the term "aliphatic" includes both saturated and unsaturated, straight-chain (i.e., unbranched), branched, acyclic, cyclic, or polycyclic aliphatic hydrocarbons, which are optionally substituted with one or more functional groups. As will be appreciated by those of skill in the art, "aliphatic" is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties.

[0055] As used herein, the term "alkyl" refers to a saturated, straight- or branched-chain hydrocarbon radical derived by removing one hydrogen atom from a hydrocarbon moiety containing from 1 to 20 carbon atoms. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, and dodecyl.

[0056] As used herein, the term "alkenyl" refers to a monovalent group derived by removing a hydrogen atom from a hydrocarbon moiety having at least one carbon-carbon double bond. Alkenyl groups include, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like.

[0057] As used herein, the term "alkynyl" refers to a monovalent group derived by removing a hydrogen atom from a hydrocarbon having at least one carbon-carbon triple bond. Representative alkynyl groups include ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like.

[0058] As used herein, the term "carboxylic acid" refers to a group represented by the formula -CO2H.

[0059] Alkyl, alkenyl, and cyclyl groups (eg, carbocyclyl and heterocyclyl groups) may be optionally substituted, unless otherwise specified.

[0060] As used herein, the terms "aryl" and "heteroaryl" refer to stable monocyclic or polycyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated moieties having 3 to 14 carbon atoms, each of which may be substituted or unsubstituted. Substituents include, but are not limited to, any of the aforementioned substituents.

[0061] In certain aspects of the disclosure, "aryl" refers to a monocyclic or bicyclic carbocyclic ring system having one or two aromatic rings, including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like.

[0062] In certain aspects of the present disclosure, the term "heteroaryl," as used herein, refers to an aromatic ring containing a specified number of atoms (e.g., 5- to 12-membered or 5- to 10-membered heteroaryl) composed of one or more heteroatoms (e.g., 1, 2, 3, or 4 heteroatoms) selected from N, O, and S, with the remaining ring atoms being carbon. Heteroaryl groups do not contain adjacent S and O atoms. In some embodiments, the total number of S and O atoms in a heteroaryl group is 2 or less. In some embodiments, the total number of S and O atoms in a heteroaryl group is 1 or less. Heteroaryl groups may be attached to the parent structure by a carbon or nitrogen atom, where valence permits. For example, "pyridyl" includes 2-pyridyl, 3-pyridyl, and 4-pyridyl groups, and "pyrrolyl" includes 1-pyrrolyl, 2-pyrrolyl, and 3-pyrrolyl groups. When nitrogen is present in a heteroaryl ring, it may exist in an oxidized state (i.e., N+-O-), where permitted by the nature of the adjacent atoms and groups. Furthermore, when sulfur is present in a heteroaryl ring, sulfur may be present in an oxidized state (i.e., S-O- or SO2), provided that the nature of the adjacent atoms and groups permits. Heteroaryl groups may be monocyclic or polycyclic (e.g., bicyclic, tricyclic). Any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. In some cases, heteroaryl groups are monocyclic. Examples include pyrrole, pyrazole, imidazole, triazole (e.g., 1,2,3-triazole, 1,2,4-triazole, 1,2,4-triazole), tetrazole, furan, isoxazole, oxazole, oxadiazole (e.g., 1,2,3-oxadiazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole), thiophene, isothiazole, thiazole, thiadiazole (e.g., 1,2,3-thiadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole), pyridine, pyridazine, pyrimidine, pyrazine, triazine (e.g., 1,2,4-triazine, 1,3,5-triazine) and tetrazine. In some cases, both rings of the polycyclic heteroaryl group are aromatic. Examples include indole, isoindole, indazole, benzimidazole, benzotriazole, benzofuran, benzoxazole, benzisoxazole, benzoxadiazole, benzothiophene, benzothiazole, benzisothiazole, benzothiadiazole, 1H-pyrrolo[2,3-b]pyridine, 1H-pyrazolo[3,4-b]pyridine, 3H-imidazo[4,5-b]pyridine, 3H-[1,2,3]triazolo[4,5-b]pyridine. 1H-pyrrolo[3,2-b]pyridine, 1H-pyrazolo[4,3-b]pyridine, 1H-imidazo[4,5-b]pyridine, 1H-[1,2,3]triazolo[4,5-b]pyridine, 1H-pyrrolo[2,3-c]pyridine, 1H-pyrazolo[3,4-c]pyridine, 3H-imidazo[4,5-c]pyridine, 3H-[1,2,3]triazolo[4,5-c]pyridine, 1H-pyrrolo[3,2-c]pyridine, 1H-pyrazolo[4,3-c]pyridine, 1 ...imidazo[4,5-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H-imidazo[4,5-c]pyridine, 1H Midazo[4,5-c]pyridine, 1H-[1,2,3]triazolo[4,5-c]pyridine, furo[2,3-b]pyridine, oxazolo[5,4-b]pyridine, isoxazolo[5,4-b]pyridine, [1,2,3]oxadiazolo[5,4-b]pyridine, furo[3,2-b]pyridine, oxazolo[4,5-b]pyridine, isoxazolo[4,5-b]pyridine, [1,2,3]oxadiazolo[4,5-b]pyridine, furo[2,3-c]pyridine, oxazolo[5,4-b]pyridine Soxazolo[5,4-c]pyridine, isoxazolo[5,4-c]pyridine, [1,2,3]oxadiazolo[5,4-c]pyridine, furo[3,2-c]pyridine, oxazolo[4,5-c]pyridine, isoxazolo[4,5-c]pyridine, [1,2,3]oxadiazolo[4,5-c]pyridine, thieno[2,3-b]pyridine, thiazolo[5,4-b]pyridine, isothiazolo[5,4-b]pyridine, [1,2,3]thiadiazolo[5,4-b]pyridine, Thieno[3,2-b]pyridine, thiazolo[4,5-b]pyridine, isothiazolo[4,5-b]pyridine, [1,2,3]thiadiazolo[4,5-b]pyridine, thieno[2,3-c]pyridine, thiazolo[5,4-c]pyridine, isothiazolo[5,4-c]pyridine, [1,2,3]thiadiazolo[5,4-c]pyridine, thieno[3,2-c]pyridine, thiazolo[4,5-c]pyridine, isothiazolo[4,5-c]pyridine, [1,2,3]thiadiazolo[4,5-c]pyridine, quinoline, isoquinoline, cinnoline, quinazoline, quinoxaline, Examples of suitable thiazolidins include phthalazine, naphthyridine (e.g., 1,8-naphthyridine, 1,7-naphthyridine, 1,6-naphthyridine, 1,5-naphthyridine, 2,7-naphthyridine, and 2,6-naphthyridine), imidazo[1,2-a]pyridine, 1H-pyrazolo[3,4-d]thiazole, 1H-pyrazolo[4,3-d]thiazole, and imidazo[2,1-b]thiazole.

[0063] The term "cycloalkyl," as used herein, specifically refers to groups having 3 to 7, preferably 3 to 10, carbon atoms. Suitable cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like, which, as with other aliphatic, heteroaliphatic, or heterocyclic moieties, may be optionally substituted with substituents including, but not limited to, aliphatic; heteroaliphatic; aryl; heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO; -CN; -CF; -CHCF; -CHCl; -CHOH; -CHCHOH; -CHNH; -CHSOCH; -(O)R x ;-CO2(R x );-CON(R x )2;-OC(O)R x ;-OCO2R x ;-OCON(R x )2;-N(R x )2;-S(O)2R x ;-NR x (CO)R x , where R xEach occurrence independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, where any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and herein can be substituted or unsubstituted.

[0064] As used herein, the term "heteroaliphatic" refers to an aliphatic moiety containing one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms. Heteroaliphatic moieties can be branched, unbranched, cyclic, or acyclic, and include saturated and unsaturated heterocycles such as morpholino, pyrrolidinyl, and the like. In certain embodiments, heteroaliphatic moieties are substituted by independently replacing one or more of the hydrogen atoms thereon with one or more moieties, including, but not limited to: aliphatic; heteroaliphatic; aryl; heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO; -CN; -CF; -CHCF; -CHCl; -CHOH; -CHCHOH; -CHNH; -CHSOCH; -(O)R x ;-CO2(R x );-CON(R x )2;-OC(O)R x ;-OCO2R x ;-OCON(R x )2;-N(R x )2;-S(O)2R x ;-NR x (CO)R x , where R xEach occurrence independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, where any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and herein can be substituted or unsubstituted.

[0065] As used herein, the terms "halo" and "halogen" refer to an atom selected from fluorine, chlorine, bromine, and iodine.

[0066] As used herein, the term "haloalkyl" refers to an alkyl group, as defined above, having one, two, or three halogen atoms attached thereto, and is exemplified by groups such as chloromethyl, bromoethyl, trifluoromethyl, and the like.

[0067] As used herein, the term "heterocycloalkyl" or "heterocycle" refers to a non-aromatic 5-, 6-, or 7-membered ring or polycyclic group, including bicyclic or tricyclic groups containing a fused 6-membered ring having 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, where (i) each 5-membered ring has 0 to 1 double bond and each 6-membered ring has 0 to 2 double bonds, (ii) the nitrogen and sulfur heteroatoms are optionally oxidized, (iii) the nitrogen heteroatom is optionally quaternized, and (iv) any of the above heterocycles may be fused to a benzene ring. Representative heterocycles include, but are not limited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. In certain embodiments, a "substituted heterocycloalkyl or heterocycle" group is used, which as used herein refers to a heterocycloalkyl or heterocycle group, as defined above, substituted by independently replacing one, two, or three of the hydrogen atoms thereon with, but not limited to, an aliphatic; heteroaliphatic; aryl; heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO; -CN; -CF; -CHCF; -CHCl; -CHOH; -CHCHOH; -CHNH; -CHSOCH; -(O)R x ;-CO2(R x );-CON(R x )2;-OC(O)R x ;-OCO2R x ;-OCON(R x )2;-N(R x )2;-S(O)2R x ;-NR x (CO)R x , where R xEach occurrence independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, where any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and where any of the aryl or heteroaryl substituents described above and herein can be substituted or unsubstituted.

[0068] As used herein, the terms halo and halogen refer to an atom selected from fluorine, chlorine, bromine, and iodine.

[0069] As used herein, the term "heterocyclic" refers to a non-aromatic, partially unsaturated or fully saturated, 3- to 10-membered ring system, including monocyclic rings of 3 to 8 atoms in size, and bicyclic and tricyclic ring systems that may contain an aromatic 6-membered aryl or heteroaromatic group fused to a non-aromatic ring. These heterocyclic rings include those having 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, where the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized.

[0070] The term "heteroaryl," as used herein, refers to a cyclic aromatic radical having 5 to 10 ring atoms, of which one ring atom is selected from sulfur, oxygen, and nitrogen; 0, 1, or 2 ring atoms are additional heteroatoms independently selected from sulfur, oxygen, and nitrogen; and the remaining ring atoms are carbon, wherein the radical is attached to the remainder of the molecule through any of the ring atoms, e.g., pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, etc.

[0071] As used herein, the term "squaramide" refers to a conformationally rigid cyclobutene ring composed of two carbonyl hydrogen bond acceptors adjacent to two N-H hydrogen bond donors. These cyclobutene rings contain a delocalized nitrogen lone pair, which gives the ring a four-membered structure with aromatic properties (Hückel's rule: [4n + 2] π electrons, n = 0). A description of the chemical and physical properties of squaramides can be found in LA Marchetti, LK Kumawat, N. Mao, JC Stephens, RB Elmes, "The versatility of squaramides: from supramolecular chemistry to chemical biology," Chem, 5 (2019), pp. 1398-1485, which is incorporated herein by reference in its entirety.

[0072] As used herein, a "peptide" or "protein" comprises a string of at least three amino acids linked together by peptide bonds. The terms "protein" and "peptide" may be used interchangeably. Peptide may refer to an individual peptide or a collection of peptides. The peptides of the present invention preferably contain only natural amino acids, although unnatural amino acids (i.e., compounds that do not occur in nature but can be incorporated into polypeptide chains) and / or amino acid analogs known in the art may alternatively be used. One or more of the amino acids in the peptides of the present invention may also be modified by the addition of chemical compounds such as carbohydrate groups, phosphate groups, farnesyl groups, isofarnesyl groups, fatty acid groups, and linkers for conjugation, functionalization, or other modifications. In preferred embodiments, peptide modifications result in more stable peptides (e.g., longer half-lives in vivo). These modifications may include peptide cyclization, incorporation of D-amino acids, etc. None of the modifications should substantially interfere with the desired biological activity of the peptide.

[0073] As used herein, the term "polynucleotide" or "oligonucleotide" refers to a polymer of nucleotides. The polymer may include natural nucleosides (i.e., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine), nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, C5-propynylcytidine, C5-propynyluridine, C5-bromouridine, C5-fluorouridine, C5-iodour ... C5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine), chemically modified bases, biologically modified bases (e.g., methylated bases), intercalating bases, modified sugars (e.g., 2% fluororibose, ribose, 2′-deoxyribose, arabinose, and hexose), or modified phosphate groups (e.g., phosphorothioate and 5′-N-phosphoramidite linkages).

[0074] As used herein, the terms "approximately" and "about" refer to values ​​similar to the stated reference value when applied to one or more values ​​of interest. In some aspects, the term "approximately" or "about" refers to a value within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater or less) of the stated reference value, unless otherwise stated or clear from the context (unless such value exceeds 100% of the possible value). For example, when used in the context of the amount of a given compound in the lipid component of a nanoparticle composition, "about" can mean + / - 10% of the stated value.

[0075] The term "compound" as used herein includes all isomers and isotopes of the indicated structure. "Isotopes" refer to atoms with the same atomic number but different mass numbers due to different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium. Furthermore, compounds, salts, or complexes of the present disclosure can be prepared in combination with solvents or water molecules to form solvates and hydrates by routine methods.

[0076] As used herein, the term "contacting" refers to establishing a physical connection between two or more entities. For example, contacting a cell with a nanoparticle composition means that the cell and the nanoparticle share a physical connection. Methods for contacting a cell with an external entity, both in vivo and ex vivo, are well known in the biological arts and include intravenous, intramuscular, intradermal, and subcutaneous administration methods, and may involve various amounts of the nanoparticle composition.

[0077] As used herein, the term "deliver" means to provide an entity to a destination. For example, delivering an effective amount of a bioactive agent to a subject can include administering to the subject (e.g., by intravenous, intramuscular, intradermal, or subcutaneous routes) a nanoparticle composition comprising the bioactive agent.

[0078] As used herein, "encapsulation efficiency" refers to the amount of a therapeutic and / or prophylactic agent that becomes part of a nanoparticle composition relative to the initial total amount of the therapeutic and / or prophylactic agent used to prepare the nanoparticle composition. For example, if 97 mg of a therapeutic, diagnostic, and / or prophylactic agent are encapsulated in the nanoparticle composition out of a total of 100 mg of therapeutic and / or prophylactic agent initially provided in the composition, the encapsulation efficiency can be expressed as 97%. As used herein, "encapsulation" can refer to complete, substantial, or partial enclosing, entrapment, surrounding, or enveloping.

[0079] As used herein, "expression" of a nucleic acid sequence refers to translation of mRNA into a polypeptide or protein and / or post-translational modification of a polypeptide or protein.

[0080] As used herein, the term "effective amount" of an active agent or drug delivery device refers to the amount required to induce a desired biological response.It will be understood by those skilled in the art that the effective amount of a drug or device may vary depending on factors such as the desired biological effect, the drug to be delivered, the composition of the encapsulating matrix, the target tissue, etc.For example, the effective amount of antigen-containing microparticles delivered to immunize an individual is the amount that produces an immune response sufficient to prevent infection by the organism carrying the administered antigen.

[0081] As used herein, the term "subject" or "patient" refers to any organism to which a composition according to the present disclosure can be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and / or plants.

[0082] As used herein, "administration method" can include intravenous, intramuscular, intradermal, subcutaneous, or other methods of delivering a composition to a subject. The administration method can be selected to target delivery (e.g., specifically delivery) to a particular area or system of the body.

[0083] As used herein, "modified" means non-natural. For example, RNA may be modified RNA. That is, RNA may contain one or more non-naturally occurring nucleic acid bases, nucleosides, nucleotides, or linkers.

[0084] As used herein, a "nanoparticle composition" is a composition containing one or more lipids. Nanoparticle compositions are typically on the order of micrometers or smaller in size and may contain a lipid bilayer. Nanoparticle compositions include lipid nanoparticles (LNPs), liposomes (e.g., lipid vesicles), and lipoplexes. For example, a nanoparticle composition may be a liposome having a lipid bilayer with a diameter of 500 nm or less.

[0085] As used herein, "naturally occurring" means occurring in nature without artificial modification.

[0086] As used herein, "PEG lipid" or "PEGylated lipid" refers to a lipid that includes a polyethylene glycol moiety, such as PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol.

[0087] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0088] The phrase "pharmaceutically acceptable excipient," as used herein, refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending, complexing, or dissolving an active compound) that has the properties of being substantially non-toxic and non-inflammatory in a patient. Excipients include, for example, anti-adherents, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water for hydration. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinol palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E (α-tocopherol), vitamin C, xylitol, and other species disclosed herein.

[0089] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, where the parent compound is modified by converting an existing acid or base moiety into its salt form (e.g., by reacting the free base with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxyethanesulfonate, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, such as, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Pharmaceutically acceptable salts of the present disclosure include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods.Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or in a mixture of both; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, Pharmaceutical Salts: Properties, Selection, and Use, PH Stahl and CG Wermuth (eds.), Wiley-VCH, 2008, and Berge et al., Journal of Pharmaceutical Science, 66, 1-19 (1977), each of which is incorporated herein by reference in its entirety.

[0090] As used herein, "RNA" refers to a ribonucleic acid, which may or may not occur naturally. For example, RNA may contain modified and / or non-naturally occurring components, such as one or more nucleic acid bases, nucleosides, nucleotides, or linkers. RNA may include a cap structure, a nucleoside-terminating chain, a stem-loop, a polyA sequence, and / or a polyadenylation signal. RNA may have a nucleotide sequence encoding a polypeptide of interest. For example, RNA may be messenger RNA (mRNA). Translation of an mRNA encoding a specific polypeptide, for example, in vivo translation of the mRNA in a mammalian cell, can produce the encoded polypeptide. The RNA may be selected from the non-limiting group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), mRNA, and mixtures thereof.

[0091] As used herein, the term "therapeutic agent" refers to any agent that has a therapeutic and / or diagnostic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject. The term "prophylactic agent" refers to any agent that has a prophylactic effect when administered to a subject. Therapeutic and / or prophylactic agents are also referred to as "bioactive agents" or "drugs." Such agents include, but are not limited to, small molecules, organometallic compounds, nucleic acids, proteins, peptides, polynucleotides, metals, isotopically labeled compounds, drugs, vaccines, immunological agents, and the like. As used herein, "small molecule" includes, but is not limited to, antineoplastic agents (e.g., vincristine, doxorubicin, mitoxantrone, camptothecin, cisplatin, bleomycin, cyclophosphamide, methotrexate, and streptozotocin), antitumor agents (e.g., actinomycin D, vincristine, vinblastine, cystine arabinoside, anthracyclines, alkylating agents, platinum compounds, antimetabolites, and nucleoside analogs such as methotrexate and purine and pyrimidine analogs), antiinfectives, local anesthetics (e.g., dibucaine and chlorpromazine), beta-adrenergic blockers (e.g., propranolol, timolol, and and labetolol), antihypertensives (e.g., clonidine and hydralazine), antidepressants (e.g., imipramine, amitriptyline, and doxepime), antiepileptics (e.g., phenytoin), antihistamines (e.g., diphenhydramine, chlorpheniramine, and promethazine), antibiotics / antimicrobials (e.g., gentamicin, ciprofloxacin, and cefoxitin), antifungals (e.g., miconazole, terconazole, econazole, isoconazole, butaconazole, clotrimazole, itraconazole, nystatin, naftifine, and amphotericin B), antiparasitic drugs, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, antiglaucoma drugs, vitamins, narcotics, and contrast media.

[0092] As used herein, the term "therapeutically effective amount" means an amount of an agent (e.g., nucleic acid, drug, composition, therapeutic agent, diagnostic agent, prophylactic agent, etc.) delivered that, when administered to a subject suffering from or susceptible to an infection, disease, disorder, and / or condition, is sufficient to treat, ameliorate symptoms, diagnose, prevent, and / or delay the onset of the infection, disease, disorder, and / or condition. Nanoparticle Composition

[0093] The present disclosure provides novel ionizable lipids and delivery systems based on the use of novel ionizable lipids, such as nanoparticle compositions. Described herein are nanoparticle compositions comprising a lipid component that includes a compound according to formula (I).

[0094] In one aspect, the present disclosure provides a compound represented by formula (I): [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13, e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13, or any subrange selected within the range of 4 to 13, e.g., 4 to 9, 6 to 8, 4 to 7, 4 to 5, 5 to 9, 6 to 13, etc.; each n is independently an integer from 1 to 3, e.g., 1, 2, or 3, or any subrange selected from the range 1 to 3, e.g., 1 to 2, 2 to 3, etc.; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is independently H, [ka] or [ka] Selected from; each M1 and M2 is independently selected from -C(O)O- and -OC(O)-, wherein at least one of M1 or M2 is -C(O)O-; each Q is selected from —O— or —NH—; and each G is -(CR4R5) k - and; wherein each k is an integer from 2 to 5, e.g., 2, 3, 4, and 5; or any subrange selected from the range of 2 to 5, e.g., 2 to 3, 3 to 4, and 2 to 5, etc.; and each R4 and R5 is independently selected from H and C1-C3 alkyl. Synthetic Scheme 1

[0095] In one aspect, the present disclosure provides a compound: [ka] (also referred to as compound 65) The present invention provides a method for synthesizing [ka] Synthetic Scheme 2

[0096] In one aspect, the present disclosure provides a compound: [ka] (also referred to as compound 54) The present invention provides a method for synthesizing [ka] Here, the synthetic route to intermediate F is as follows: [ka] Synthetic Scheme 3

[0097] In one aspect, the present disclosure provides a compound: [ka] (also referred to as compound 35) The present invention provides a method for synthesizing [ka] Synthetic Scheme 4

[0098] In one aspect, the present disclosure provides a compound: [ka] or a salt or isomer thereof, each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and wherein each R4 and R5 is independently selected from H and C1-C3 alkyl, and the method comprises reacting: [ka] Synthetic Scheme 5

[0099] In one aspect, the present disclosure provides a compound: [ka] or a salt or isomer thereof, each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; each R6 is H; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and wherein each R4 and R5 is independently selected from H and C1-C3 alkyl, and the method comprises reacting: [ka] Synthetic Scheme 6

[0100] In one aspect, the present disclosure provides a compound: [ka] or a salt or isomer thereof, each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is [ka] and; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and wherein each R4 and R5 is independently selected from H and C1-C3 alkyl, and the method comprises reacting: [ka] Synthetic Scheme 7

[0101] In one aspect, the present disclosure provides a compound: [ka] or a salt or isomer thereof; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; Each R6 is [ka] and; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and wherein each R4 and R5 is independently selected from H and C1-C3 alkyl, and the method comprises reacting: [ka] Synthetic Scheme 8

[0102] In one aspect, the present disclosure provides a compound: [ka] or a salt or isomer thereof, each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, and C2-C5 alkynyl, wherein the alkyl, alkenyl, or alkynyl is linear or branched; Each R2 and R3 is independently 14 alkyl; and each G is -(CR4R5) k - and; wherein each k is selected from an integer from 2 to 5; and each R4 and R5 is independently selected from H and C1-C3 alkyl, and the method comprises the following reaction: [ka] During the ceremony X is Cl, Br; each m is independently an integer from 4 to 13, e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13, or any subrange selected within the range of 4 to 13, e.g., 4 to 9, 6 to 8, 4 to 7, 4 to 5, 5 to 9, 6 to 13, etc.; each n is independently an integer from 1 to 3, e.g., 1, 2, or 3, or any subrange selected from the range 1 to 3, e.g., 1 to 2, 2 to 3, etc.; Each R 1 R1 is C1-C5 alkyl, where alkyl is linear or branched; Each R2 and R3 is independently 14 alkyl; and each G is -(CR4R5) k - and; wherein each k is an integer from 2 to 5, e.g., 2, 3, 4, and 5; or any subrange selected from the range of 2 to 5, e.g., 2 to 3, 3 to 4, and 2 to 5, etc.; and each R4 and R5 is independently selected from H and C1-C3 alkyl.

[0103] In one aspect, Method F is: [ka] Synthetic Scheme 9

[0104] In one aspect, the present disclosure provides a compound: [ka] The present invention provides a method for synthesizing [ka] Synthetic Scheme 10

[0105] In one aspect, the present disclosure provides a compound: [ka] The present invention provides a method for synthesizing [ka]

[0106] In one aspect, compounds of Formula I can include, for example, the following compounds: [ka] (also referred to as Compound 1) [ka] (also referred to as compound 68) [ka] (also referred to as compound 50) [ka] (also referred to as compound 48) [ka] (also referred to as compound 35) [ka] (also referred to as compound 53) [ka] (also referred to as compound 54) [ka] (also referred to as compound 42) [ka] (also referred to as compound 65).

[0107] In one aspect, compounds of formula I may include, for example, compounds 1-103, or salts or isomers thereof. lipid nanoparticles

[0108] In some aspects, the dimensions of the nanoparticle composition are 1 μm or less (e.g., 1 μm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm, 175 nm, 150 nm, 125 nm, 100 nm, 75 nm, 50 nm, or less), as measured, for example, by dynamic light scattering (DLS), transmission electron microscopy, scanning electron microscopy, or another method. Nanoparticle compositions include, for example, lipid nanoparticles (LNPs), liposomes, lipid vesicles, and lipoplexes. In some aspects, the nanoparticle composition is a vesicle comprising one or more lipid bilayers. In some aspects, the nanoparticle composition comprises two or more concentric bilayers separated by aqueous compartments. The lipid bilayers may be functionalized and / or crosslinked to each other. The lipid bilayer may comprise one or more ligands, proteins, or channels.

[0109] The nanoparticle composition includes a lipid component comprising at least one compound according to Formula (I). For example, the lipid component of the nanoparticle composition may include one or more of compounds (I)(a) through (I)(i). The nanoparticle composition may also include various other components. For example, the lipid component of the nanoparticle composition may include (i) a compound according to Formula (I), (ii) a phospholipid moiety, (iii) a structural lipid, (iv) a PEG-lipid, (v) a payload, or any combination thereof. The elements of the lipid component may be provided in specific fractions.

[0110] In some embodiments, the nanoparticle composition may be targeted to a particular type or class of cells (e.g., cells of a particular organ or system thereof). For example, a nanoparticle composition containing a desired bioactive agent may be specifically delivered to the liver, kidney, spleen, femur, or lung of a mammal. Specific delivery to a particular class of cells, organ, or system or group thereof means, for example, that upon administration of the nanoparticle composition to a mammal, a higher percentage of the nanoparticle composition containing the bioactive agent is delivered to the destination (e.g., tissue) of interest compared to other destinations. In some embodiments, specific delivery may result in a 2-fold, 5-fold, 10-fold, 15-fold, or greater than 20-fold increase in the amount of bioactive agent per gram of tissue in the targeted destination (e.g., tissue of interest such as the liver) compared to another destination (e.g., the spleen).

[0111] As another example of targeted or specific delivery, the bioactive agent may be mRNA encoding a protein binding partner (e.g., an antibody or functional fragment thereof, a scaffold protein, or a peptide), or a cell surface receptor may be included in the nanoparticle composition. The mRNA may additionally or alternatively be used to direct the synthesis and extracellular localization of lipids, carbohydrates, or other biological moieties. Alternatively, other payloads or components (e.g., lipids or ligands) of the nanoparticle composition may be selected based on their affinity for a specific receptor (e.g., the low-density lipoprotein receptor), allowing the nanoparticle composition to more readily interact with the target cell population containing the receptor. For example, ligands may include, but are not limited to, specific binding pair members, antibodies, monoclonal antibodies, Fv fragments, single-chain Fv (scFv) fragments, Fab' fragments, F(ab')2 fragments, single domain antibodies, camelized antibodies and their fragments, humanized antibodies and their fragments, and multivalent versions thereof; multivalent binding reagents, including monospecific or bispecific antibodies such as disulfide-stabilized Fv fragments, scFv tandems, diabodies, tribodies, or tetrabodies; and aptamers, receptors, and fusion proteins.

[0112] In some embodiments, the ligand may be a surface-bound antibody, which can allow tuning of cell targeting specificity. This is particularly useful because it allows for the production of highly specific antibodies for a desired epitope of a desired target site. In one embodiment, multiple antibodies can be expressed on the surface of a cell, each with a different specificity for a desired target. Such an approach can increase the avidity and specificity of the targeting interaction. Ionizable lipids

[0113] The nanoparticle composition may include, in addition to the lipids according to Formula (I), one or more ionizable lipids (eg, lipids that can bear a positive or partial positive charge at physiological pH).

[0114] The lipid component of the nanoparticle composition may comprise one or more phospholipid moieties, such as one or more (poly)unsaturated lipids. The phospholipids may assemble into one or more lipid bilayers. Generally, the phospholipids may comprise a phospholipid moiety and one or more fatty acid moieties.

[0115] Phospholipids useful in the compositions and methods can be selected from phospholipids known in the art, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin. The fatty acid moiety can be selected from the non-limiting group consisting of lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, erucic acid, arachidic acid, arachidonic acid, phytanic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid.For example, in some embodiments, the phospholipid is 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-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME16.0) The phospholipid is selected from the group consisting of 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), and sphingomyelin. In some embodiments, the phospholipid is DOPE. In other embodiments, the phospholipid is DSPC. Non-naturally occurring species, including naturally occurring species with modifications and substitutions, including branching, oxidation, cyclization, and alkynes, are also contemplated.

[0116] The lipid component of nanoparticle composition can comprise one or more structured lipids.Structured lipids can be selected from the structured lipids known in the art, such as cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, and α-tocopherol.In some embodiments, structured lipids comprise cholesterol and corticosteroids (such as prednisolone, dexamethasone, prednisone, and hydrocortisone), or combinations thereof.

[0117] The lipid component of nanoparticle composition may comprise one or more PEG or PEG-modified lipid.This kind of species can also be called PEGylated lipid.PEG lipid is the lipid modified with polyethylene glycol.PEG lipid can be selected from the PEG lipid known in the art, such as PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol and PEG-modified dialkylglycerol. Adjuvants

[0118] In some aspects, the nanoparticle compositions of the present disclosure may comprise one or more lipids described herein and may further comprise one or more adjuvants, which may be selected from adjuvants known in the art, such as glucopyranosyl lipid adjuvant (GLA), CpG oligodeoxynucleotides (e.g., class A or B), poly(I:C), aluminum hydroxide, and Pam3CSK4. Polynucleotides and Nucleic Acids

[0119] In some aspects, the bioactive agent delivered by the nanoparticle compositions of the present invention is a polynucleotide or nucleic acid (e.g., ribonucleic acid or deoxyribonucleic acid). Exemplary polynucleotides for use in accordance with the present disclosure include, but are not limited to, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), including messenger RNA (mRNA), hybrids thereof, RNAi inducers, RNAi agents, siRNA, shRNA, miRNA, antisense RNA, ribozymes, catalytic DNA, RNA that induces triple helix formation, aptamers, vectors, and the like. In some embodiments, the therapeutic and / or prophylactic agent is RNA. RNA useful in the compositions and methods described herein can be selected from the group consisting of, but not limited to, shortmers, antagomirs, antisense, ribozymes, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and mixtures thereof.

[0120] In one aspect, the bioactive agent is an mRNA. The mRNA can encode any polypeptide of interest, including any naturally occurring or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA can be of any size and can have any secondary structure or activity; the polypeptide can be, for example, a functional polypeptide, protein, or enzyme, and, upon expression (i.e., translation) by one or more target cells, a functional expression product (e.g., polypeptide, protein, or enzyme) is produced and, in some cases, secreted by the target cells into the subject's peripheral circulation (e.g., plasma).

[0121] In another aspect, bioactive agent is siRNA or antisense RNA.SiRNA or antisense RNA is functional in its RNA form, and can regulate, or reduce or eliminate the expression of endogenous nucleic acid or gene.In some embodiments, such encapsulated polynucleotide can be natural or recombinant, and can use either sense or antisense mechanism of action to regulate the expression of target gene or nucleic acid.For example, siRNA or antisense RNA can be selected to silence the gene associated with specific disease, disorder or pathological condition when administered to a subject in need thereof.

[0122] In some aspects, bioactive agent is shRNA or the vector or plasmid that encodes it.When suitable construct is delivered to nucleus, shRNA can be produced in its functional form in target cell and regulate the expression of endogenous nucleic acid or gene.The construct and mechanism related to shRNA are well known in the art.

[0123] Nucleic acids and polynucleotides useful in this disclosure include a first region of linked nucleosides encoding a polypeptide of interest (e.g., a coding region), a first flanking region (e.g., a 5'-UTR) located at the 5' end of the first region, a second flanking region (e.g., a 3'-UTR) located at the 3' end of the first region, at least one 5' cap region, a polyA region, one or more intron nucleotide sequences excisable from the polynucleotide, or any combination thereof. In some embodiments, the polynucleotide or nucleic acid may include a 5' cap structure, a chain-terminating nucleotide, a stem-loop, and / or a polyadenylation signal. Any one of the regions of the nucleic acid may contain one or more modified nucleosides.

[0124] The amount of bioactive agent in a nanoparticle composition can depend on the size, composition, desired target and / or use, or other characteristics of the nanoparticle composition, as well as the characteristics of the bioactive agent. For example, the amount of nucleic acid useful in a nanoparticle composition can depend on the size, sequence, and other characteristics of the nucleic acid. The relative amounts of other components (e.g., lipids) in the nanoparticle composition can also vary. In some embodiments, the weight / weight ratio of lipid component to bioactive agent in the nanoparticle composition can be about 5:1 to about 60:1, such as 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, and 60:1. The amount of bioactive agent in the nanoparticle composition can be measured using, for example, absorption spectroscopy (eg, UV-Vis spectroscopy). Pharmaceutical Composition

[0125] The nanoparticle composition can be formulated as a pharmaceutical composition. The pharmaceutical composition may contain one or more nanoparticle compositions. For example, the pharmaceutical composition can contain one or more nanoparticle compositions comprising one or more bioactive agents and a solvent, diluent, adjuvant, at least one excipient, carrier, dispersant, or combinations thereof. The pharmaceutically acceptable carrier can be selected from one or more carriers known in the art, including Tris, acetate (e.g., sodium acetate), citrate (e.g., sodium citrate), saline, PBS, or sucrose.

[0126] As used herein, a "pharmaceutically acceptable" salt, solvent, diluent, carrier, or excipient means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia for use in mammals, particularly humans. In some aspects, one or more excipients may account for more than 50% of the total mass or volume of a pharmaceutical composition, including a nanoparticle composition. For example, one or more excipients or auxiliary ingredients may account for 50%, 60%, 70%, 80%, 90%, or more of the pharmaceutical composition.

[0127] The relative amounts of one or more nanoparticle compositions, one or more pharmaceutically acceptable excipients, and / or any additional components in a pharmaceutical composition according to the present disclosure will depend on the identity, size, and / or condition of the subject being treated, and will also vary depending on the route by which the composition is administered.

[0128] In certain embodiments, pharmaceutical compositions of the present disclosure are refrigerated or frozen for storage and / or shipping (e.g., stored at 4°C or below, e.g., at about -150°C to about 0°C or at about -80°C to about -20°C, etc.). In certain embodiments, the present disclosure also relates to methods of increasing the stability of nanoparticle compositions and / or pharmaceutical compositions comprising any compound of Formula (I) by storing the nanoparticle compositions and / or pharmaceutical compositions at 4°C or below. For example, the nanoparticle compositions and / or pharmaceutical compositions disclosed herein are stable for at least about 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 1 month, at least 2 months, at least 4 months, at least 6 months, at least 8 months, at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 22 months, or at least 24 months, e.g., at temperatures of 4°C or below (e.g., about 4°C to -20°C).

[0129] Pharmaceutical compositions containing one or more nanoparticle compositions can be prepared by any method known or hereafter developed in the art of pharmacology. Generally, such preparation methods involve combining an active ingredient with an excipient and / or one or more other accessory ingredients, and then, as desired or necessary, dividing, shaping, and / or packaging the product into desired single or multiple dosage units.

[0130] Pharmaceutical compositions according to the present disclosure can be prepared, packaged, and / or sold in bulk as a single unit dose and / or as a plurality of single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition comprising a predetermined amount of an active ingredient (e.g., a nanoparticle composition). The amount of the active ingredient is generally equal to the dosage of the active ingredient administered to a subject, and / or a convenient fraction of such a dosage, for example, one-half or one-third of such a dosage.

[0131] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using appropriate dispersants, wetting agents, and / or suspending agents. Injectable preparations can be prepared in conventional forms, such as liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid prior to injection, or emulsions. Suitable excipients include, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine ​​hydrochloride, and the like. Furthermore, if necessary, injectable pharmaceutical preparations may contain small amounts of nontoxic auxiliary substances, such as wetting agents, pH buffers, and the like. Physiologically compatible buffers include, but are not limited to, Hank's solution, Ringer's solution, or physiological saline buffer. Sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any nonirritating fixed oil can be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid, can be used in the preparation of injectable preparations.

[0132] The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, and / or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0133] Pharmaceutical formulations for parenteral administration, for example, by bolus injection or continuous infusion, include aqueous solutions of the active agent in water-soluble form (e.g., formulations that may include the compound, retinoid, second lipid, stabilizer, and / or therapeutic agent). Additionally, suspensions of the active compound may be prepared as appropriate oily injection suspensions. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions. Injectable formulations may be provided in unit dosage form, for example, in ampoules or multi-dose containers with an added preservative. The formulations may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.

[0134] In addition to the above-mentioned preparation, preparation can also be formulated as depot preparation.Such long-acting preparation can be administered by intramuscular injection.Therefore, for example, preparation (for example, the preparation that can contain compound, retinoid, second lipid, stabilizer and / or therapeutic agent) can be formulated with suitable polymer or hydrophobic material (for example, as emulsion in acceptable oil) or ion exchange resin, or as poorly soluble derivative, for example as poorly soluble salt.

[0135] The compositions and formulations described can also be formulated for topical delivery, and can be applied to the skin of a subject using any suitable method for applying a topical delivery vehicle.For example, the formulation can be applied manually, by using an applicator, or by a process that includes both.After application, the formulation can be absorbed into the skin of a subject, for example, by rubbing.Application can be carried out multiple times a day, or once a day.For example, the formulation can be applied to the skin of a subject once a day, twice a day, or multiple times a day, or once every two days, once every three days, or about once a week, once every two weeks, or once every few weeks. Methods for delivering bioactive agents to cells - Patent Application 20070122997

[0136] In one aspect, the present disclosure provides a method for producing a polypeptide of interest in mammalian cells. The method for producing the polypeptide includes contacting the cells with a nanoparticle composition containing mRNA encoding the polypeptide of interest. Upon contacting the cells with the nanoparticle composition, the mRNA can be taken up into the cells and translated to produce the polypeptide of interest. The step of contacting the mammalian cells with the nanoparticle composition containing mRNA encoding the polypeptide of interest can be performed in vivo, ex vivo, in culture, or in vitro. The amount of nanoparticle composition contacted with the cells and / or the amount of mRNA therein can depend on the type of cell or tissue contacted, the means of administration, the physiochemical properties (e.g., size, charge, and chemical composition) of the nanoparticle composition and the mRNA therein, and other factors. Generally, an effective amount of the nanoparticle composition enables efficient polypeptide production in the cells. Indicators of efficiency can include polypeptide translation (as indicated by polypeptide expression), mRNA degradation levels, and immune response indicators.

[0137] In one aspect, the present disclosure provides a method for delivering a bioactive agent, such as siRNA, into cells.Suitable cells for use in the methods described herein include prokaryotes, yeast, or higher eukaryotic cells, including plant and animal cells (e.g., mammalian cells).In some aspects, the cells can be cancer cells.In other aspects, the cells can be stem cells (e.g., pHSC cell line).In some aspects, the formulations described herein can be used to transfect cells.

[0138] Contacting a cell with a nanoparticle composition containing a nucleic acid can involve or result in transfection. Phospholipids contained in the lipid component of the nanoparticle composition can facilitate transfection and / or increase transfection efficiency, for example, by interacting with and / or fusing with cellular or intracellular membranes. Method of administration to subjects

[0139] The present disclosure provides methods for delivering bioactive agents to cells or organs. While the description provided herein of nanoparticle compositions and pharmaceutical compositions comprising nanoparticle compositions primarily relates to compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to any other mammal. Modifications of compositions suitable for administration to humans to make them suitable for administration to a variety of animals are well understood, and an ordinarily skilled veterinary pharmacologist can design and / or perform such modifications with no more than routine experimentation (if any). Subjects for which administration of the compositions is contemplated include, but are not limited to, humans, other primates, and other commercially important mammals (e.g., cows, pigs, horses, sheep, cats, dogs, mice, and / or rats).

[0140] Pharmaceutical compositions containing one or more nanoparticle compositions can be administered to any patient or subject, including those who may benefit from the therapeutic effects provided by therapeutic and / or prophylactic delivery to one or more specific cells, tissues, organs, or systems or groups. While the description provided herein of nanoparticle compositions and pharmaceutical compositions containing nanoparticle compositions primarily relates to compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to any other mammal. As will be readily apparent to those skilled in the art, the therapeutically effective in vivo dosage and specific mode of administration administered to a human or non-human subject will vary depending on the age, weight, and species of mammal being treated, the particular compounds used, and the specific application for which these compounds are used.

[0141] The dosage of a pharmaceutical composition can be adjusted to achieve the desired effect and will depend on factors such as body weight, diet, concurrent medications, and other factors recognized by medical professionals. Determining effective dosage levels, i.e., the dosage levels necessary to achieve the desired result, can be accomplished by those skilled in the art using routine pharmacological methods. For example, acceptable in vitro studies can be used to establish useful doses and routes of administration of the compositions identified by the methods of the present invention using established pharmacological methods. The attending physician will know when and how to terminate, interrupt, or adjust administration due to toxicity or organ dysfunction. Conversely, the attending physician will also know to adjust treatment to higher levels if the clinical response is inadequate (precluding toxicity). Dosage and interval can be individually adjusted to provide a plasma level of the active moiety sufficient to maintain the modulating effect, or minimum effective concentration (MEC). The MEC varies from compound to compound but can be estimated from in vitro data.

[0142] In one aspect, the compositions according to the present disclosure provide a dose of about 0.0001 mg / kg to about 10 mg / kg, about 0.001 mg / kg to about 10 mg / kg, about 0.005 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.05 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 1 mg / kg to about 10 mg / kg, about 2 mg / kg to about 10 mg / kg, about 5 mg / kg to about 10 mg / kg, about 0.0001 mg / kg to about 5 mg / kg, Approximately 0.001mg / kg to approximately 5mg / kg, approximately 0.005mg / kg to approximately 5mg / kg, approximately 0.01mg / kg to approximately 5mg / kg, approximately 0.05mg / kg to approximately 5mg / kg, approximately 0.1mg / kg to approximately 5mg / kg, approximately 1mg / kg to approximately 5mg / kg, approximately 2mg / k g~about 5mg / kg, about 0.0001mg / kg~about 2.5mg / kg, about 0.001mg / kg~about 2.5mg / kg, about 0.005mg / kg~about 2.5mg / kg, about 0.01mg / kg~about 2.5mg / kg, about 0.05mg / kg~about 2.5mg / kg, Approximately 0.1mg / kg to approximately 2.5mg / kg, approximately 1mg / kg to approximately 2.5mg / kg, approximately 2mg / kg to approximately 2.5mg / kg, approximately 0.0001mg / kg to approximately 1mg / kg, approximately 0.001mg / kg to approximately 1mg / kg, approximately 0.005m g / kg~about 1mg / kg, about 0.01mg / kg~about 1mg / kg, about 0.05mg / kg~about 1mg / kg, about 0.1mg / kg~about 1mg / kg, about 0.0001mg / kg~about 0.25mg / kg, about 0.001mg / kg The nanoparticle composition may be administered at a dosage level sufficient to deliver a predetermined dose of a therapeutic and / or prophylactic agent (e.g., mRNA) of about 0.25 mg / kg, about 0.005 mg / kg to about 0.25 mg / kg, about 0.01 mg / kg to about 0.25 mg / kg, about 0.05 mg / kg to about 0.25 mg / kg, or about 0.1 mg / kg to about 0.25 mg / kg, where a 1 mg / kg (mpk) dose provides 1 mg of therapeutic and / or prophylactic agent per kg of subject body weight. In some embodiments, a nanoparticle composition of bioactive agent may be administered at a dose of about 0.001 mg / kg to about 10 mg / kg. In other aspects, a bioactive agent may be administered at a dose of about 0.005 mg / kg to about 2.5 mg / kg.In one aspect, a dose of about 0.1 mg / kg to about 1 mg / kg may be administered, while in another aspect, a dose of about 0.05 mg / kg to about 0.25 mg / kg may be administered.

[0143] The desired dosage may be delivered, for example, three times a day, twice a day, once a day, every other day, every three days, every week, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dosage may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations). In some aspects, a single dose may be administered, for example, before or after a surgical procedure or in the case of an acute disease, disorder, or condition.

[0144] Nanoparticle compositions containing one or more bioactive agents may be used in combination with one or more other bioactive agents or imaging agents. "In combination with" is not intended to imply that these agents must be administered simultaneously and / or formulated for delivery together, although these delivery methods are within the scope of this disclosure. For example, one or more nanoparticle compositions containing one or more different bioactive agents or imaging agents may be administered in combination. The compositions may be administered simultaneously with, before, or after one or more other desired therapeutic agents or medical procedures. Generally, each agent is administered at a dose and / or on a time schedule determined for that agent. In some embodiments, the present disclosure encompasses delivering the composition, or its imaging agent, bioactive composition in combination with an agent that enhances bioavailability, reduces and / or alters metabolism, inhibits excretion, and / or alters distribution in the body.

[0145] It will be further understood that bioactive or imaging active agents utilized in combination may be administered together in a single composition or separately in different compositions. Generally, it is contemplated that agents utilized in combination will be utilized at levels that do not exceed the levels utilized individually. In some embodiments, the levels utilized in combination may be lower than the levels utilized individually.

[0146] The particular combination of therapies (therapeutics or procedures) employed in a combination regimen will take into account the compatibility of the desired therapeutics and / or procedures and the desired therapeutic effect to be achieved. It will also be understood that the therapies employed may achieve a desired effect for the same disorder (e.g., administering a composition useful for treating cancer simultaneously with a chemotherapeutic agent) or may achieve a different effect (e.g., controlling any adverse effects, such as infusion-related reactions). [Example]

[0147] The ionizable lipids of the present disclosure were synthesized using commercially available firefly luciferase reporter mRNA and test lipids in fixed relative molar ratios with DSPC, cholesterol, and PEG-lipids and formulated into LNPs. Lipids dissolved in ethanol at a total concentration of 12.87 mM were mixed with the aqueous phase (pH = 4.5) containing mRNA at a 1:3 ratio by microfluidization. The total flow rate was 12 mL / min, and the N / P ratio was 6. Buffer exchange was performed using an Amicon filter. The formulation was filter-sterilized.

[0148] The transfection efficacy and cytotoxicity of LNPs were assayed in HEK293 cells and BALB mice. The LNPs of the present disclosure were found to be effectively transfected into HEK293 cells in vitro and expressed in BALB mice in vivo. This data demonstrated transfection efficacy superior to or equivalent to that of currently FDA-approved LNPs. Example 1: Transfection efficiency

[0149] The transfection efficiency of LNPs containing the novel ionizable lipid represented by Formula I was determined by detecting the expression of firefly luciferase (Fluc) in HEK293 cells. In vitro testing of LNP formulations containing the novel lipids of the present disclosure was carried out using a firefly luciferase luminescence assay.

[0150] HEK293 cells at passages 10–19, cultured in EMEM complete medium (containing MEM, 10% FBS, 1% AA, and 1% NEAA), were placed in a 96-well plate in a cell culture incubator at 37°C and 5% CO2. Equal numbers of human embryonic kidney 293 (HEK-293) cells were transfected with a commercially available LNP formulation supplemented with firefly luciferase mRNA in the presence of human apolipoprotein E3. The amount of LNP added to the cells corresponded to 0.1 μg / mL of total mRNA in the LNP sample. 10 mL of cell suspension containing 1 million cells in EMEM complete medium was plated at 10 per well. 4 A seeding volume of 1000 cells was added to each well. A positive control (e.g., RNA transfected with Lipofectamine 3000 and with known transfection efficiency), a negative control (cells treated with RNA without Lipofectamine or LNP), and three wells of untreated cells were also plated as controls. After 22 hours, 2 hours before transfection, the growth medium in all wells was replaced with 100 μL of Opti-MEM 2% FBS.

[0151] The transfection agent included a selected nucleic acid (NA) encoding the Fluc protein, Lipofectamine 3000 supplemented with P3000 reagent, or an LNP formulation. 10 μL of transfection agent was added to each well. Cells were incubated overnight in a cell culture incubator.

[0152] Twenty-four hours after transfection, cells in the 96-well plate were washed with PBS and lysed using 20 μL of 1x Cell Lysis Buffer. Luciferase assay reagent containing luciferase assay substrate dissolved in 10 mL of luciferase assay buffer was added to the wells. Luminescence from the expressed firefly luciferase was measured according to the firefly luciferase luminescence assay protocol.

[0153] FIG. 1 shows lipids of Formula I formulated into LNPs transfected into HEK293 cells expressing a Fluc-mRNA reporter compared to transfection of naked mRNA. Example 2: In vivo BALB mice, subcutaneous administration of Fluc-mRNA reporter / IVIS

[0154] LNPs containing the novel ionizable lipid of Formula I were tested in vivo using 36 female mice of the BALB / cAnNRj strain (3 mice / group / route of administration) in experiments approved by the Local Ethics Committee for Animal Experiments in Warsaw.

[0155] To examine the duration and distribution of protein production from mRNA-LNPs in vivo, 1.0 μg of luciferase mRNA-LNPs was administered subcutaneously or intravenously to mice on a q1dx1 schedule (single administration of mRNA-LNPs). Bioluminescence imaging was performed using an IVIS Spectrum CT imaging system. Mice were intraperitoneally administered with D-luciferin at a dose of 150 mg / kg. Eight minutes (subcutaneous administration) or five minutes (intravenous administration) after D-luciferin administration, mice were anesthetized with 4% isoflurane (Aerrane, Baxter) in a chamber and placed on the imaging platform with 2% isoflurane maintained via a nose cone. Mice were imaged 13 and 15 minutes (subcutaneous administration) or 10 minutes (intravenous administration) after D-luciferin administration. Bioluminescence values ​​were quantified using commercially available imaging software by measuring the photon flux (photons / second) in the region of interest where the bioluminescent signal was emitted.

[0156] Figure 2 shows the in vivo expression of Fluc-mRNA reporter payload in BALB mice over a 144-hour period following subcutaneous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP156 / SM-102).

[0157] Figure 3 shows a scale graph comparing the in vivo expression of Fluc-mRNA reporter payload in BALB mice with that of naked mRNA and FDA-approved LNP (LNP156 / SM-102) over a 144-hour period following subcutaneous administration of an LNP formulation containing the lipid of Formula I.

[0158] Figure 4 shows the weight change in BALB mice following subcutaneous administration of an LNP formulation containing the lipid of Formula I and a Fluc-mRNA reporter payload, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP156 / SM-102).

[0159] Figure 5 shows the total flux AUC values ​​for in vivo expression of Fluc-mRNA reporter payload in BALB mice following subcutaneous administration of LNP formulations containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and FDA-approved LNP (LNP156 / SM-102).

[0160] Figure 6 shows a scale graph comparing the total flux AUC values ​​of in vivo expression of Fluc-mRNA reporter payload with the in vivo expression of naked mRNA and FDA-approved LNP (LNP156 / SM-102) in BALB mice following subcutaneous administration of LNP formulations containing the lipid of Formula I. Example 3: In vivo BALB mice, intravenous administration of Fluc-mRNA reporter / IVIS (2D)

[0161] Figure 7 shows the 2D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0162] FIG. 8 shows a scale graph comparing 2D in vivo expression of Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I with the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0163] Figure 9 shows 2D weight changes in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I and a Fluc-mRNA reporter payload, compared to in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0164] FIG. 10 shows the total flux AUC values ​​for in vivo expression of the Fluc-mRNA reporter payload in BALB mice following intravenous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0165] FIG. 11 shows a scale graph comparing the total flux AUC values ​​of in vivo expression of Fluc-mRNA reporter payload with the in vivo expression of naked mRNA and FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®) in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I. Example 4: In vivo BALB mice, intravenous administration of Fluc-mRNA reporter / IVIS (3D)

[0166] FIG. 12 shows the 3D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0167] FIG. 13 shows a scale graph comparing 3D in vivo expression of a Fluc-mRNA reporter payload in BALB mice over a 144-hour period following intravenous administration of an LNP formulation containing a lipid of Formula I, with the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0168] FIG. 14 shows the 3D total flux AUC values ​​for in vivo expression of the Fluc-mRNA reporter payload in BALB mice following intravenous administration of an LNP formulation containing the lipid of Formula I, compared to the in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0169] FIG. 15 shows a scale graph comparing 3D total flux AUC values ​​of in vivo expression of Fluc-mRNA reporter payload with in vivo expression of naked mRNA and FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®) in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I.

[0170] Figure 9 shows 2D weight changes in BALB mice following intravenous administration of LNP formulations containing the lipid of Formula I and a Fluc-mRNA reporter payload, compared to in vivo expression of naked mRNA and an FDA-approved LNP (LNP167 / D-LIN-MC3 Onpattro®).

[0171] Example 5: Representative synthesis procedure

[0172] Some compounds of formula I can be synthesized using Representative Procedure 1 described below: [ka]

[0173] Some compounds of formula I can be synthesized using representative procedure 2 described below: [ka]

[0174] Some compounds of formula I can be synthesized using representative procedure 3 described below: [ka]

[0175] Some compounds of formula I can be synthesized using representative procedure 4 described below: [ka]

[0176] Pentyl 9-bromononanoate (Intermediate A) (Method A): [ka]

[0177] To crystals of 9-bromononanoic acid (15 g, 63.3 mmol, 1 equiv.) was added thionyl chloride (4.9 mL, 66.4 mmol, 1.05 equiv.) over 10 min with stirring at room temperature under an argon atmosphere. The mixture was stirred at room temperature for 1 h and then at 60 °C for 5 h. Panthan-1-ol (8.26 mL, 75.9 mmol, 1.2 equiv.) was then added to the reaction mixture over 30 min at 60 °C. The reaction mixture was then stirred at 60 °C for 20 h, at 80 °C for 7 h, and at 120 °C for 1 h. The reaction mixture was then diluted with hexane (50 mL) and subsequently washed with saturated sodium bicarbonate solution (100 mL), water (100 mL), and brine (100 mL). The organic layer was dried over sodium sulfate, and the solvent was evaporated. The residual crude product (pentyl 9-bromononanoate, intermediate A) (18.9 g, 61.5 mmol, 97%) was obtained as a thick yellow oil and used in the next reaction without further purification. 1 H NMR (600 MHz, CDCl3) δ 4.06 (t, J=6.8 Hz, 2H), 3.40 (t, J=6.8 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.85 (qu, J=7.0 Hz, 2H), 1.68-1.58 (m, 4H), 1.48-1.38 (m, 2H), 1.38-1.26 (m, 10H), 0.91 (t, J=7.0 Hz, 3H); 13 C NMR (150 MHz, CDCl3) δ 174.0, 64.5, 34.5, 34.1, 32.9, 29.2, 29.2, 28.7, 28.5, 28.2, 25.1, 22.5, 14.1.

[0178] Pentyl 9-((2-hydroxyethyl)amino)nonanoate (Intermediate B) (Method B): [ka]

[0179] A mixture of pentyl 9-bromononanoate (5.00 g, 16.3 mmol, 1.00 equiv), ethanolamine (1.98 mL, 32.5 mmol, 2.00 equiv), potassium carbonate (9.09 g, 65.1 mmol, 4 equiv), and potassium iodide (2.99 g, 17.9 mmol, 1.10 equiv) in a mixture of dioxane and acetonitrile (125 mL, 4:1, v / v) was stirred at 80 °C for 16 h. The reaction mixture was cooled, and the solid was filtered off and washed with acetonitrile (50 mL). The filtrate was evaporated and the residue was purified on silica gel (phase A: 100% DCM, phase B: DCM / MeOH / NHOH, 80:20:1, v / v / v; phase A:B, 100% to 0%) to give pentyl 9-((2-hydroxyethyl)amino)nonanoate (3.05 g, 10.6 mmol, 65%) as a colorless oil. MS (ESI): m / z [M+H] + 288.2 for C 16 H 34 NO3; 1 H NMR (400 MHz, CDCl3)δ4.05 (t, J=6.7 Hz, 2H), 3.65-3.62 (m, 2H), 2.79-2.76 (m, 2H), 2.63-2.59 (m, 2H), 2.28 (t, J=7.5 Hz, 2H), 2.12 (bs, 2H), 1.65-1.58 (m, 4H), 1.50-1.45 (m, 2H), 1.34-1.30 (m, 12H), 0.90 (t, J=6.9 Hz, 3H); 13 C NMR(100 MHz, CDCl3)δ174.1, 64.5, 60.9, 51.1, 49.6, 34.2, 30.2, 29.5, 29.3, 29.2, 28.5, 28.2, 27.3, 25.1, 22.5, 14.1

[0180] Heptadecan-9-yl 5-bromopentanoate (Intermediate C) (Method C): [ka]

[0181] To crystals of 5-bromovaleric acid (10 g, 55.2 mmol, 1.00 equiv.) was added thionyl chloride (4.45 mL, 60.8 mmol, 1.1 equiv.) over 10 min with stirring at room temperature under an argon atmosphere. The mixture was heated to 80 °C and stirred for 1 h. Heptadecan-9-ol (14.2 g, 55.2 mmol, 1.0 equiv.) was then added to the reaction mixture over 1 h at 80 °C. The reaction mixture was then stirred at 100 °C for 4 h and at 50 °C for 16 h. The reaction mixture was then cooled to room temperature, diluted with EtOAc (100 mL), and subsequently washed with saturated sodium bicarbonate solution (3 × 50 mL), water (50 mL), and brine (50 mL). The organic layer was dried over sodium sulfate, and the solvent was evaporated. The residue (22.4 g) was purified on silica gel (hexane 100% to hexane:t-BuOMe, 90:10, v / v) to give heptadecan-9-yl 5-bromopentanoate (20.7 g, 48.9 mmol, 89%) as a pale yellow oil. 1 H NMR (600 MHz, CDCl3)δ4.89-4.85 (m, 1H), 3.41 (t, J=6.7 Hz, 2H), 2.33 (t, J=7.3 Hz, 2H), 1.93-1.88 (m, 2H), 1.81-1.77 (m, 2H), 1.51-1.49 (m, 4H), 1.30-1.26 (m, 24H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ173.1, 74.6, 34.3, 33.8, 33.2, 32.2, 32.0, 29.7, 29.6, 29.4, 25.5, 23.8, 22.8, 14.2.

[0182] Pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)amino)nonanoate (Intermediate D) (Method D): [ka]

[0183] To a mixture of pentyl 9-bromononanoate (Intermediate A) (0.50 g, 1.46 mmol, 1 equiv.) and tert-butyl (4-aminobutyl)carbamate (4.14 g, 22.0 mmol, 15 equiv.) was added a mixture of ethanol (5 mL) and acetonitrile (3 mL). The reaction mixture was stirred at 80 °C for 20 h. The reaction mixture was then cooled to room temperature, diluted with ethyl acetate (50 mL), and washed with water (2 × 100 mL) and brine (100 mL). The organic layer was dried over sodium sulfate and evaporated to dryness to give the crude product as a colorless oil (950 mg). The product was purified by silica gel chromatography (A phase: t-BuOMe; B phase t-BuOMe:MeOH:NH4OH = 50:50:1; 100% A to A:B = 50:50) to give pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)amino)nonanoate as a beige wax (580 mg, 1.40 mmol, 96%). MS (ESI): m / z [M+H] + 415.7 for C 23 H 47 N2O4; 1 H NMR (600 MHz, CDCl3) δ 4.92 (bs, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.12 (d, J=5.5 Hz, 2H, 2.61 (t, J=6.8 Hz, 2H), 2.57 (t, J=7.3 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H)p, 1.64-1.58 (m, 4H), 1.54-1.48 (m, 4H), 1.48-1.40 (m, 12H), 1.36-1.30 (m, 12H), 1.30-1.27 (m, 8H), 0.91 (t, J=7.1Hz, 3H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 156.0, 78.9, 64.4, 50.1, 49.6, 40.5, 34.4, 30.1, 29.4, 29.2, 29.1, 28.4, 28.3, 28.1, 27.9, 27.6, 27.3, 25.0, 22.3, 13.9.

[0184] Pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (Intermediate E) (Method E): [ka]

[0185] A mixture of heptadecan-9-yl 5-bromopentanoate (Intermediate C) (584 mg, 1.39 mmol, 1.05 equiv.) and pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)amino)nonanoate (Intermediate D) (550 mg, 1.33 mmol, 1.00 equiv.) was dissolved in a mixture of cyclopentyl methyl ether (2 mL) and acetonitrile (2 mL). Potassium carbonate (741 mg, 5.31 mmol, 4.00 equiv.) and potassium iodide (221 mg, 1.33 mmol, 1.00 equiv.) were then added to the mixture. The reaction mixture was stirred at 85 °C for 20 h. The reaction mixture was then cooled to room temperature, and the solvent was evaporated to dryness. The residue was dissolved in ethyl acetate (50 mL), and water (100 mL) was added. The aqueous layer was extracted with ethyl acetate (2 × 20 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, and evaporated to dryness to give the crude product as a pale yellow oil (1.28 g). The product was purified by silica gel chromatography (Phase A: t-BuOMe; Phase B t-BuOMe:MeOH:NH4OH = 50:50:1; 100% A to A:B = 50:50) to give pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (0.985 g, 1.31 mmol, 99%) as a colorless oil. MS (ESI): m / z [M+H] + 754.0 for C 46 H 89 N2O6; 1H NMR (600 MHz, CDCl3) δ 5.03 (s, 1H), 4.89-4.83 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.11 (bd, J=5.7 Hz, 2H), 2.39 (bs, 6H), 2.29 (t, J=7.4, 2H), 2.28 (t, J=7.6, 2H), 1.65-1.58 (m, 6H), 1.51-1.44 (m, 20H), 1.35-1.26 (m, 36 H), 0.90 (t, J=7.1 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.4, 156.0, 78.8, 74.2, 64.3, 54.0, 53.8, 53.6, 40.5, 34.6, 34.4, 34.1, 31.8, 29.5, 29.5, 29.4, 29.3, 29.2, 29.1, 28.4, 28.3, 28.1, 28.1, 27.5, 26.9, 26.4, 25.3, 25.0, 24.7, 23.2, 22.6, 22.3, 14.1, 13.9.

[0186] 3-Methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (Intermediate F): [ka]

[0187] To a suspension of 3,4-dimethoxy-3-cyclobutene-1,2-dione (10.0 g, 70.4 mmol) in diethyl ether (300 mL) was added a 2 M solution of methylamine in tetrahydrofuran (42 mL, 83.0 mmol, 1.2 equiv.) over 2 h. The reaction mixture was stirred at room temperature for 20 h. The volatiles were then evaporated under reduced pressure. The residue was washed with diethyl ether (3 × 20 mL), filtered off, and dried over sodium sulfate to give the product as a white solid (9.50 g, 67.3 mmol, 98%). Mp. 171-174 °C; 1H NMR (400 MHz, DMSO-d6) δ 8.21 (s, 1H), 4.29 (s, 3H), 3.39 (bs, 1H), 3.04 (d, J=4.7 Hz, 1H).

[0188] 5-Chloropentyl hexanoate (Intermediate G) (Method F): [ka]

[0189] A solution of 5-chloro-1-pentanol (3.00 g, 23.2 mmol, 1.0 equiv.), DIPEA (9.98 mL, 58.1 mmol, 2.5 equiv.), and 4-dimethylaminopyridine (143 mg, 1.16 mmol, 0.05 equiv.) in dichloromethane (50 mL) was cooled to 0 °C and flushed with argon. To the solution, hexanoyl chloride (3.32 mL, 23.2 mmol, 1 equiv.) was added over 10 min. The reaction mixture was then stirred at room temperature for 20 h. The reaction mixture was washed successively with 1 M hydrochloric acid solution (2 × 50 mL), water (50 mL), saturated sodium bisulfate solution (50 mL), and brine (50 mL). The organic layer was dried over sodium sulfate and evaporated to dryness. The product was purified by silica gel chromatography (100% hexane to 70:30 hexane:ethyl acetate) to give 5-chloropentyl hexanoate (3.85 g, 17.3 mmol, 99%) as a pale yellow oil. MS (APCI): m / z [M+H] + 220.8 for C 11 H 22 ClO2; 1 H NMR (600 MHz, CDCl3)δ4.08 (t, J=6.6 Hz, 2H), 3.54 (t, J=6.6 Hz, 2H), 2.30-2.28 (m, 2H), 1.83-1.78 (m, 2H), 1.67-1.61 (m, 4H), 1.53-1.50 (m, 2H), 1.35-1.28 (m, 4H), 0.89 (t, J=7.1 Hz, 3H); 13C NMR (150 MHz, CDCl3)δ174.1, 64.0, 44.9, 34.5, 32.3, 31.5, 28.1, 24.8, 23.5, 22.5, 14.0.

[0190] 9-Bromonoyl hexanoate (Intermediate H) (Method F) [ka]

[0191] A solution of 9-bromo-1-nonanol (3.00 g, 13.0 mmol, 1.00 equiv.), DIPEA (5.74 mL, 32.6 mmol, 2.50 equiv.), and 4-dimethylaminopyridine (80 mg, 0.65 mmol, 0.05 equiv.) in dichloromethane (50 mL) was cooled to 0 °C and flushed with argon. To the solution, hexanoyl chloride (1.79 g, 13.0 mmol, 1.00 equiv.) was added over 10 min. The reaction mixture was then stirred at room temperature over the weekend. The reaction mixture was subsequently washed with 1 M hydrochloric acid solution (2 × 50 mL), water (50 mL), saturated aqueous sodium bisulfate solution (50 mL), and brine (50 mL). The organic layer was dried over sodium sulfate and evaporated to dryness. The product was purified by silica gel chromatography (100% hexane to 70:30 hexane:ethyl acetate) to give 9-bromononyl hexanoate (2.52 g, 7.76 mmol, 59%) as a colorless oil. 1 H NMR (400 MHz, CDCl3)δ4.04 (t, J=6.7 Hz, 2H), 3.39 (t, J=6.9 Hz, 2H), 2.29-2.25 (m, 2H), 1.87-1.80 (m, 2H), 1.65-1.58 (m, 4H), 1.44-1.37 (m, 2H), 1.34-1.25 (m, 12H), 0.88 (t, J=7.0 Hz, 3H); 13 C NMR (100 MHz, CDCl3)δ174.1, 64.5, 34.5, 34.1, 32.9, 31.5, 29.4, 29.3, 28.8, 28.8, 28.3, 26.0, 24.9, 22.5, 14.1.

[0192] 11-Bromoundecylhexanoate (Intermediate I) (Method F) [ka]

[0193] A solution of 11-bromo-1-undecanol (3.00 g, 11.6 mmol, 1.00 equiv.), DIPEA (5.1 mL, 29.0 mmol, 2.50 equiv.), and 4-dimethylaminopyridine (71 mg, 0.58 mmol, 0.05 equiv.) in dichloromethane (50 mL) was cooled to 0 °C and flushed with argon. To the solution, hexanoyl chloride (1.59 g, 11.6 mmol, 1.00 equiv.) was added over 10 min. The reaction mixture was then stirred at room temperature over the weekend. The reaction mixture was subsequently washed with 1 M hydrochloric acid solution (2 × 50 mL), water (50 mL), saturated sodium bisulfate solution (50 mL), and brine (50 mL). The organic layer was dried over sodium sulfate and evaporated to dryness. The product was purified by silica gel chromatography (100% hexane to 70:30 hexane:ethyl acetate) to give 11-bromoundecyl hexanoate (2.52 g, 7.14 mmol, 61%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3)δ4.05 (t, J=6.7 Hz, 2H), 3.40 (t, J=6.9 Hz, 2H), 2.30-2.27 (m, 2H), 1.89-1.81 (m, 2H), 1.66-1.58 (m, 4H), 1.45-1.38 (m, 2H), 1.35-1.28 (m, 16H), 0.89 (t, J=7.0 Hz, 3H); 13 C NMR (100 MHz, CDCl3)δ174.2, 64.5, 34.5, 34.2, 33.0, 31.5, 29.6, 29.6, 29.5, 29.4, 28.9, 28.8, 28.3, 26.1, 24.9, 22.5, 14.1.

[0194] Pentyl 9-(3-hydroxypropylamino)nonanoate (Intermediate J) [ka]

[0195] Intermediate J was synthesized according to Representative Procedure 1 and General Methods A and B, starting with 3-hydroxy-1-aminopropane (7.00 equiv.) instead of ethanolamine. The product (pentyl 9-(4-hydroxybutylamino)nonanoate) (220 mg, 0.73 mmol, 83%) was obtained as a colorless oil. MS (ESI): m / z [M+H] + 302.4 for C 17 H 35 NO3; 1 H NMR (600 MHz, CDCl3)δ 4.04 (t, J=6.8 Hz, 2H), 3.79 (t, J=5.3 Hz, 2H), 3.00 (bs, 1H), 2.87-2.85 (zm, 2H), 2.58 (t, J=7.2 Hz, 2H), 2.27 (t, J=7.6 Hz, 2H), 1.67 (qu, J=5.5 Hz, 2H), 1.65-1.58 (m, 4H), 1.49-1.43 (m, 2H), 1.36-1.32 (m, 4H), 1.32-1.28 (m, 8H), 0.89 (t, J=7.1Hz, 3H); 13 C NMR(151 MHz, CDCl3)δ173.9, 64.5, 64.3, 50.1, 49.8, 34.3, 30.6, 29.8, 29.3, 29.1, 29.0, 28.3, 28.0, 27.1, 24.9, 22.3, 13.9.

[0196] Pentyl 9-(4-hydroxybutylamino)nonanoate (Intermediate K) [ka]

[0197] Intermediate K was synthesized according to Representative Procedure 1 and General Methods A and B, starting with 4-hydroxy-1-aminobutane (5.00 equiv.) instead of ethanolamine. The product (pentyl 9-(4-hydroxybutylamino)nonanoate) (150 mg, 0.48 mmol, 81%) was obtained as a colorless oil. MS (ESI): m / z [M+H] + 316.4 for C 18 H 37 NO3; 1 H NMR (600 MHz, CDCl3)δ 4.04 (t, J=6.8 Hz, 2H), 3.55 (t, J=5.2 Hz, 2H), 2.62 (t, J=5.5 Hz, 2H), 2.57 (t, J=7.3 Hz, 2H), 2.26 (t, J=7.5 Hz, 2H), 1.70-1.65 (m, 2H), 1.65-1.58 (m, 4H), 1.52-1.45 (m, 2H), 1.37-1.31 (m, 4H), 1.3-1.27 (m, 8H), 0.89 (t, J=7.1 Hz, 3H); 13 C NMR(151 MHz, CDCl3)δ173.9, 64.3, 62.5, 49.6, 49.5, 34.3, 32.7, 29.7, 29.3, 29.1, 29.0, 28.9, 28.3, 28.0, 27.2, 24.9, 22.3, 13.9.

[0198] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2 hydroxyethyl)amino)nonanoate (Compound (I)(a)) (Compound 1) (Method G): [ka]

[0199] To a mixture of pentyl 9-((2-hydroxyethyl)amino)nonanoate (Intermediate B) (350 mg, 1.22 mmol, 1.00 equiv.) and heptadecan-9-yl 5-bromopentanoate (Intermediate C) (536 mg, 1.28 mmol, 1.05 equiv.), acetonitrile (10 mL) and cyclopentyl methyl ether (15 mL) were added. Next, potassium carbonate (680 mg, 4.87 mmol, 4.00 equiv.) and potassium iodide (223 mg, 1.34 mmol, 1.10 equiv.) were added. The reaction mixture was stirred at 80 °C for 4 h and then at 60 °C for 2 days. The reaction mixture was cooled, and the solid was filtered and washed with acetonitrile (25 mL). The filtrate was evaporated and the residue was purified on silica gel (A phase: DCM 100%, B phase: DCM / MeOH / NH4OH, 80:20:1, v / v / v; A:B, 100% to 0%) to give pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (490 mg, 0.78 mmol, 64%) as a colorless oil. MS (ESI): m / z [M+H] + 626.6 for C 38 H 76 NO5; 1 H NMR (400 MHz, CDCl3)δ4.86 (qu, J=6.3 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.56 (t, J=5.1 Hz, 2H), 2.62 (t, J=4.9 Hz, 2H), 2.50 (dt, J=14.9, 7.5 Hz, 4H), 2.29 (q, J=7.5 Hz, 4H), 1.65-1.58 (m, 6H), 1.55-1.46 (m, 8H), 1.34-1.25 (m, 36H), 0.92-0.86 (m, 9H); 13C NMR (100 MHz, CDCl3)δ174.1, 173.4, 74.5, 64.5, 58.4, 55.9, 54.0, 53.7, 34.5, 34.3, 32.0, 29.7, 26.6, 29.5, 29.4, 29.3, 28.5, 28.2, 27.5, 26.9, 26.4, 25.5, 25.1, 23.1, 22.8, 22.5, 14.2, 14.1.

[0200] Pentyl 6-{[5-(heptadecan-9-yloxy)-5-oxopentyl](2-hydroxyethyl)amino}hexanoate (compound 2): [ka]

[0201] Compound 2 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (491 mg, 0.776 mmol, 65%). MS (ESI): m / z [M+H] + 584.9 for C 35 H 70 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.53 (t, J=5.3 Hz, 2H), 2.57 (t, J=5.3 Hz, 2H), 2.49-2.44 (m, 4H), 2.31-2.28 (m, 4H), 1.65-1.58 (m, 6H), 1.50-1.43 (m, 8H), 1.35-1.25 (m, 31H), 0.90 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H). 13C NMR (150 MHz, CDCl3) δ 173.9, 173.4, 74.4, 64.6, 58.5, 55.7, 53.7, 53.6, 34.6, 34.4, 34.3, 32.0, 29.7, 29.6, 29.4, 28.5, 28.2, 27.1, 27.0, 26.7, 25.5, 25.0, 23.1, 22.8, 22.5, 14.2, 14.1.

[0202] Pentyl 7-{[5-(heptadecan-9-yloxy)-5-oxopentyl](2-hydroxyethyl)amino}heptanoate (compound 3): [ka]

[0203] Compound 3 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (246 mg, 0.411 mmol, 71%). MS (ESI): m / z [M+H] + 598.5 for C 36 H 72 NO5; 1 H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (p, J=6.2 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.62 (t, J=5.8 Hz, 2H), 2.67 (t, J=5.8 Hz, 2H), 2.63-2.49 (m, 4H), 2.33 (dt, J=9.7, 7.4 Hz, 4H), 1.70-1.59 (m, 6H), 1.58-1.44 (m, 8H), 1.41-1.21 (m, 32H), 0.92 (t, J=7.0 Hz, 3H), 0.82 (t, J=6.9 Hz, 3H); 13C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 174.9, 174.3, 75.1, 65.0, 58.9, 56.0, 54.4, 54.1, 34.7, 34.6, 34.4, 32.2, 29.8, 29.7, 29.6, 29.3, 28.6, 28.4, 27.4, 26.5, 26.1, 25.7, 25.2, 23.3, 23.0, 22.6, 14.2, 14.1.

[0204] Pentyl 8-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)octanoate (compound 4): [ka]

[0205] Compound 4 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (230 mg, 0.372 mmol, 68%). MS (ESI): m / z [M+H] + 613.1 for C 37 H 74 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.52 (t, J=5.4 Hz, 2H), 2.57 (t, J=5.4 Hz, 2H), 2.49-2.42 (m, 4H), 2.31-2.27 (m, 4H), 1.64-1.59 (m, 6H), 1.51-1.46 (m, 6H), 1.45-1.40 (m, 2H), 1.36-1.25 (m, 35H), 0.90 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.1 Hz, 6H); 13C NMR (150 MHz, CDCl3) δ 174.1, 173.5, 74.4, 64.6, 58.5, 55.7, 53.9, 53.6, 34.6, 34.5, 34.3, 32.0, 29.7, 29.7, 29.4, 29.4, 29.3, 28.5, 28.2, 27.4, 27.2, 26.8, 25.5, 25.1, 23.1, 22.8, 22.5, 14.2, 14.1.

[0206] Pentyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)decanoate (compound 5): [ka]

[0207] Compound 5 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (330 mg, 0.516 mmol, 78%). MS (ESI): m / z [M+H] + 640.9 for C 39 H 78 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.87 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.52 (t, J=5.4 Hz, 2H), 2.90 (s, 1H), 2.57 (t, J=5.4 Hz, 2H), 2.47 (t, J=7.4 Hz, 2H), 2.43 (t, J=7.5 Hz, 2H), 2.31 (t, J=7.7 Hz, 2H), 2.30 (t, J=7.6 Hz, 2H), 1.65-1.59 (m, 6H), 1.51-1.46 (m, 6H), 1.45-1.40 (m, 2H), 1.38-1.34 (m, 6H), 1.30-1.26 (m, 32H), 0.91 (t, J=7.0 Hz, 3H), 0.89 (t, J=7.1 Hz, 6H); 13C NMR (150 MHz, CDCl3) δ 173.9, 173.3, 74.2, 64.3, 58.4, 55.5, 53.8, 53.4, 34.4, 34.3, 34.1(2x), 31.8(2x), 29.5(3x), 29.5(2x), 29.4, 29.2(3x), 29.1, 28.3, 28.1, 27.4, 27.2, 26.7, 25.3(2x), 25.0, 23.0, 22.6(2x), 22.3, 14.1(2x), 13.9.

[0208] Pentyl 11-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)undecanoate (compound 6): [ka]

[0209] Compound 6 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (337 mg, 0.52 mmol, 81%). MS (ESI): m / z [M+H] + 655.1 for C 40 H 80 NO5; 1 H NMR (600 MHz, CDCl3)δ4.86 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.58 (t, J=6.1 Hz, 2H), 2.65 (t, J=5.1 Hz, 2H), 2.60-2.48 (m, 4H), 2.29 (dt, J=14.9, 7.4 Hz, 4H), 1.66-1.57 (m, 6H), 1.57-1.43 (m, 8H), 1.36-1.31 (m, 4H), 1.31-1.20 (m, 36H), 0.90 (t, J=7.0 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13C NMR (150 MHz, CDCl3)δ174.1, 173.2, 74.6, 64.5, 58.3, 55.9, 53.9, 53.2, 34.5, 34.2, 32.2, 31.9, 29.6, 29.6, 29.6, 29.5, 29.3, 29.2, 28.4, 28.2, 27.4, 25.4, 25.1, 22.7, 22.4, 14.2, 14.0.

[0210] Pentyl 12-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)dodecanoate (compound 7): [ka]

[0211] Compound 7 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (270 mg, 0.404 mmol, 67%). MS (ESI): m / z [M+H] + 668.9 for C 41 H 82 NO5; 1 H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (p, J=6.2 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.61 (t, J=5.8 Hz, 2H), 2.65 (t, J=5.7 Hz, 2H), 2.60-2.48 (m, 4H), 2.33 (dt, J=13.2, 7.3 Hz, 4H), 1.70-1.58 (m, 6H), 1.57-1.42 (m, 8H), 1.39-1.21 (m, 42H), 0.92 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 174.5, 173.7, 74.5, 64.4, 58.4, 55.4, 53.9, 53.5, 34.2, 34.1, 33.8, 31.6, 29.3, 29.3, 29.3, 29.2, 29.2, 29.0, 28.8, 28.0, 27.8, 27.2, 26.2, 25.6, 25.1, 24.7, 22.7, 22.4, 22.0, 13.6, 13.5

[0212] Pentyl 13-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)tridecanoate (compound 8): [ka]

[0213] Compound 8 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (160 mg, 0.235 mmol, 54%). MS (ESI): m / z [M+H] + 682.8 for C 42 H 84 NO5; 1 H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (qu, J=6.2 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.61 (t, J=5.8 Hz, 2H), 2.64 (t, J=5.9 Hz, 2H), 2.58-2.46 (m, 4H), 2.33 (dt, J=12.5, 7.4 Hz, 4H), 1.69-1.58 (m, 6H), 1.58-1.39 (m, 10H), 1.39-1.17 (m, 42H), 0.91 (t, J=7.0 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H).

[0214] Pentyl 14-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)tetradecanoate (compound 9): [ka]

[0215] Compound 9 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (264 mg, 0.380 mmol, 68%). MS (ESI): m / z [M+H] + 696.9 for C 43 H 86 NO5; 1 H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (qu, J=6.2 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.61 (t, J=5.9 Hz, 2H), 2.64 (t, J=5.9 Hz, 2H), 2.58-2.47 (m, 4H), 2.33 (dt, J=12.7, 7.3 Hz, 4H), 1.71-1.58 (m, 6H), 1.58-1.41 (m, 10H), 1.39-1.21 (m, 44H), 0.92 (t, J=7.0 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 175.2, 174.3, 75.1, 65.0, 59.1, 56.0, 54.5, 54.1, 34.8, 34.7, 34.4, 32.2, 30.0, 29.9, 29.9, 29.8, 29.8, 29.7, 29.6, 29.6, 29.6, 29.4, 28.6, 28.4, 27.8, 26.8, 26.3, 25.7, 25.3, 23.3, 23.0, 22.6, 14.2, 14.1

[0216] Pentyl 15-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)pentadecanoate (compound 10): [ka]

[0217] Compound 10 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (315 mg, 0.444 mmol, 83%). MS (ESI): m / z [M+H] + 710.7 for C 44 H 88 NO5; 1 H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.88 (p, J=6.3 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.65 (t, J=5.8 Hz, 2H), 2.77-2.69 (m, 2H), 2.68-2.56 (m, 4H), 2.39-2.28 (m, 4H), 1.69-1.59 (m, 6H), 1.59-1.46 (m, 8H), 1.38-1.23 (m, 48H), 0.95-0.86 (m, 9H).

[0218] Pentyl 9-((2-(heptadecan-9-yloxy)-2-oxoethyl)(2-hydroxyethyl)amino)nonanoate (compound 11): [ka]

[0219] Compound 11 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (225 mg, 0.385 mmol, 73%). MS (ESI): m / z [M+H] + 585.0 for C 35 H 70 NO5; 1H NMR (400 MHz, CDCl3) δ 4.92 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.57 (t, J=5.0 Hz, 2H), 3.38 (s, 2H), 2.81 (t, J=4.5 Hz, 2H), 2.66 (t, J=7.3 Hz, 2H), 2.28 (t, J=7.5 Hz, 2H), 1.66-1.57 (m, 4H), 1.53-1.47 (m, 6H), 1.35-1.26 (m, 36H), 0.92-0.86 (m, 9H); 13 C NMR (100 MHz, CDCl3) δ 173.9, 171.5, 75.2, 64.4, 58.8, 56.9, 55.0, 54.6, 34.4, 34.1 (2x), 31.8 (2x), 29.5 (2x), 29.3, 29.2, 29.2 (2x), 28.3, 28.1, 27.6, 27.1, 25.3 (2x), 25.0, 22.6 (2x), 22.3, 14.1 (2x), 13.9.

[0220] Pentyl 9-((3-(heptadecan-9-yloxy)-3-oxopropyl)(2-hydroxyethyl)amino)nonanoate (compound 12): [ka]

[0221] Compound 12 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as (mg, mmol, %). MS (ESI): m / z [M+H] + 598.6 for C 36 H 72 NO5; 1H NMR (600 MHz, CDCl3) δ 4.90-4.86 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.55 (t, J=5.2 Hz, 2H), 2.80 (t, J=6.9 Hz, 2H), 2.59-2.57 (m, 2H), 2.45-2.42 (m, 4H), 2.30-2.27 (m, 2H), 1.65-1.59 (m, 4H), 1.52-1.51 (m, 4H), 1.45-1.32 (m, 6H), 1.29-1.26 (m, 36H), 0.91 (t, J=7.1 Hz, 3H), 0.88 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.1, 172.7, 74.8, 64.5, 59.0, 55.8, 54.1, 49.5, 34.5, 34.2, 33.1, 32.0 ,29.7, 29.6, 29.5, 29.4, 29.4, 29.3, 28.5, 28.2, 27.5, 27.3, 25.5, 25.1, 22.8, 22.5, 14.2, 14.1.

[0222] Pentyl 9-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)nonanoate (compound 13): [ka]

[0223] Compound 13 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (220 mg, 0.356 mmol, 47%). MS (ESI): m / z [M+H] + 612.5 for C 37 H 74 NO5; 1H NMR (400 MHz, CDCl3) δ 4.86 (qu, J=6.3 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.57 (t, J=5.2 Hz, 2H), 2.63 (t, J=4.9 Hz, 2H), 2.56-2.48 (m, 4H), 2.30 (dt, J=10.0, 7.4 Hz, 4H), 1.84-1.76 (m, 2H), 1.64-1.58 (m, 4H), 1.51-1.46 (m, 6H), 1.34-1.25 (m, 36H), 0.92-0.86 (m, 9H). 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.3, 74.7, 64.5, 58.5, 56.0, 54.0, 53.2, 34.5, 34.2, 32.3, 32.0, 29.7, 29.7, 29.5, 29.4, 29.3, 28.5, 28.2, 27.5, 26.9, 25.5, 25.1, 22.8, 22.5, 14.2, 14.1.

[0224] Pentyl 10-((2-(heptadecan-9-yloxy)-2-oxoethyl)(2-hydroxyethyl)amino)decanoate (compound 14): [ka]

[0225] Compound 14 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (290 mg, 0.485 mmol, 73%). MS (ESI): m / z [M+H] + 598,9 for C 36 H 72 NO5; 1H NMR (400 MHz, CDCl3) δ 4.92 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.56 (bt, J=5.2 Hz, 2H), 3.36 (s, 2H), 3.22 (bs, 1H), 2.80 (t, J=5.2 Hz, 2H), 2.64 (t, J=7.5 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.62-1.62 (m, 4H), 1.53-1.52 (m, 4H), 1.45-1.45 (m, 2H), 1.35-1.25 (m, 38H), 0.91 (t, J=6.9 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 173.9, 171.7, 75.0, 64.3, 58.9, 56.9, 55.1, 54.6, 34.4, 34.0, 31.8 (2x), 29.5 (3x), 29.4, 29.2(2x), 29.2 (2x), 29.1, 28.3, 28.1, 27.7, 27.1, 25.3 (2x), 25.0, 22.6 (2x), 22.3, 14.1 (2x), 13.9.

[0226] Pentyl 10-((3-(heptadecan-9-yloxy)-3-oxopropyl)(2-hydroxyethyl)amino)decanoate (compound 15): [ka]

[0227] Compound 15 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (150 mg, 0.245 mmol, 36%). MS (ESI): m / z [M+H] + 612.9 for C 37 H 74 NO5; 1H NMR(600 MHz, CDCl3) δ 4.88 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.58 (bs, 2H), 2.84 (bs, 2H), 2.62 (bs, 2H), 2.47 (bs, 4H), 2.29 (t, J=7.8 Hz, 2H), 1.57-1.68 (m, 4H), 1.47-1.57 (m, 4H), 1.39-1.47 (bs, 2H), 1.32-1.39 (m, 6H), 1.13-1.32 (m, 34H), 0.91 (t, J=7.1 Hz, 3H), 0.88 (t, J=7.1 Hz, 6H); 13 C NMR(150 MHz, CDCl3) δ 173.9, 172.4, 74.8, 64.4, 58.7, 55.8, 54.0, 49.4, 34.4, 34.0 (2x), 32.7, 31.8 (2x), 29.5 (2x), 29.5 (2x), 29.4, 29.2 (3x), 29.1, 28.3, 28.1, 27.3, 26.9, 25.3(2x), 25.0, 22.6 (2x), 22.3, 14.1 (2x), 13.9.

[0228] Pentyl 6-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)hexanoate (compound 16): [ka]

[0229] Compound 16 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (110 mg, 0.191 mmol, 31%). MS (ESI): m / z [M+H] + 570.9 for C 34 H 68 NO5; 1H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.61 (bs, 2H), 2.68 (bs, 2H), 2.60-2.56 (m, 3H), 2.32 (t, J=7.2 Hz, 2H), 2.30 (t, J=7.5 Hz, 2H), 1.83 (bs, 2H), 1.66-1.60 (m, 4H), 1.54-1.50 (m, 6H), 1.36-1.25 (m, 32H), 0.90 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.8, 173.2, 74.8, 64.6, 58.4, 56.2, 53.9, 53.3, 34.3, 34.2, 32.2, 32.0, 29.7, 29.4, 28.5, 28.2, 27.0, 25.5, 24.9, 22.8, 22.5, 14.2, 14.1.

[0230] Pentyl 8-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)octanoate (compound 17): [ka]

[0231] Compound 17 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (200 mg, 0.331 mmol, 60%). MS (ESI): m / z [M+H] + 598.9 for C 36 H 72 NO5; 1H NMR (600 MHz, CDCl3) δ 4.87-4.83 (m, 1H), 4.04 (t, J=6.8 Hz, 2H), 3.61 (bs, 2H), 2.69-2.56 (m, 5H), 2.31 (t, J=7.1 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H), 1.83 (bs, 2H), 1.63-1.57 (m, 4H), 1.50-1.49 (m, 6H), 1.34-1.24 (m, 36H), 0.89 (t, J=7.0 Hz, 3H), 0.86 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.0, 173.2, 74.8, 64.6, 58.3, 56.3, 54.1, 53.3, 34.5, 34.2, 32.0, 29.7, 29.4, 29.2, 28.5, 28.2, 27.3, 25.5, 25.1, 22.8, 22.5, 14.2, 14.1.

[0232] Pentyl 10-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)decanoate (compound 18): [ka]

[0233] Compound 18 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (330 mg, 0.527 mmol, 79%). MS (ESI): m / z [M+H] + 627.1 for C 38 H 76 NO5; 1H NMR (600 MHz, CDCl3) δ 4.87 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.53 (t, J=5.4 Hz, 2H), 2.59 (t, J=5.4 Hz, 2H), 2.49 (t, J=7.3 Hz, 2H), 2.45 (t, J=7.5 Hz, 2H), 2.30 (t, J=7.6 Hz, 2H), 2.29 (t, J=7.7, 2H), 1.77 (qu, J=7.3 Hz, 2H), 1.65-1.59 (m, 4H), 1.51-1.50 (m, 4H), 1.42 (qu, J=7.2 Hz, 2H), 1.38-1.32 (m, 4H), 1.30-1.26 (m, 34H),0.91 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.3, 74.4, 64.3, 58.5, 55.6, 53.7, 53.0, 34.4, 34.1 (2x), 32.3, 31.8(2x), 29.5 (3x), 29.5(2x), 29.4, 29.2(3x), 29.1, 28.3, 28.1, 27.4, 27.1, 25.3(2x), 25.0, 22.6(2x), 22.6, 22.3, 14.1(2x), 13.9.

[0234] Pentyl 11-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)undecanoate (compound 19): [ka]

[0235] Compound 19 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (323 mg, 0.51 mmol, 80%). MS (ESI): m / z [M+H] + 641.1 for C 39 H 77 NO5;1 H NMR (600 MHz, CDCl3)δ4.86 (p, J=6.3 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.58 (t, J=4.5 Hz, 2H), 2.65 (t, J=5.1 Hz, 2H), 2.59-2.54 (m, 2H), 2.53-2.48 (m, 2H), 2.30 (dt, J=18.4, 7.4 Hz, 4H), 1.81 (p, J=7.2 Hz, 2H), 1.65-1.57 (m, 4H), 1.54-1.43 (m, 6H), 1.36-1.30 (m, 4H), 1.30-1.20 (m, 36H), 0.90 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.1, 173.2, 74.6, 64.5, 58.3, 55.9, 53.9, 53.2, 34.5, 34.2, 32.2, 31.9, 29.6, 29.6, 29.6, 29.5, 29.3, 29.2, 28.4, 28.2, 27.4, 25.4, 25.1, 22.7, 22.4, 14.2, 14.0

[0236] Pentyl 12-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)dodecanoate (compound 20): [ka]

[0237] Compound 20 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (264 mg, 0.404 mmol, 67%). MS (ESI): m / z [M+H] + for C 40 H 80 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.88 (p, J=6.2 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.62 (t, J=5.7 Hz, 2H), 2.68 (t, J=5.3 Hz, 2H), 2.62-2.51 (m, 4H), 2.38-2.29 (m, 4H), 1.86-1.78 (m, 2H), 1.68-1.59 (m, 4H), 1.58-1.51 (m, 4H), 1.50-1.46 (m, 2H), 1.39-1.22 (m, 42H), 0.92 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 174.5, 173.5, 74.7, 64.4, 58.5, 55.5, 54.0, 53.1, 34.2, 33.8, 32.0, 31.6, 29.4, 29.3, 29.3, 29.2, 29.2, 29.2, 29.0, 28.9, 28.0, 27.8, 27.2, 26.2, 25.1, 24.8, 22.4, 22.0, 21.7, 13.6, 13.5.

[0238] Pentyl 15-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)pentadecanoate (compound 21): [ka]

[0239] Compound 21 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (270 mg, 0.388 mmol, 72%). MS (ESI): m / z [M+H] + for C 43 H 85 NO5; 1H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.88 (qu, J=6.3 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.61 (t, J=5.9 Hz, 2H), 2.69-2.63 (m, 2H), 2.60-2.49 (m, 4H), 2.38-2.27 (m, 4H), 1.87-1.76 (m, 2H), 1.68-1.58 (m, 4H), 1.57-1.43 (m, 6H), 1.38-1.23 (m, 48H ), 0.95-0.86 (m, 9H); 13 C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 174.5, 173.6, 74.6, 64.3, 58.5, 55.5, 54.0, 53.1, 34.1, 33.8, 32.0, 31.6, 29.4, 29.3, 29.3, 29.2, 29.2, 29.0, 28.8, 28.0, 27.8, 27.2, 26.2, 25.1, 24.7, 22.3, 22.0, 21.7, 13.6, 13.5.

[0240] Methyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 22): [ka]

[0241] Compound 22 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (441 mg, 0.77 mmol, 90%). MS (ESI): m / z [M+H] + 571.0 for C 34 H 68 NO5; 1H NMR (400 MHz, CDCl3)δ4.86 (qu, J=6.3 Hz, 1H), 3.66 (s, 3H), 3.53 (t, J=5.4 Hz, 2H), 2.58 (t, J=5.4 Hz, 2H), 2.52-2.41 (m, 4H), 2.32-2.26 (m, 4H), 1.66-1.56 (m, 4H), 1.54-1.38 (m, 8H), 1.32-1.19 (m, 32H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3)δ174.4, 173.4, 74.4, 58.4, 55.6, 53.9, 53.5, 51.5, 34.5, 34.2, 34.2, 31.9, 29.6, 29.6, 29.4, 29.3, 29.3, 29.2, 27.4, 27.1, 26.6, 25.4, 25.0, 23.0, 22.7, 14.2.

[0242] Ethyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 23): [ka]

[0243] Compound 23 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (408 mg, 0.70 mmol, 91%). MS (ESI): m / z [M+H] + 585.0 for C 35 H 70 NO5; 1H NMR (600 MHz, CDCl3)δ4.86 (qu, J=6.3 Hz, 1H), 4.12 (q, J=7.1 Hz, 2H), 3.59 (t, J=4.4 Hz, 2H), 2.67-2.63 (m, 2H), 2.58-2.49 (m, 4H), 2.29 (dt, J=17.3, 7.4 Hz, 4H), 1.65-1.57 (m, 4H), 1.56-1.44 (m, 8H), 1.32-1.20 (m, 35H), 0.87 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ173.9, 173.3, 74.4, 60.2, 58.2, 55.9, 54.0, 53.6, 34.4, 34.2, 31.9, 29.6, 29.6, 29.4, 29.3, 29.3, 29.1, 27.3, 25.4, 25.0, 22.9, 22.7, 14.3, 14.2.

[0244] Propyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 24): [ka]

[0245] Compound 24 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (401 mg, 0.67 mmol, 87%). MS (ESI): m / z [M+H] + 599.0 for C 36 H 72 NO5; 1H NMR (600 MHz, CDCl3)δ4.86 (p, J=6.2 Hz, 1H), 4.02 (t, J=6.7 Hz, 2H), 3.58 (t, J=5.1 Hz, 2H), 2.65 (t, J=4.5 Hz, 2H), 2.58-2.48 (m, 4H), 2.30 (dt, J=10.7, 7.4 Hz, 4H), 1.67-1.58 (m, 6H), 1.56-1.44 (m, 8H), 1.32-1.21 (m, 32H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.0, 173.3, 74.4, 66.9, 58.2, 55.9, 54.0, 53.6, 34.4, 34.2, 31.9, 29.6, 29.6, 29.4, 29.3, 29.3, 29.2, 27.4, 25.4, 25.0, 22.9, 22.7, 22.0, 14.2, 10.5.

[0246] Butyl 9-((2-(heptadecan-9-yloxy)-2-oxoethyl)(2-hydroxyethyl)amino)nonanoate (compound 25): [ka]

[0247] Compound 25 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (200 mg, 0.351 mmol, 41%). MS (ESI): m / z [M+H] + 570.4 for C 34 H 68 NO5; 1H NMR (400 MHz, CDCl3) δ 4.95-4.89 (m, 1H), 4.07 (t, J = 6.7 Hz, 2H), 3.56-3.53 (m, 2H), 3.34 (s, 2H), 2.79-2.76 (m, 2H), 2.64-2.60 (m, 2H), 2.30-2.27 (m, 2H), 1.64-1.57 (m, 4H), 1.53-1.52 (m, 4H), 1.46-1.35 (m, 4H), 1.29-1.26 (m, 35H), 0.93 (t, J=7.4 Hz, 3H), 0.88 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 172.0, 75.2, 64.2, 59.1, 57.0, 55.4, 54.7, 34.5, 34.2, 32.0, 30.8, 29.6, 29.5, 29.4, 29.4, 29.3, 27.9, 27.3, 25.5, 25.1, 22.8, 19.3, 14.2, 13.8.

[0248] Butyl 9-((3-(heptadecan-9-yloxy)-3-oxopropyl)(2-hydroxyethyl)amino)nonanoate (compound 26): [ka]

[0249] Compound 26 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (100 mg, 0.171 mmol, 34%). MS (ESI): m / z [M+H] + 584.9 for C 35 H 70NO5; 1H-NMR (400 MHz, CDCl3) δ 4.88 (qu, J=6.3 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.58 (t, J=4.5 Hz, 2H), 2.82 (t, J=6.1 Hz, 2H), 2.61 (bs, 2H), 2.46 (bs, 4H), 2.29 (t, J=7.6 Hz, 2H), 1.64-1.57 (m, 4H), 1.52-1.50 (m, 5H), 1.41-1.33 (m, 3H), 1.30-1.26 (m, 32H), 0.93 (t, J=7.4 Hz, 3H), 0.88 (t, J=6.8 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 173.9, 172.5, 74.8, 64.1, 58.7, 55.8, 54.0, 49.4, 34.4, 34.0 (2x), 31.8 (2x), 30.7, 29.5 (2x), 29.5 (2x), 29.4, 29.2 (3x), 29.1, 27.3, 25.3 (2x), 25.0, 22.6 (2x), 19.1, 14.1 (2x), 13.7.

[0250] Butyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 27): [ka]

[0251] Compound 27 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (172 mg, 0.281 mmol, 59%). MS (ESI): m / z [M+H] + 613.1 for C 37 H 74 NO5; 1H NMR (400 MHz, CDCl3) δ 4.90-4.84 (m, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.52 (t, J=5.4 Hz, 2H), 2.57 (t, J=5.4 Hz, 2H), 2.49-2.41 (m, 4H), 2.32-2.27 (m, 4H), 1.66-1.57 (m, 6H), 1.51-1.35 (m, 10H), 1.29-1.26 (m, 35H), 0.93 (t, J=7.4 Hz, 3H), 0.88 (t, J=6.9 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.1, 173.5, 74.4, 64.2, 58.5, 55.6, 53.9, 53.6, 34.6, 34.5, 34.3, 32.0, 30.8, 29.7, 29.6, 29.4, 29.4, 29.3, 27.5, 27.3, 26.8, 25.5, 25.1, 23.1, 22.8, 19.3, 14.2, 13.8.

[0252] Methyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)decanoate (compound 28): [ka]

[0253] Compound 28 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (140 mg, 0.237 mmol, 39%). MS (ESI): m / z [M+H] + 584.9 for C 35 H 70 NO5; 1H NMR (400 MHz, CDCl3) δ 4.86 (p, J=6.3 Hz, 1H), 3.66 (bs, 5H), 2.47-2.63 (m, 5H), 2.34-2.28 (m, 4H), 1.63-1.40 (m, 12H), 1.28-1.25 (m, 35H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 174.4, 173.2, 74.6, 58.0, 54.2, 53.8, 51.6, 34.3, 34.2, 32.0, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2, 27.3, 25.5, 25.1, 22.8, 14.2.

[0254] Methyl 10-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)decanoate (compound 29): [ka]

[0255] Compound 29 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (140 mg, 0.243 mmol, 40%). MS (ESI): m / z [M+H] + 570.7 for C 34 H 68 NO5; 1 H NMR (400 MHz, CDCl3)δ4.86 (qu,J=6.3 Hz, 1H), 3.66 (s, 3H), 3.64 (bs, 2H), 2.72-2.59 (m, 5H), 2.35-2.28 (m, 4H), 1.87-1.84 (m, 2H), 1.63-1.57 (m, 2H), 1.51-1.40 (m, 6H), 1.28-1.25 (m, 35H), 0.87 (t,J=6.9 Hz, 6H); 13C NMR (100 MHz, CDCl3) δ 174.4, 173.1, 74.8, 58.2, 56.4, 54.1, 53.3, 51.6, 34.2, 32.1, 32.0, 29.7, 29.6, 29.5, 29.4, 29.3, 29.2, 27.4, 25.5, 25.1, 22.8, 14.8.

[0256] Ethyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)decanoate (compound 30): [ka]

[0257] Compound 30 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (200 mg, 0.331 mmol, 57%). MS (ESI): m / z [M+H] + 598.3 for C 36 H 72 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.12 (q, J=7.1 Hz, 2H), 3.68 (bs, 2H), 2.76-2.66 (m, 4H), 2.33-2.31 (m, 2H), 2.29-2.27 (m, 2H), 1.63-1.50 (m, 11H), 1.31-1.24 (m, 38H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.0, 173.2, 74.6, 60.3, 57.9, 56.5, 54.2, 53.8, 34.5, 34.3, 32.0, 29.7, 29.6, 29.5, 29.5, 29.4, 29.3, 29.2, 25.5, 25.1, 22.8, 14.4, 14.2.

[0258] Ethyl 10-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)decanoate (compound 31): [ka]

[0259] Compound 31 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (190 mg, 0.322 mmol, 56%). MS (ESI): m / z [M+H] + 584.9 for C 35 H 70 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.12 (q, J=7.1 Hz, 2H), 3.68-3.62 (m, 2H), 2.75-2.62 (m, 4H), 2.34 (t, J=6.8 Hz, 2H), 2.29-2.27 (m, 2H), 1.89 (bs, 2H), 1.63-1.59 (m 2H), 1.52-1.50 (m, 5H), 1.31-1.24 (m, 38H), 0.87 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.0, 173.0, 74.9, 60.3, 58.1, 54.2, 53.3, 34.5, 34.2, 32.0, 29.7, 29.5, 29.4, 29.3, 29.2, 25.5, 25.1, 22.8, 14.4, 14.2.

[0260] Propyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)decanoate (compound 32): [ka]

[0261] Compound 32 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (170 mg, 0.275 mmol, 50%). MS (ESI): m / z [M+H] + 612.3 for C 37 H 74 NO5; 1 H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.02 (t, J=6.7 Hz, 2H), 3.66-3.64 (m, 2H), 2.73-2.60 (m, 5H), 2.32 (t, J=7.3 Hz, 2H), 2.30-2.28 (m, 2H), 1.67-1.59 (m, 7H), 1.52-1.50 (m, 6H), 1.30-1.25 (m, 37H), 0.94 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.0, 173.2, 74.5, 65.9, 58.0, 56.2, 54.1, 53.7, 34.4, 34.3, 34.2, 31.9, 29.6, 29.6, 29.4, 29.3, 29.3, 29.2, 27.3, 25.4, 25.1, 22.8, 22.7, 22.1, 14.2, 10.5.

[0262] Propyl 10-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)decanoate (compound 33): [ka]

[0263] Compound 33 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (170 mg, 0.281 mmol, 51%). MS (ESI): m / z [M+H] + 598.3 for C 36 H 72 NO5; 1H NMR (600 MHz, CDCl3) δ 4.88-4.84 (m, 1H), 4.02 (t, J=6.7 Hz, 2H), 3.62 (bs, 2H), 2.72-2.57 (m, 5H), 2.32 (t, J=7.1 Hz, 2H), 2.30-2.28 (m, 2H), 1.84 (bs, 2H), 1.67-1.59 (m, 4H), 1.51-1.50 (m, 6H), 1.30-1.25 (m, 37H), 0.94 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.1, 173.2, 74.8, 66.0, 58.3, 56.3, 54.1, 53.3, 34.5, 34.2, 32.1, 32.0, 29.7, 29.6, 29.5, 29.4, 29.3, 29.3, 27.4, 25.5, 25.1, 22.8, 22.2, 14.2, 10.5.

[0264] Butyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)decanoate (compound 34): [ka]

[0265] Compound 34 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (450 mg, 0.712 mmol, 68.2%). MS (ESI): m / z [M+H] + 626.6 for C 38 H 76 NO5; 1H NMR (400 MHz, CDCl3)δ4.86 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.61 (t, J=5.0 Hz, 2H), 2.69-2.68 (m, 2H), 2.57 (dt, J=15.0, 7.5 Hz, 4H), 2.32 (t, J=5.6 Hz, 2H), 2.28 (t, J=6.0 Hz, 2H), 1.67-1.49 (m, 14H), 1.42-1.33 (m, 2H), 1.28-1.25 (m, 34H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=6.8 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.3, 74.5, 64.3, 58.1, 56.1, 54.1, 53.7, 34.5, 34.4, 34.3, 32.0, 30.9, 29.7, 29.7, 29.5, 29.5, 29.4, 27.4 25.5, 25.1, 22.9, 22.8, 19.3, 14.2, 13.9.

[0266] Butyl 11-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)undecanoate (compound 35): [ka]

[0267] Compound 35 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (387 mg, 0.61 mmol, 91%). MS (ESI): m / z [M+H] + 641.1 for C 39 H 78 NO5; 1H NMR (400 MHz, CDCl3)δ4.86 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.54 (t, J=5.4 Hz, 2H), 2.59 (t, J=5.4 Hz, 2H), 2.53-2.42 (m, 4H), 2.29 (q, J=7.4 Hz, 4H), 1.66-1.55 (m, 6H), 1.54-1.46 (m, 6H), 1.44-1.33 (m, 4H), 1.32-1.19 (m, 36H), 0.92 (t, J=7.4 Hz, 3H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3)δ174.1, 173.3, 74.4, 64.5, 58.2, 55.9, 54.0, 53.6, 34.5, 34.4, 34.2, 31.9, 29.6, 29.6, 29.5, 29.3, 29.2, 28.4, 28.2, 27.4, 25.4, 25.1, 22.9, 22.7, 14.2, 14.0.

[0268] Butyl 11-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)undecanoate (compound 36): [ka]

[0269] Compound 36 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (331 mg, 0.53 mmol, 80%). MS (ESI): m / z [M+H] + 627.1 for C 38 H 76 NO5; 1H NMR (400 MHz, CDCl3)δ4.86 (qu, J=6.3 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.58 (t, J=5.2 Hz, 2H), 2.65 (t, J=5.1 Hz, 2H), 2.59-2.47 (m, 4H), 2.29 (dt, J=12.8, 7.5 Hz, 4H), 1.81 (qu, J=7.2 Hz, 2H), 1.64-1.54 (m, 4H), 1.55-1.45 (m, 6H), 1.43-1.33 (m, 2H), 1.33-1.19 (m, 36H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3)δ174.1, 173.2, 74.6, 64.2, 58.4, 55.9, 54.0, 53.9, 53.2, 34.5, 34.2, 32.2, 31.9, 30.8, 29.6, 29.6, 29.6, 29.3, 29.2, 27.4, 25.4, 25.1, 22.8, 19.2, 14.2, 13.8.

[0270] Butyl 12-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)dodecanoate (compound 37): [ka]

[0271] Compound 37 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (137 mg, 0.209 mmol, 33%). MS (ESI): m / z [M+H] + 655.0 for C 40 H 80 NO5; 1H NMR (400 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (qu, J=6.2 Hz, 1H), 4.08 (t, J=6.6 Hz, 2H), 3.61 (t, J=5.9 Hz, 2H), 2.68-2.60 (m, 2H), 2.57-2.47 (m, 4H), 2.33 (dt, J=13.1, 7.4 Hz, 4H), 1.67-1.58 (m, 6H), 1.57-1.47 (m, 8H), 1.46-1.36 (m, 4H), 1.35-1.20 (m, 37H), 0.95 (t, J=7.4 Hz, 3H), 0.89 (t, J=6.8 Hz, 6H); 13 C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 174.5, 173.7, 74.5, 64.1, 58.5, 55.4, 53.9, 53.5, 34.2, 34.1, 33.8, 31.6, 30.4, 29.3, 29.3, 29.3, 29.2, 29.2, 29.2, 29.0, 28.9, 28.8, 27.2, 26.2, 25.7, 25.1, 24.7, 22.7, 22.4, 18.8, 13.6, 13.2.

[0272] Butyl 12-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)dodecanoate (compound 38): [ka]

[0273] Compound 38 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (260 mg, 0.406 mmol, 64%). MS (ESI): m / z [M+H] + 641.0 for C 39 H 78 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.88 (qu, J=6.2 Hz, 1H), 4.08 (t, J=6.6 Hz, 2H), 3.65 (t, J=4.8 Hz, 2H), 2.79 - 2.70 (m, 2H), 2.69-2.55 (m, 4H), 2.36 (t, J=7.1 Hz, 2H), 2.31 (t, J=7.5 Hz, 2H), 1.88-1.79 (m, 2H), 1.66-1.59 (m, 4H), 1.58-1.46 (m, 6H), 1.44-1.36 (m, 2H), 1.35-1.23 (m, 38H), 0.95 (t, J=7.4 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 174.5, 173.6, 74.6, 64.0, 58.6, 55.5, 54.0, 53.1, 34.1, 33.8, 32.0, 31.6, 30.4, 29.3, 29.3, 29.2, 29.2, 29.2, 29.1, 28.9, 28.8, 27.2, 26.3, 25.0, 24.7, 22.3, 21.8, 18.8, 13.6, 13.2.

[0274] Butyl 15-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)pentadecanoate (compound 39): [ka]

[0275] Compound 39 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (193 mg, 0.28 mmol, 83%). MS (ESI): m / z [M+H] + 697.3 for C 43 H 85 NO5; 1H NMR (600 MHz, CDCl3)δ4.86 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.62-3.57 (m, 2H), 2.69-2.63 (m, 2H), 2.60-2.50 (m, 4H), 2.29 (dt, J=16.8, 7.4 Hz, 4H), 1.66-1.57 (m, 6H), 1.57-1.45 (m, 8H), 1.42-1.33 (m, 2H), 1.32-1.20 (m, 44H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.1, 173.3, 74.4, 64.2, 58.2, 55.9, 54.0, 53.6, 34.5, 34.4, 34.2, 31.9, 30.8, 29.7, 29.7, 29.6, 29.6, 29.5, 29.3, 29.3, 29.2, 27.4, 25.4, 25.1, 22.9, 22.7, 19.2, 14.2, 13.8.

[0276] Butyl 15-((4-(heptadecan-9-yloxy)-4-oxobutyl)(2-hydroxyethyl)amino)pentadecanoate (compound 40): [ka]

[0277] Compound 40 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (299 mg, 0.44 mmol, 78%). MS (ESI): m / z [M+H] + 683.2 for C 42 H 84 NO5; 1H NMR (600 MHz, CDCl3)δ4.86 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.58 (t, J=6.0 Hz, 2H), 2.68-2.61 (m, 2H), 2.60-2.47 (m, 4H), 2.30 (dt, J=19.5, 7.4 Hz, 4H), 1.85-1.77 (m, 2H), 1.64-1.57 (m, 4H), 1.55-1.43 (m, 6H), 1.41-1.33 (m, 2H), 1.32-1.19 (m, 44H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3)δ174.1, 173.2, 74.6, 64.2, 58.3, 56.0, 54.0, 53.2, 34.5, 34.2, 32.2, 31.9, 30.8, 29.7, 29.7, 29.6, 29.6, 29.6, 29.3, 29.3, 29.2, 27.5, 25.4, 25.1, 22.7, 19.2, 14.2, 13.8.

[0278] Pentyl 9-((2-hydroxyethyl)(5-oxo-5-(pentadecan-8-yloxy)pentyl)amino)nonanoate (compound 41): [ka]

[0279] Compound 41 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (278 mg, 0.465 mmol, 67%). MS (ESI): m / z [M+H] + 599.1 for C 36 H 72 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.91-4.83 (m, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.60 (t, J=5.9 Hz, 2H), 2.64 (t, J=5.9 Hz, 2H), 2.57-2.47 (m, 4H), 2.33 (dt, J=17.4, 7.4 Hz, 4H), 1.67-1.59 (m, 6H), 1.58-1.49 (m, 8H), 1.49-1.44 (m, 2H), 1.37-1.25 (m, 30H), 0.92 (t, J=7.0 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 174.4, 173.7, 74.4, 64.3, 58.5, 55.4, 53.9, 53.5, 34.2, 34.1, 33.8, 31.5, 29.2, 29.1, 29.0, 28.9, 28.8, 28.0, 27.8, 27.2, 26.3, 25.7, 25.0, 24.7, 22.7, 22.3, 22.0, 13.6, 13.4.

[0280] Pentyl 9-((2-hydroxyethyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)nonanoate (compound 42): [ka]

[0281] Compound 42 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (295 mg, 0.451 mmol, 65%). MS (ESI): m / z [M+H] + 655.1 for C 40 H 80 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.87 (qu, J=6.3 Hz, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.60 (t, J=5.9 Hz, 2H), 2.64 (t, J=5.9 Hz, 2H), 2.57-2.47 (m, 4H), 2.33 (dt, J=17.0, 7.4 Hz, 4H), 1.67-1.58 (m, 6H), 1.56-1.49 (m, 8H), 1.48-1.43 (m, 2H), 1.38-1.22 (m, 38H), 0.92 (t, J=7.0 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 174.4, 173.7, 74.5, 64.4, 58.5, 55.4, 53.9, 53.5, 34.2, 34.1, 33.8, 31.6, 29.2, 29.2, 29.1, 29.0, 29.0, 28.8, 28.0, 27.8, 27.2, 26.3, 25.7, 25.0, 24.7, 22.7, 22.4, 22.0, 13.6, 13.5.

[0282] Butyl 10-((2-hydroxyethyl)(5-oxo-5-(pentadecan-8-yloxy)pentyl)amino)decanoate (compound 43): [ka]

[0283] Compound 43 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (206 mg, 0.341 mmol, 53%). MS (ESI): m / z [M+H] + 598.2 for C 36 H 72 NO5; 1H NMR (400 MHz, CDCl3) δ 4.86 (qu, J=6.3 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.58 (t, J=4.8 Hz, 2H), 2.64 (m, 2H), 2.56-2.49 (m, 4H), 2.33-2.27 (m, 4H), 1.66-1.56 (m, 6H), 1.51-1.50 (m, 8H), 1.34-1.26 (m, 34H), 0.92-0.86 (m, 9H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.4, 74.5, 64.5, 58.3, 55.9, 54.0, 53.7, 34.5, 34.5, 34.3, 31.9, 29.6, 29.6, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 27.5, 25.5, 25.1, 23.0, 22.8, 22.5, 14.2, 14.1.

[0284] Pentyl 10-((2-hydroxyethyl)(5-oxo-5-(pentadecan-8-yloxy)pentyl)amino)decanoate (compound 44): [ka]

[0285] Compound 44 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (174 mg, 0.281 mmol, 44%). MS (ESI): m / z [M+H] + 612.2 for C 37 H 74 NO5; 1H NMR (400 MHz, CDCl3) δ 4.86 (p, J=6.3 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.56 (t, J=5.2 Hz, 2H), 2.61 (t, J=5.2 Hz, 2H), 2.53-2.46 (m, 4H), 2.32-2.26 (m, 4H), 1.66-1.57 (m, 6H), 1.54-1.33 (m, 10H), 1.27-1.26 (m, 31H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=6.8 Hz, 6H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.4, 74.5, 64.2, 58.4, 55.8, 54.0, 53.6, 34.5, 34.5, 31.9, 30.9, 29.6, 29.6, 29.4, 29.4, 29.3, 27.5, 27.1, 27.0, 26.5, 25.5, 25.1, 23.1, 22.8, 19.3, 14.2, 13.9.

[0286] Pentyl 10-((2-hydroxyethyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 45): [ka]

[0287] Compound 45 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as (mg, mmol, %).

[0288] Pentyl 9-((2-hydroxyethyl)(4-(nonadecan-10-yloxy)-4-oxobutyl)amino)nonanoate (compound 46): [ka]

[0289] Compound 46 was synthesized according to Synthetic Scheme 3 and general methods A, B, C, and G. The product was obtained as a pale yellow oil (100 mg, 0.155 mmol, 24%). MS (ESI): m / z [M+H] + 640.3 for C 39 H 78 NO5; 1 H NMR (400 MHz, CDCl3) δ 4.86 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.62 (t, J=5.2 Hz, 2H), 2.70 (t, J=5.1 Hz, 2H), 2.63-2.54 (m, 4H), 2.34-2.26 (m, 4H), 1.84 (p, J=7.2 Hz, 2H), 1.65-1.57 (m, 4H), 1.51-1.50 (m, 4H), 1.34-1.25 (m, 41H), 0.92-0.85 (m, 9H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.2, 74.8, 64.5, 58.2, 56.1, 54.1, 53.3, 34.5, 34.2, 32.1, 32.0, 29.7, 29.4, 29.4, 29.4, 29.2, 28.5, 28.2, 27.4, 26.4, 25.5, 25.1, 22.8, 22.5, 21.9, 14.2, 14.1.

[0290] Pentyl 10-((2-hydroxyethyl)(4-(nonadecan-10-yloxy)-4-oxobutyl)amino)decanoate (compound 47): [ka]

[0291] Compound 47 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (210 mg, 0.318 mmol, 49%). MS (ESI): m / z [M+H] + 654.6 for C 40 H 80 NO5;1 H NMR (400 MHz, CDCl3) δ 4.86 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.60 (bs, 2H), 2.68 (bs, 2H), 2.59-2.54 (m, 4H), 2.34-2.27 (m, 4H), 1.85-1.81 (m, 2H), 1.65-1.58 (m, 4H), 1.51-1.50 (m, 6H), 1.34-1.25 (m, 43H), 0.93-0.86 (m, 9H); 13 C NMR (100 MHz, CDCl3) δ 174.1, 173.2, 74.8, 64.5, 58.3, 56.2, 54.1, 53.3, 34.5, 34.2, 32.2, 32.0, 29.7, 29.6, 29.6, 29.5, 29.4, 29.3, 28.5, 28.2, 27.5, 25.5, 25.1, 22.8, 22.5, 14.3, 14.1.

[0292] Pentan-3-yl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 48) (also referred to as (I)(d)): [ka]

[0293] Compound 48 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (365 mg, 0.583 mmol, 56%). MS (ESI): m / z [M+H] + 627.0 for C 38 H 76 NO5; 1H NMR (600 MHz, CDCl3) δ 4.56 (qu, J=6.2 Hz, 1H), 4.78-4.72 (m, 1H)p, 3.51 (t, J=5.4 Hz, 2H), 2.56 (t, J = 5.4 Hz, 2H), 2.46 (dd, J=15.2, 7.Hz, 2H), 2.42 (dd, J=15.4, 7.8 Hz, 2H), 2.29 (td, J=7.5, 4.9 Hz, 4H), 1.66-1.59 (m, 4H), 1.59-1.52 (m, 4H), 1.52-1.45 (m, 7H), 1.45-1.38 (m, 2H), 1.35-1.18 (m, 38H), 0.87 (t, J = 7.5 Hz, 12H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.5, 76.5, 74.4, 58.5, 55.7, 54.0, 53.6, 34.8, 34.6, 34.3, 32.0, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 27.5, 27.3, 26.8, 26.6, 25.5, 25.3, 23.1, 22.8, 14.2, 9.7.

[0294] Isopentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 49): [ka]

[0295] Compound 49 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a pale yellow oil (398 mg, 0.636 mmol, 61%). MS (ESI): m / z [M+H] + 627.0 for C 38 H 76 NO5; 1H NMR (600 MHz, CDCl3) δ 4.86 (qu, J=6.2 Hz, 1H), 4.08 (t, J=6.9 Hz, 1H), 3.51 (t, J=5.4 Hz, 2H), 2.56 (t, J=5.4 Hz, 2H), 2.46 (dd, J=13.3, 5.9 Hz, 2H), 2.44-2.39 (m, 2H), 2.28 (dt, J=11.7, 7.5 Hz, 4H), 1.66-1.59 (m, 4H), 1.59-1.52 (m, 4H), 1.52-1.45 (m, 8H), 1.45-1.38 (m, 2H), 1.35-1.18 (m, 38H), 0.91 (d, J=6.7 Hz, 6H), 0.87 (t, J=7.5 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.0, 173.4, 74.4, 63.0, 58.5, 55.7, 53.9, 53.6, 37.5, 34.6, 34.5, 34.3, 32.0, 30.0, 29.6, 29.5, 29.4, 29.4, 29.3, 27.5, 27.3, 26.8, 25.5, 25.2, 25.1, 23.1, 22.8, 22.6, 14.2.

[0296] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxypropyl)amino)nonanoate (compound 50): [ka]

[0297] Compound 50 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (410 mg, 0.641 mmol, 64%). MS (ESI): m / z [M+H] + 641.3 for C 39 H 78 NO5; 1H-NMR (600 MHz, CDCl3) δ 4.87 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.71 (n, J=3.0 Hz, 1H), 3.59 (bs, 1H), 2.56-2.48 (m, 2H), 2.43-2.38 (m, 2H), 2.36 (dd, J=12.5, 2.9 Hz, 1H), 2.29 (t, J=7.3 Hz, 4H), 2.21 (t, J=11.5 Hz, 1H), 1.66-1.57 (m, 6H), 1.51-1.48 (m, 8H), 1.36-1.26 (m, 36H), 1.11 (d, J=6.1 Hz, 3H), 0.91 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.3, 74.2, 64.463.0,62.4, 54.2, 53.9, 34.5, 34.4, 34.1 (2x), 31.8(2x), 29.5(2x), 29.5 (2x), 29.4, 29.3, 29.2(2x), 29.1, 28.3, 28.1, 27.4,27.2, 26.7, 25.3(2x), 25.0, 23.0, 22.6(2x), 22.3, 19.8, 14.1(2x), 13.9.

[0298] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)((2R)-2-hydroxypropyl)amino)nonanoate (compound 51): [ka]

[0299] Compound 51 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (150 mg, 0.234 mmol, 71%). MS (ESI): m / z [M+H] + 641.1 for C 39 H 78 NO5; [α] D25 = - 23,8 (c 6,15, MTBE); 1 H NMR (600 MHz, 4.87 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.71 (dtd, J=16.5, 6.1, 3.0 Hz, 1H), 3.59 (bs, 1H), 2.56-2.48 (m, 2H), 2.43-2.38 (m, 2H), 2.36 (dd, J=12.5, 2.9 Hz, 1H), 2.30 (t, J=7.3 Hz, 2H), 2.29 (t, J=7.3 Hz, 2H), 2.21 (dd, J=12.6, 10.5 Hz, 1H), 1.66-1.57 (m, 6H), 1.51-1.48 (m, 8H), 1.36-1.26 (m, 36H), 1.11 (d, J=6.1 Hz, 3H), 0.91 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.3, 74.2, 64.3, 63.0, 62.4, 54.2, 53.9, 34.5, 34.3, 34.1(2x), 31.8(2x), 29.5(2x), 29.5 (2x), 29.4, 29.2, 29.2(2x), 29.1, 28.3, 28.1, 27.4, 27.2, 26.7, 25.3(2x), 25.0, 23.0, 22.6(2x), 22.3, 19.8, 14.1(2x), 13.9.

[0300] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)((2S)-2-hydroxypropyl)amino)nonanoate (compound 52): [ka]

[0301] Compound 52 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (150 mg, 0.234 mmol, 71%). MS (ESI): m / z [M+H] + 641,1 for C 39 H 78 NO5; [α] D 25 = + 23,8(c 7,15, MTBE); 1 H NMR (600 MHz, CDCl3) δ 4.87 (qu, J=6.2 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.71 (dtd, J=16.5, 6.1, 3.0 Hz, 1H), 3.59 (bs, 1H), 2.56-2.48 (m, 2H), 2.43-2.38 (m, 2H), 2.36 (dd, J=12.5, 2.9 Hz, 1H), 2.30 (t, J=7.3 Hz, 2H), 2.29 (t, J=7.3 Hz, 2H), 2.21 (dd, J=12.6, 10.5 Hz, 1H), 1.66-1.57 (m, 6H), 1.51-1.48 (m, 8H), 1.36-1.26 (m, 36H), 1.11 (d, J=6.1 Hz, 3H), 0.91 (t, J=7.0 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.3, 74.2, 64.4, 63.0, 62.4, 54.2, 53.9, 34.5, 34.4, 34.1(2x), 31.8(2x), 29.5(2x), 29.5(2x), 29.4, 29.3, 29.2(2x), 29.1, 28.3, 28.1, 27.4, 27.2, 26.7, 25.3(2x), 25.0, 23.0, 22.6(2x), 22.3, 19.8, 14.1(2x), 13.9.

[0302] Pentyl 9-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (compound 53) (Method H): [ka]

[0303] A solution of pentyl 9-((4-((tert-butoxycarbonyl)amino)butyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (Intermediate E) (0.95 g, 1.26 mmol, 1 equiv) in dioxane (1 ml) was cooled to 0° C. and a 4 N solution of HCl in dioxane (11.0 ml, 44.1 mmol, 35 equiv) was added over 10 min. The reaction mixture was stirred at room temperature for 20 h. The solvent and volatiles were then evaporated to dryness. The product was purified by silica gel chromatography (Phase A: t-BuOMe; Phase B t-BuOMe:MeOH:NH4OH = 50:50:1; 100% A to A:B = 50:50) to give pentyl 9-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (700 mg, 1.07 mmol, 85%) as a white wax. MS (ESI): m / z [M+H] + 654,0 for C 40 H 81 N2O4; 1 H NMR (600 MHz, CDCl3) δ 6.08 (bs, 2H), 4.85 (p, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 2.93 (t, J=6.1 Hz, 2H), 2.60-2.60 (m, 6H, 3xCH2N), 2.34 (t, J=7.2 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.82 (p, J=6.2 Hz, 2H), 1.71 (p, J=6.3 Hz, 2H), 1.61 (m, 8H), 1.51 (m, 6H), 1.26-1.26 (m, 36H), 0.91 (t, J=7.1 Hz, 3H), 0.88 (t, J=7.0 Hz, 6H); 13C NMR (150 MHz, CDCl3) δ 174.0, 173.2, 74.5, 64.4, 53.3, 52.8, 52.3, 40.0, 34.3, 34.2, 34.1, 31.8, 29.5, 29.5, 29.2, 29.2, 29.2, 29.0, 28.3, 28.1, 27.5, 27.4, 25.3, 24.9, 24.8, 24.6, 24.4, 23.0, 22.6, 22.3, 14.1(2x), 13.9.

[0304] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)amino)nonanoate (compound 54) (Method I): [ka]

[0305] To a solution of pentyl 9-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (compound 53) (300 mg, 0.459 mmol, 1 equiv.) in ethanol (10 mL), 3-methoxy-4-(methylamino)cyclobut-3-ene-1,2-dione (intermediate F) (79 mg, 0.505 mmol, 1.1 equiv.) was added. The reaction mixture was stirred at 40 °C for 20 h. The solvent was evaporated to dryness, and the crude product was purified by silica gel chromatography (Phase A: t-BuOMe; Phase B t-BuOMe:MeOH:NH4OH = 50:50:1; 100% A to A:B = 50:50) to give pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)amino)nonanoate (compound 54) (220 mg, 0.289 mmol, 63%) as a yellow wax. MS (ESI): m / z [M+H] + 763,2 for C 45 H 84 N3O6; 1H NMR (600 MHz, benzene-d6) δ 8.47 (bs, 1), 8.16 (bs, 1), 5.19-5.14 (m, 1H), 4.08 (t, J=6.7 Hz, 2H), 3.86 (bs, 2H), 3.74 (bs, 1 H), 3.32 (d, 2.50 (t, J=6.8 Hz, 2H), 2.43 (t, J=7.2 Hz, 4H), 2.37 (t, J=7.4 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H), 1.85-1.85 (m, 2H), 1.77-1.77 (m, 2H), 1.67-1.67 (m, 6H), 1.54-1.54 (m, 8H), 1.33-1.33 (m, 32H), 1.23-1.23 (m, 4H), 0.97 (t, J=7.1 Hz, 6H), 0.87 (t, J=7.1 Hz, 3H); 13 C NMR (150 MHz, benzene-d6+CD3OD) δ 183.8, 183.5, 174.7, 174.7, 174.3, 174.3, 169.3, 169.0, 128.7, 75.1, 65.1, 60.3, 54.7, 54.2, 44.7, 35.1, 35.0, 34.9, 32.7, 32.7, 31.0, 30.3, 30.3, 30.3, 30.2, 30.1, 30.1, 30.1, 29.8, 29.8, 29.1, 28.8, 28.3, 27.2, 26.6, 26.2, 25.7, 24.4, 23.9, 23.4, 23.0, 14.6, 14.5, 14.3.

[0306] Pentyl 9-(3-aminopropyl-(5-(1-octylnonoxy)-5-oxo-pentyl)amino)nonanoate (compound 55): [ka]

[0307] Compound 55 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (470 mg, 0.735 mmol, 86%). MS (ESI): m / z [M+H] + 640,1 for C 39 H 79 N2O4; 1 H NMR (600 MHz, CDCl3) δ 4.86 (qu, 1H, J=6.3, 1.0 Hz), 4.06 (t, J=6.8 Hz, 2H), 2.74 (t, J=6.8 Hz, 2H), 2.45 (t, J=7.1 Hz, 2H), 2.41 (t, J=7.5 Hz, 2H), 2.37 (t, J=7.6 Hz, 2H), 2.29 (t, 7.2 Hz, 2H), 2.28 (t, J=7.2 Hz, 4H), 1.78 (s, 2H), 1.55-1.68 (m, 8H), 1.37-1.55 (m, 8H), 1.17-1.37 (m, 36H), 0.91 (t, J=7.1 Hz, 3H), 0.88 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.4, 74.2, 64.4, 54.1, 53.7, 52.0, 40.9, 34.6, 34.4, 34.1, 31.8, 30.7, 29.5, 29.5, 29.4, 29.3, 29.2, 29.1, 28.3, 28.1, 27.5, 27.0, 26.5, 25.3, 25.0, 23.1, 22.6, 22.3, 14.1, 13.9.

[0308] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)nonanoate (compound 56): [ka]

[0309] Compound 56 was synthesized according to Representative Procedure 3 and General Method I. The product was obtained as a yellow wax (250 mg, 0.334 mmol, 97%). MS (ESI): m / z [M+H] + 749,2 for C 44 H 82 N3O6; 1 H NMR (400 MHz, benzene-d6) δ 8.40 (bs, 1H), 8.10 (bs, 1H), 5.20-5.10 (m, 1H), 4.08 (t, J=6.7 Hz, 2H), 3.82 (bs, 2H), 3.32 (d, J=4.7 Hz, 2H), 2.59 (t, J=6.2 Hz, 2H), 2.45 (t, J=7.0 Hz, 4H), 2.36 (t, J=7.3 Hz, 2H), 2.28 (t, J=7.5 Hz, 2H), 1.95 (dd, J=5.4 Hz, 2H), 1.13-1.84 (m, 50H), 0.96 (t, J=6.8 Hz, 6H), 0.86 (t, J=7.0 Hz, 3H); 13 C NMR (100 MHz, benzene-d6) δ 183.4, 183.0, 174.3, 173.9, 168.8, 168.5, 74.6, 64.6, 54.1, 53.7, 51.3, 42.9, 34.6, 34.5, 34.4, 32.2(2x), 30.6, 29.9(2x),29.9(2x), 29.8, 29.6, 29.6(2x), 29.4, 28.6(2x), 28.6, 28.3(2x), 27.9, 26.7, 26.2, 25.8(2x), 25.3, 23.4, 23.0(2x), 22.6, 14.1(2x), 13.9.

[0310] Pentyl 9-((3-aminopropyl)(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)nonanoate (compound 57): [ka]

[0311] Compound 57 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (1140 mg, 1.82 mmol, 83%). MS (ESI): m / z [M+H] + 626,0 for C 38 H 77 N2O4; 1 H NMR (600 MHz, CDCl3) δ 4.86 (qu, J=6.3, 1.0 Hz, 1H), 4.06 (t, J=6.8 Hz, 2H), 2.73 (t, J=6.8 Hz, 2H), 2.45 (t, J=7.1 Hz, 2H), 2.42 (t, J=7.3 Hz, 2H), 2.38 (t, J=7.6 Hz, 2H), 2.28 (t, J=7.7 Hz, 1H), 2.27 (t, J=7.7 Hz, 1H), 1.83 (bs, 2H), 1.74 (q, J=7.4, 2H), 1.65-1.56 (m, 6H), 1.51-1.50 (m, 4H), 1.43-1.38 (m, 2H), 1.35-1.26 (m, 36H), 0.89 (t, J=7.1, 3H), 0.86 (t, J=7.1, 6H); 13 C NMR (150 MHz, CDCl3) δ 173.9, 173.5, 74.2, 64.3, 54.1, 53.3, 51.9, 50.5, 40.7, 34.3, 34.1(2x), 32.4, 31.8(2x), 30.8, 29.5(2x), 29.5(2x), 29.4, 29.3, 29.2(2x), 29.1, 28.3, 28.0, 27.5, 27.0, 25.3(2x), 25.0, 22.6(2x), 22.6, 22.3, 14.0(2x), 13.9.

[0312] Pentyl 9-((4-(heptadecan-9-yloxy)-4-oxobutyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)nonanoate (compound 58): [ka]

[0313] Compound 58 was synthesized according to Representative Procedure 3 and General Method I. The product was obtained as a yellow wax (345 mg, 0.470 mmol, 98%). MS (ESI): m / z [M+H] + 735.1 for C 43 H 80 N3O6; 1 H NMR (600 MHz, CDCl3) δ 7.21 (bs, 1H), 6.76 (bs, 1H), 4.82 (qu, J = 6.3 Hz, 1H), 4.05 (t, J = 6.8 Hz, 2H), 3.72 (bs, 2H), 3.31 (d, J = 5.0 Hz, 3H), 2.52 (t, J = 6.3 Hz, 2H), 2.42 (t, J = 6.9 Hz, 2H), 2.39 (t, J = 7.7 Hz, 2H), 2.36 (t, J = 7.3 Hz, 2H), 2.29 (t, J = 7.5 Hz, 2H), 1.79 - 1.74 (m, 4H), 1.65 - 1.59 (m, 4H), 1.53 - 1.52 (m, 4H), 1.41 - 1.26 (m, 38H), 0.91 (t, J = 7.1 Hz, 3H), 0.88 (t, J = 7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3 - CD3OD = 2:1) δ 183.4, 182.9, 175.2, 174.6, 169.0, 168.7, 75.4, 65.1, 54.4, 53.5, 51.5, 43.2, 34.8, 34.6 (2x), 32.9, 32.3 (2x), 31.2, 30.0 (2x), 29.9, 29.7 (2x), 29.7, 29.5, 28.8, 28.6, 28.0, 26.9, 25.8 (2x), 25.4, 23.1 (2x), 22.7, 22.5, 14.3 (2x), 14.1.

[0314] Pentyl 6-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)hexanoate (compound 59): [ka]

[0315] Compound 59 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (590 mg, 0.827 mmol, 93%). MS (ESI): m / z [M+H] + 707.1 for C 41 H 76 N3O6; 1 H NMR (600 MHz, CDCl3-CD3OD=2:1)δ4.89-4.85 (m, 1H), 4.08 (t, J=6.8 Hz, 2H), 3.70 (t, J=6.3 Hz, 2H), 3.30 (bs, 3H), 3.16 (bs, 2H), 3.07 (bs, 4H), 2.41 (t, J=7.1 Hz, 2H), 2.36 (t, J=7.3 Hz, 2H), 2.06-2.01 (m, 2H), 1.76-1.62 (m, 10H), 1.55-1.53 ​​(m, 4H), 1.44-1.38 (m, 2H), 1.36-1.33 (m, 4H), 1.32-1.25 (m, 26H), 0.92 (t, J=7.1 Hz, 3H), 0.89 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 183.0, 182.9, 174.3, 173.6, 169.8, 167.9, 75.5, 65.2, 53.2, 53.0, 51.1, 41.2, 34.4, 34.1, 33.9, 32.2, 31.1, 29.8, 29.8, 29.6, 28.6, 28.4, 26.4, 26.2, 25.7, 24.6, 23.8, 23.5, 23.0, 22.6, 22.3, 14.2, 14.1.

[0316] Pentyl 6-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)amino)hexanoate (compound 60): [ka]

[0317] Compound 60 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (521 mg, 0.716 mmol, 100%). MS (ESI): m / z [M+H] + 721.2 for C 42 H 78 N3O6; 1 H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.90-4.86 (m, 1H), 4.08 (t, J=6.8 Hz, 2H), 3.67 (t, J=6.4 Hz, 2H), 3.29 (bs, 3H), 3.12-3.04 (m, 6H), 2.41 (t, J=7.1 Hz, 2H), 2.37 (t, J=7.3 Hz, 2H), 1.84-1.79 (m, 2H), 1.77-1.62 (m, 12H), 1.55-1.53 ​​(m, 4H), 1.44-1.39 (m, 2H), 1.38-1.33 (m, 4H), 1.33-1.25 (m, 26H), 0.92 (t, J=7.1 Hz, 3H), 0.89 (t, J=7.1 Hz, 6H); 13C NMR (150 MHz, CDCl3-CD3OD=2:1) ​​δ 182.9, 182.8, 174.3, 173.6, 169.6, 168.2, 75.5, 65.2, 53.2, 53.1, 53.0, 43.0, 34.4, 34.1, 33.9, 32.2, 31.0, 29.8, 29.8, 29.6, 28.6, 28.4, 28.2, 26.4, 25.7, 24.6, 23.8, 23.5, 23.0, 22.6, 22.3, 21.2, 14.2, 14.1;

[0318] Butyl 11-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)undecanoate (compound 61): [ka]

[0319] Compound 61 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a white solid (230 mg, 0.30 mmol, 45%). MS (ESI): m / z [M+H] + 763.3 for C 45 H 84 N3O6; 1H NMR (400 MHz, CDCl3)δ4.81 (p, J=6.3 Hz, 1H), 4.06 (t, J=6.7 Hz, 2H), 3.80-3.65 (m, 2H), 3.29 (d, J=5.1 Hz, 3H), 2.53 (t, J=5.3 Hz, 2H), 2.45 (t, J=6.7 Hz, 2H), 2.38 (dt, J=14.0, 7.1 Hz, 4H), 2.28 (t, J=6.7 Hz, 4H), 1.81-1.72 (m, 2H), 1.68-1.57 (m, 6H), 1.56-1.45 (m, 6H), 1.44-1.35 (m, 4H), 1.33-1.19 (m, 36H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 182.8, 182.7, 174.0, 169.0, 167.9, 74.9, 64.2, 53.4, 53.1, 34.4, 34.1, 31.9, 31.3, 29.6, 29.6, 29.6, 29.5, 29.4, 29.3, 29.3, 29.2, 28.4, 28.4, 27.3, 25.4, 25.1, 22.7, 19.2, 14.2, 13.8.

[0320] Pentyl 11-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)undecanoate (compound 62): [ka]

[0321] Compound 62 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow solid (349 mg, 0.45 mmol, 94%). MS (ESI): m / z [M+H] + 777.3 for C 46 H 86 N3O6;1 H NMR (400 MHz, CDCl3)δ4.82 (p, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.78-3.65 (m, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.62 (t, J=5.8 Hz, 2H), 2.57-2.43 (m, 4H), 2.34 (t, J=7.0 Hz, 2H), 2.28 (t, J=7.5 Hz, 2H), 1.84-1.78 (m, 2H), 1.69-1.57 (m, 6H), 1.54-1.40 (m, 8H), 1.33-1.18 (m, 40H), 0.90 (t, J=5.9 Hz, 3H), 0.87 (t, J=6.7 Hz, 6H); 13 C NMR (100 MHz, CDCl3-CD3OD=2:1) ​​δ 183.3, 183.2, 174.1, 168.8, 168.1, 75.0, 64.5, 53.4, 53.2, 34.5, 34.2, 34.1, 31.9, 31.3, 29.6, 29.6, 29.6, 29.5, 29.3, 29.3, 29.2, 28.4, 28.4, 28.2, 27.6, 25.8, 25.4, 25.1, 22.9, 22.7, 22.4, 20.2, 14.2, 14.0.

[0322] Butyl 12-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)dodecanoate (compound 63): [ka]

[0323] Compound 63 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (832 mg, 1.072 mmol, 70%). MS (ESI): m / z [M+H] + 776.7 for C 46 H 86N3O6; 1 H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.90-4.84 (m, 1H), 4.08 (t, J=6.7 Hz, 2H), 3.63 (bs, 2H), 3.28 (s, 3H), 2.55-2.51 (m, 2H), 2.51-2.47 (m, 2H), 2.47-2.42 (m, 2H), 2.35 (t, J=7.4 Hz, 2H), 2.31 (td, J=7.6, 3.4 Hz, 2H), 1.80-1.73 (m, 2H), 1.66-1.58 (m, 6H), 1.57-1.48 (m, 6H), 1.48-1.36 (m, 4H), 1.34-1.23 (m, 40H), 0.95 (t, J=7.4 Hz, 3H), 0.89 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3- CD3OD=2:1 ) δ 183.3, 182.8, 175.1, 174.5, 168.9, 168.5, 75.2, 64.7, 54.2, 53.8, 51.4, 43.1, 34.8, 34.7, 34.4, 32.2, 31.1, 31.0, 30.0, 29.9, 29.8, 29.8, 29.8, 29.6, 29.6, 29.4, 28.5, 28.0, 26.5, 26.0, 25.7, 25.3, 23.4, 23.0, 19.4, 14.2, 13.8.

[0324] Pentyl 12-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)dodecanoate (compound 64): [ka]

[0325] Compound 64 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (246 mg, 0.311 mmol, 85%). MS (ESI): m / z [M+H] + 790.6 for C 47 H 88 N3O6; 1 H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.80 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.74 (bs, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.53-2.47 (m, 2H), 2.43 (t, J=6.5 Hz, 2H), 2.36 (t, J=7.0 Hz, 4H), 2.28 (td, J=7.6, 2.8 Hz, 2H), 1.75 (dt, J=11.1, 5.6 Hz, 2H), 1.67-1.57 (m, 6H), 1.55-1.50 (m, 4H), 1.49-1.43 (m, 2H), 1.43-1.37 (m, 2H), 1.36-1.20 (m, 44H), 0.92-0.86 (m, 9H).

[0326] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)((4-((1H-imidazol-5-yl)formamido)butyl))amino)nonanoate (compound 65) (Method J): [ka]

[0327] A mixture of 4-imidazolecarboxylic acid (33 mg, 0.286 mmol, 1.1 equiv.), oxalyl chloride (0.57 mL, 6.51 mmol, 25 equiv.), and 1 drop of dimethylformamide was stirred at room temperature for 3 h. The volatiles were then evaporated to dryness, and the residue was added to dichloromethane (2 mL) and a solution of pentyl 9-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)nonanoate (compound 53) (170 mg, 0.26 mmol, 1.0 equiv.) in dichloromethane (2 mL) and DIPEA (0.227 mL, 1.30 mmol, 5.0 equiv.). The reaction mixture was stirred at room temperature for 20 h. Water (40 mL) and ethyl acetate (10 mL) were then added to the reaction mixture. The phases were separated, and the aqueous phase was extracted with ethyl acetate (2 × 20 mL). The combined organic phases were dried over sodium sulfate and evaporated to dryness to give a crude yellow oil (230 mg). The product was purified by silica gel chromatography (Phase A: t-BuOMe; Phase B: t-BuOMe:MeOH:NH4OH = 50:50:1; 100% to A:B = 50:50) to give pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)((4-((1H-imidazol-5-yl)formamido)butyl))amino)nonanoate (Compound 65) (160 mg, 0.214 mmol, 82%) as a yellow oil. MS (ESI): m / z [M+H] + 748,2 for C 44 H 83 N4O5; 1H NMR (600 MHz, CDCl3) δ 11.37 (bs, 1H), 7.58 (s, 1H), 7.57 (s, 1H), 7.28 (bs, 1H), 4.88-4.83 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.44 (dt, J=6.8, 6.3 Hz, 2H), 2.41 (t, J=7.9 Hz, 2H), 2.40 (t, J=7.9 Hz, 2H), 2.38-2.34 (m, 2H), 2.30-2.26 (m, 4H), 1.25-1.25 (m, 54H), 0.90 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.1, 173.5, 163.2, 136.7, 135.0, 118.7, 74.2, 64.4, 54.1, 53.7, 53.6, 38.9, 34.6, 34.4, 34.1, 31.8, 29.5, 29.5, 29.4, 29.3, 29.2, 29.1, 28.3, 28.1, 27.7, 27.6, 27.0, 26.5, 25.3, 25.0, 24.6, 23.2, 22.6, 22.3, 14.1, 13.9.

[0328] Pentyl 9-((4-(heptadecan-9-yloxy)-4-oxobutyl)((3-((1H-imidazol-4-yl)formamido)propyl))amino)nonanoate (compound 66): [ka]

[0329] Compound 66 was synthesized according to Representative Procedures 2 and 4, and General Methods A, C, D, E, H, and J. The product was obtained as a yellow tight oil (210 mg, 0.292 mmol, 91%). MS (ESI): m / z [M+H] + 720.2 for C 42 H 79 N4O5; 1H NMR (600 MHz, CDCl3) δ 11.3 (bs, 1H), 8.10 (s, 1H), 7.71 (bs, 1H), 7.58 (s, 1H), 7.56 (s, 0.9H), 4.88-4.83 (m, 1H), 4.30-4.25 (m, 1H), 4.06 (t, J=6.8 Hz, 2H), 3.48 (dt, J=6.3, 6.5 Hz, 2H), 2.53 (t, J=6.5 Hz, 2H), 2.45 (t, J=7.3 Hz, 2H), 2.40 (t, J=7.6 Hz, 2H), 2.32 (t, J=7.4 Hz, 2H), 2.28 (t, J=7.6 Hz, 2H), 1.78-1.78 (m, 4H), 1.61-1.61 (m, 4H), 1.50-1.42 (m, 6H), 1.33-1.33 (m, 4H), 1.25-1.25 (m, 32H), 0.91 (t, 3H), 0.86 (t, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.1, 173.5, 165.9, 135.1, 134.2, 129.5, 74.3, 67.8, 64.4, 54.0, 53.3, 52.3, 38.9, 34.4, 34.1, 32.4, 31.8, 30.6, 29.5, 29.5, 29.4, 29.2, 29.2, 29.1, 28.3, 28.1, 27.5, 25.3, 25.0, 24.0, 22.9, 22.6, 22.3, 14.1, 13.9.

[0330] Butyl 11-((5-(heptadecan-9-yloxy)-5-oxopentyl)((3-((1H-imidazol-4-yl)formamido)propyl))amino)undecanoate (compound 67): [ka]

[0331] Compound 67 was synthesized according to Representative Procedures 2 and 4, and General Methods A, C, D, E, H, and J. The product was obtained as a yellow tight oil (150 mg, 0.201 mmol, 66%). MS (ESI): m / z [M+H] + 748.2 for C 44 H 83 N4O5; 1 H NMR (600 MHz, CDCl3) δ 11.31 (bs, 1H), 7.73 (bs, 1H), 7.57 (bs, 1H), 7.54 (bs, 1H), 4.88-4.83 (m, 1H), 4.07 (t, J=6.7 Hz, 2H), 3.48 (dt, J=6.3, 6.3 Hz, 2H), 2.52 (t, J=6.2 Hz, 2H), 2.43 (t, J=7.3 Hz, 2H), 2.39 (t, J=7.5 Hz, 2H), 2.29 (t, J=7.5 Hz, 4H), 1.74 (q, J=6.7 Hz, 2H), 1.64-1.58 (m, 6H), 1.52-1.46 (m, 8H), 1.40-1.33 (m, 2H), 1.30-1.25 (m, 36H), 0.93 (t, J=7.4 Hz, 3H), 0.87 (t, J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3) δ 174.3, 173.6, 163.3, 136.9, 135.1, 118.7, 74.3, 64.2, 54.2, 53.8, 52.4, 38.2, 34.7, 34.5, 34.2, 31.9, 30.8, 29.7, 29.6, 29.6, 29.6, 29.5, 29.3, 29.2, 27.7, 27.0, 26.5, 25.4, 25.1, 23.3, 22.7, 19.2, 14.2, 13.8.

[0332] Pentyl 6-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)hexanoate (compound 68): [ka]

[0333] Compound 68 was synthesized according to Representative Procedure 1 using Intermediate G and general methods F, B, C, and G. The product was obtained as a pale yellow oil (70 mg, 0.114 mmol, 23%). MS (ESI): m / z [M+H] + 584.9 for C 35 H 70 NO5; 1 H NMR (600 MHz, CDCl3-CD3OD=2:1) ​​δ 4.90-4.85 (m, 1H), 4.08 (t, J=6.6 Hz, 2H), 3.65 (t, J=5.7 Hz, 2H), 2.74 (bs, 2H), 2.63 (bs, 3H), 2.36 (t, J=7.3 Hz, 2H), 2.36 (t, J=7.3 Hz, 2H), 1.70-1.61 (m, 6H), 1.59-1.52 (m, 8H), 1.40-1.34 (m, 2H), 1.34-1.25 (m, 29H), 0.91 (t, J=7.1 Hz, 3H), 0.89 (t, J=7.0 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1)δ175.0, 174.1, 75.3, 64.5, 55.7, 54.2, 54.0, 34.6, 34.5, 34.4, 32.2, 31.6, 29.8, 29.8, 29.6, 28.7, 25.7, 25.0, 24.2, 23.9, 23.0, 22.9, 14.2, 14.0.

[0334] Pentan-3-yl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)nonanoate (compound 69): [ka]

[0335] Compound 69 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (390 mg, 0.521 mmol, 70%). MS (ESI): m / z [M+H] + 748.9 for C 44 H 82 N3O6; 1 H NMR (600 MHz, CDCl3) δ 4.81 (qu, J=6.2 Hz, 1H), 4.74 (tt, J = 7.1, 5.3 Hz, 1H), 3.70 (bs, 2H), 3.29 (d, J = 5.0 Hz, 3H), 2.48 (t, J = 6.1 Hz, 2H), 2.41 (t, J = 6.9 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.33 (t, J=7.2 Hz, 2H), 2.28 (t, J = 7.5 Hz, 2H), 1.75 (qu, J = 6.2 Hz, 2H), 1.64-1.57 (m, 4H), 1.57-1.48 NMR (151 MHz, ) δ 183.4, 183.2, 174.0, 168.5, 168.3, 76.6, 75.0, 53.8, 53.6, 34.8, 34.8, 34.5, 34.2, 32.0, 32.0, 32.0, 31.3, 29.7, 29.7, 29.7, 29.6, 29.6, 29.6, 29.6, 29.4, 29.4, 29.4, 29.4, 29.3, 28.1, 27.7, 26.6, 26.6, 26.6, 26.6, 26.6, 26.6, 26.6, 25.5, 25.3, 23.1, 22.8, 22.8, 14.2, 14.2, 9.8, 9.7, 9.7, 9.7.

[0336] Isopentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)nonanoate (compound 70): [ka]

[0337] Compound 70 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (550 mg, 0.735 mmol, 71%). MS (ESI): m / z [M+H] + 749.0 for C 44 H 82 N3O6; 1 H NMR (600 MHz, CDCl3+CD3OD) δ 4.76 (qu, J=6.3 Hz, 1H), 4.01 (t, J=6.9 Hz, 2H), 3.59 (bs, 1H), 3.47 (bs, 1H), 3.20 (s, 3H), 3.00 (q, J=7.3 Hz, 3H), 2.88-2.80 (m, 2H), 2.79-2.65 (m, 2H), 2.30-2.24 (m, 2H), 2.21 (t, J=7.5 Hz, 2H), 1.90-1.80 (m, 2H), 1.65-1.45 (m, 10H), 1.45-1.39 (m, 7H), 1.30-1.10 (m, 42H), 0.83 (t, J=3.7 Hz, 6H), 0.79 (t, J = 7.1 Hz, 6H); 13C NMR (150 MHz, CDCl3+CD3OD) δ 182.6, 174.0, 173.2, 169.1, 168.0, 162.4, 162.1, 117.9, 115.9, 74.8, 63.0, 63.0, 53.2, 52.9, 51.1, 46.1, 37.5, 34.4, 34.4, 34.4, 34.4, 34.4, 34.1, 34.0, 32.0, 32.0, 32.0, 31.2, 29.7, 29.6, 29.6, 29.6, 29.6, 29.4, 29.4, 29.3, 29.3, 29.3, 29.3, 29.2, 29.2, 29.1, 29.0, 27.1, 25.4, 25.2, 25.0, 25.0, 22.8, 22.8, 22.8, 22.8, 22.6, 22.6, 22.6, 22.5, 22.5, 14.2, 14.2, 9.1.

[0338] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-hydroxypropyl)amino)nonanoate (compound 71): [ka]

[0339] Compound 71 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (560 mg, 0.875 mmol, 80%). MS (ESI): m / z [M+H] + 640.5 for C 39 H 77NO5; 1H-NMR (400 MHz, CDCl3) δ 5.47 (bs, 1H), 4.85 (qu, J=6.3 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 3.77 (t, J=5.1 Hz, 2H), 2.61 (t, J=5.6 Hz, 2H), 2.47-2.36 (m, 4H), 2.31 (t, J=7.5 Hz, 2H), 2.29 (t, 7.3 Hz, 2H), 1.70-1.57 (m, 8H), 1.56-1.40 (m, 8H), 1.38-1.20 (m, 36H), 0.91 (t, J=7.0 Hz, 3H), 0.88 (t, J=6.9 Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.2, 74.2, 64.7, 64.3, 55.1, 54.2, 53.7, 34.4, 34.3, 34.1, 31.8, 29.5, 29.5, 29.4, 29.2, 29.1, 28.3, 28.1, 27.8, 27.4, 26.8, 26.2, 25.3, 24.9, 23.0, 22.6, 22.3, 14.1, 13.9

[0340] Pentyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-hydroxybutyl)amino)nonanoate (compound 72): [ka]

[0341] Compound 72 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (340 mg, 0.520 mmol, 75%). MS (ESI): m / z [M+H] + 654.6 for C 40 H 79 NO5; 1H NMR (400 MHz, CDCl3) δ 6.27 (bs, 1H), 4.85 (qu,J=6.3 Hz,1H), 4.04 (t, J=6.8 Hz, 2H), 3.53 (t, J=4.7 Hz, 2H), 2.46-2.36 (m, 6H), 2.28 (t, J=7.8, 2H), 2.26 (t, J=7.4 Hz, 2H), 1.65-1.53 ​​(m, 10H), 1.53-1.38 (m, 8H), 1.33-1.20 (m, 36H), 0.88 (t, J=7.0, 3H), 0.85 (t, J=6.9Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.3, 74.2, 64.3, 62.7, 54.6, 53.7, 53.2, 34.4, 34.3, 34.1, 32.7, 31.8, 29.5, 29.5, 29.3, 29.2, 29.2, 29.1, 28.3, 28.1, 27.6, 26.3, 26.0, 25.4, 25.3, 25.0, 23.2, 22.6, 22.3, 14.1, 13.9.

[0342] 2-Methylbutyl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 73): [ka]

[0343] Compound 73 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (510 mg, 0.815 mmol, 78%). MS (ESI): m / z [M+H] + 627.3 for C 38 H 76 NO5; 1H NMR (600 MHz, CDCl3) δ 4.89-4.82 (m, 1H), 3.95 (dd, J=10.7, 6.0 Hz, 1H), 3.88-3.83 (m, 1H), 3.52 (t, J=5.4 Hz, 2H), 2.56 (t, J=5.4 Hz, 2H), 2.44 (ddd, J=25.1, 16.2, 8.7 Hz, 4H), 2.29 (td, J=7.6, 1.0 Hz, 4H), 1.74-1.65 (sept, J=6.6 Hz, 1H), 1.65-1.55 (m, 4H), 1.55-1.37 (m, 9H), 1.34-1.12 (m, 33H), 0.93-0.82 (m, 12H); 13 C NMR (151 MHz, CDCl3) δ 174.1, 173.5, 74.4, 69.0, 58.5, 55.6, 53.9, 53.6, 34.6, 34.5, 34.3, 34.3, 32.0, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 27.5, 27.3, 26.8, 26.2, 25.5, 25.2, 23.1, 22.8, 16.5, 14.2, 11.4.

[0344] Pentan-3-yl 9-((2-hydroxyethyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)nonanoate (compound 74): [ka]

[0345] Compound 74 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (560 mg, 0.856 mmol, 82%). MS (ESI): m / z [M+H] + 655.3 for C 40 H 80 NO5; 1H NMR (600 MHz, CDCl3) δ 4.89-4.83 (m, 1H), 4.75 (tt, J=7.1, 5.3 Hz, 1H), 3.52 (t, J=5.4 Hz, 2H), 2.57 (t, J=5.4 Hz, 2H), 2.49-2.40 (m, 4H), 2.29 (td, J=7.5, 5.1 Hz, 4H), 1.65-1.39 (m, 16H), 1.34-1.20 (m, 36H), 0.87 (t, J=7.3 Hz, 12H); 13 C NMR (151 MHz, CDCl3) δ 173.9, 173.5, 76.5, 74.4, 58.5, 55.6, 53.9, 53.6, 34.8, 34.6, 34.3, 32.0, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.5, 27.3, 26.8, 26.6, 25.5, 25.3, 23.1, 22.8, 14.2, 9.7.

[0346] Pentan-3-yl 10-((2-hydroxyethyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 75): [ka]

[0347] Compound 75 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (510 mg, 0.763 mmol, 76%). MS (ESI): m / z [M+H] + 668.7 for C 41 H 82 NO5; 1H NMR (600 MHz, CDCl3) δ 4.89-4.83 (m, 1H), 4.75 (tt, J=7.1, 5.3 Hz, 1H), 3.52 (t, J=5.4 Hz, 2H), 2.88 (bs, 1H), 2.57 (t, J=5.4 Hz, 2H), 2.47 (t, J=7.8 Hz, 2H), 2.44 (t, J=7.8 Hz, 2H), 2.30 (t, J=7.5 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.65-1.39 (m, 16H), 1.33-1.19 (m, 38H), 0.87 (t, J=7.4 Hz, 12H); 13 C NMR (151 MHz, CDCl3) δ 173.9, 173.4, 76.5, 74.4, 58.5, 55.7, 54.0, 53.6, 34.8, 34.6, 34.3, 32.0, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.6, 27.3, 26.8, 26.6, 25.5, 25.3, 23.1, 22.8, 14.2, 9.7.

[0348] 3-Methylbut-2-en-1-yl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 76): [ka]

[0349] Compound 76 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (240 mg, 0.385 mmol, 55%). MS (ESI): m / z [M+H] + 624.5 for C 38 H 74 NO5; 1H NMR (600 MHz, CDCl3) δ 5.33 (tsept, J=7.2, 1.2 Hz, 1H), 4.89-4.83 (m, 1H), 4.56 (d, J=7.2 Hz, 2H), 3.56 (t, J=5.3 Hz, 2H), 3.07 (bs, 1H), 2.62 (t, J=5.4 Hz, 2H), 2.52 (t, J=7.5 Hz, 2H), 2.48 (t, J=7.5 Hz, 2H), 2.30 (t, J=7.2 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.77-1.74 (m, 3H), 1.70 (d, J=0.9 Hz, 3H), 1.65-1.58 (m, 4H), 1.55-1.42 (m, 8H), 1.33-1.20 (m, 32H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 174.0, 173.4, 139.0, 118.9, 74.5, 61.3, 58.3, 55.7, 54.0, 53.7, 34.5, 34.5, 34.3, 32.0, 29.7, 29.6, 29.5, 29.4, 29.4, 29.2, 27.5, 26.9, 26.5, 25.9, 25.5, 25.1, 23.1, 22.8, 18.1, 14.2.

[0350] 3-Methylbut-2-en-1-yl 9-((2-hydroxyethyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)nonanoate (compound 77): [ka]

[0351] Compound 77 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (240 mg, 0.368 mmol, 53%). MS (ESI): m / z [M+H] + 652.6 for C 40 H 78 NO5; 1H NMR (600 MHz, CDCl3) δ 5.33 (tsept, J=7.2, 1.2 Hz, 1H), 4.89-4.83 (m, 1H), 4.56 (d, J=7.2 Hz, 2H), 3.56 (t, J=5.3 Hz, 2H), 3.07 (bs, 1H), 2.62 (t, J=5.4 Hz, 2H), 2.52 (t, J=7.5 Hz, 2H), 2.48 (t, J=7.5 Hz, 2H), 2.30 (t, J=7.2 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 1.77-1.74 (m, 3H), 1.70 (d, J=0.9 Hz, 3H), 1.65-1.58 (m, 4H), 1.55-1.42 (m, 8H), 1.33-1.20 (m, 32H), 0.87 (t, J=6.9 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 174.0, 173.4, 139.0, 118.9, 74.5, 61.3, 58.3, 55.7, 54.0, 53.7, 34.5, 34.5, 34.3, 32.0, 29.7, 29.6, 29.5, 29.4, 29.4, 29.2, 27.5, 26.9, 26.5, 25.9, 25.5, 25.1, 23.1, 22.8, 18.1, 14.2.

[0352] Pent-3-yn-1-yl 9-((5-(heptadecan-9-yloxy)-5-oxopentyl)(2-hydroxyethyl)amino)nonanoate (compound 78): [ka]

[0353] Compound 78 was synthesized according to Representative Procedure 1 and General Methods A, B, C, and G. The product was obtained as a colorless oil (350 mg, 0.563 mmol, 80%). MS (ESI): m / z [M+H] + 623.1 for C 38 H 72 NO5; 1H NMR (600 MHz, CDCl3) δ 4.87 (qu, J=6.3 Hz, 1H), 4.13 (t, J=7.0 Hz, 2H), 3.56 (t, J=5.3 Hz, 2H), 3.13 (bs, 1H), 2.62 (t, J=5.3 Hz, 2H), 2.52 (t, J=7.5 Hz, 2H), 2.48 (t, J=7.3 Hz, 2H), 2.45 (m, J=2.6 Hz, 2H), 2.31 (t, J=7.4 Hz, 4H), 1.78 (t, J=2.5 Hz, 3H), 1.62 (qu, J=7.4 Hz, 4H), 1.54-1.48 (m, 6H), 1.45 (m, J=7.2 Hz, 2H), 1.30-1.26 (m, 32H), 0.88 (t, J=7.0 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 173.6, 173.2, 77.2, 74.7, 74.3, 62.6, 58.2, 55.5, 53.8, 53.5, 34.4, 34.2, 34.1, 31.8, 29.5, 29.5, 29.3, 29.2, 29.0, 27.3, 26.8, 26.3, 25.3, 24.9, 22.9, 22.6, 19.2, 14.1, 3.4.

[0354] Pentan-3-yl 10-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)decanoate (compound 79): [ka]

[0355] Compound 79 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (580 mg, 0.869 mmol, 93%). MS (ESI): m / z [M+H] + 667.7 for C 41 H 82 N2O4; 1H NMR (400 MHz, CDCl3) δ 4.85 (qu J=6.3 Hz, 1H), 4.74 (tt, J=7.0, 5.4 Hz, 1H), 2.68 (t, J=6.6 Hz, 2H), 2.42-2.31 (m, 6H), 2.28 (t, J=7.6 Hz, 2H), 2.28 (t, J=7.6 Hz, 2H), 1.66-1.11 (m, 56H), 0.86 (t, J=7.4 Hz, 12H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.6, 76.5, 74.3, 54.3, 54.2, 53.8, 42.4, 34.8, 34.8, 34.3, 32.1, 32.0, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.7, 27.3, 26.8, 26.6, 25.5, 25.3, 24.7, 23.3, 22.8, 14.2, 9.7.

[0356] Pentan-3-yl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)amino)decanoate (compound 80): [ka]

[0357] Compound 80 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (310 mg, 0.399 mmol, 89%). MS (ESI): m / z [M+H] + 777.3 for C 46 H 86 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.61 (bs, 1H), 7.37 (bs, 1H), 4.83 (qu, J=6.3 Hz, 1H), 4.73 (tt, J=7.1, 5.3 Hz, 1H), 3.65 (bs, 2H), 3.37-3.27 (m, 3H), 2.42-2.32 (m, 6H), 2.31-2.25 (m, 4H), 1.66-1.33 (m, 20H), 1.33-1.17 (m, 34H), 0.88-0.84 (m, 12H); 13 C NMR (151 MHz, CDCl3) δ 183.0, 182.4, 173.9, 173.9, 168.4, 168.2, 76.5, 74.5, 54.2, 53.7, 44.6, 34.8, 34.7, 34.2, 32.0, 31.4, 29.7, 29.7, 29.6, 29.6, 29.4, 29.4, 29.4, 29.3, 29.2, 27.8, 27.1, 26.6, 26.6, 25.5, 25.3, 24.3, 23.3, 22.8, 14.2, 9.7.

[0358] Pentan-3-yl 10-((4-aminobutyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 81): [ka]

[0359] Compound 81 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (570 mg, 0.820 mmol, 91%). MS (ESI): m / z [M+H] + 695.7 for C 43 H 87 N2O4; 1H NMR (400 MHz, CDCl3) δ 4.85 (qu J=6.2 Hz, 1H), 4.74 (tt, J=7.0, 5.5 Hz, 1H), 2.68 (t, J=6.6 Hz, 2H), 2.43-2.31 (m, 6H), 2.28 (t, J=7.6 Hz, 2H), 2.28 (t, J=7.4 Hz, 2H), 1.66-1.15 (m, 60H), 0.86 (t, J=7.4 Hz, 12H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.6, 76.5, 74.3, 54.3, 54.2, 53.8, 42.4, 34.8, 34.8, 34.3, 32.0, 32.0, 29.7, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.7, 27.3, 26.8, 26.6, 25.5, 25.3, 24.7, 23.3, 22.8, 14.2, 9.7.

[0360] Pentan-3-yl 10-((4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 82): [ka]

[0361] Compound 82 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (330 mg, 0.410 mmol, 95%). MS (ESI): m / z [M+H] + 805.4 for C 48 H 90 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.61 (bs, 1H), 7.36 (bs, 1H), 4.83 (qu, J=6.3 Hz, 1H), 4.73 (tt, J=7.1, 5.3 Hz, 1H), 3.65 (bs, 2H), 3.31 (d, J=5.1 Hz, 3H), 2.42-2.32 (m, 6H), 2.31-2.25 (m, 4H), 1.67-1.33 (m, 20H), 1.32-1.17 (m, 38H), 0.86 (t, J=7.3 Hz, 12H); 13 C NMR (151 MHz, CDCl3) δ 183.0, 182.4, 173.9, 173.9, 168.4, 168.2, 76.5, 74.5, 54.2, 53.7, 44.6, 34.8, 34.7, 34.2, 32.0, 31.4, 29.7, 29.7, 29.7, 29.7, 29.4, 29.4, 29.4, 29.3, 29.2, 27.8, 27.1, 26.7, 26.6, 25.5, 25.3, 24.3, 23.3, 22.8, 14.2, 9.7.

[0362] Pentan-3-yl 10-((3-aminopropyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)decanoate (compound 83): [ka]

[0363] Compound 83 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (540 mg, 0.827 mmol, 94%). MS (ESI): m / z [M+H] + 653.7 for C 40 H 81 N2O4; 1H NMR (400 MHz, CDCl3) δ 4.90-4.80 (qu J=4.6 Hz, 1H), 4.74 (tt, J=7.0, 5.4 Hz, 1H), 2.70 (t, J=7.0 Hz, 2H), 2.46-2.24 (m, 6H), 2.28 (t, J=7.4 Hz, 2H), 2.28 (t, J=7.6 Hz, 2H), 1.66-1.17 (m, 54H), 0.86 (t, J=7.4 Hz, 12H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.6, 76.5, 74.3, 54.3, 53.9, 52.0, 41.0, 34.8, 34.7, 34.3, 32.0, 31.2, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.7, 27.2, 26.7, 26.6, 25.5, 25.3, 23.3, 22.8, 14.2, 9.7.

[0364] Pentan-3-yl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)decanoate (compound 84): [ka]

[0365] Compound 84 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a yellowish, waxy amorphous solid (320 mg, 0.420 mmol, 91%). MS (ESI): m / z [M+H] + 763.1 for C 45 H 84 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.38 (bs, 1H), 7.00 (bs, 1H), 4.81 (qu J=6.3 Hz, 1H), 4.73 (tt, J=7.1, 5.3 Hz, 1H), 3.69 (bs, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.48 (t, J=6.4 Hz, 2H), 2.40 (t, J=7.0 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.31 (t, J=7.2 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H), 1.75 (qu J=6.4 Hz, 2H), 1.64-1.47 (m, 12H), 1.47-1.34 (m, 4H), 1.33-1.17 (m, 34H), 0.86 (t, J=7.4 Hz, 12H); 13 C NMR (151 MHz, CDCl3) δ 183.3, 183.0, 174.3, 173.9, 168.5, 168.3, 76.5, 74.8, 53.9, 53.6, 51.3, 43.4, 34.8, 34.5, 34.2, 32.0, 31.3, 29.7, 29.6, 29.6, 29.6, 29.4, 29.3, 29.3, 28.2, 27.7, 26.7, 26.6, 26.4, 25.4, 25.3, 23.1, 22.8, 14.2, 9.7.

[0366] Pentan-3-yl 10-((3-aminopropyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 85): [ka]

[0367] Compound 85 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (540 mg, 0.793 mmol, 97%). MS (ESI): m / z [M+H] + 681.7 for C 42 H 85 N2O4;1 H NMR (400 MHz, CDCl3) δ 4.84 (qu J=6.3 Hz, 1H), 4.74 (tt, J=7.0, 5.4 Hz, 1H), 2.70 (t, J=6.8 Hz, 2H), 2.46-2.31 (m, 6H), 2.28 (t, J=7.6 Hz, 2H), 2.27 (t, J=7.6 Hz, 2H), 1.66-1.14 (m, 58H), 0.86 (t, J=7.4 Hz, 12H); 13 C NMR (101 MHz, CDCl3) δ 173.9, 173.6, 76.5, 74.3, 54.3, 53.9, 52.0, 41.0, 34.8, 34.7, 34.3, 32.0, 31.2, 29.7, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 27.7, 27.2, 26.7, 26.6, 25.5, 25.3, 23.3, 22.8, 14.2, 9.7.

[0368] Pentan-3-yl 10-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 86): [ka]

[0369] Compound 86 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (320 mg, 0.405 mmol, 92%). MS (ESI): m / z [M+H] + 791.1 for C 47 H 88 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.32 (bs, 1H), 6.92 (bs, 1H), 4.81 (qu, J=6.2 Hz, 1H), 4.73 (tt, J=7.1, 5.3 Hz, 1H), 3.70 (bs, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.48 (t, J=6.4 Hz, 2H), 2.40 (t, J=7.0 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.32 (t, J=7.2 Hz, 2H), 2.28 (t, J=7.5 Hz, 2H), 1.75 (qu, J=6.3 Hz, 2H), 1.63-1.47 (m, 12H), 1.47-1.34 (m, 4H), 1.33-1.17 (m, 38H), 0.88-0.82 (m, 12H); 13 C NMR (151 MHz, CDCl3) δ 183.3, 183.0, 174.3, 173.9, 168.5, 168.3, 76.5, 74.9, 53.9, 53.6, 51.3, 43.4, 34.8, 34.5, 34.2, 32.0, 31.3, 29.7, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 28.1, 27.7, 26.7, 26.6, 26.5, 25.5, 25.3, 23.1, 22.8, 14.2, 9.7.

[0370] Pentyl 10-((4-aminobutyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)decanoate (compound 87): [ka]

[0371] Compound 87 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (560 mg, 0.839 mmol, 90%). MS (ESI): m / z [M+H] + 667.7 for C 41 H 83 N2O4; 1H NMR (400 MHz, CDCl3) δ 4.84 (qu J=6.3 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 2.68 (t, J=6.6 Hz, 2H), 2.43-2.32 (m, 6H), 2.28 (t, J=7.6 Hz, 2H), 2.27 (t, J=7.4 Hz, 2H), 1.67-1.16 (m, 58H), 0.94-0.81 (m, 9H); 13 C NMR (101 MHz, CDCl3) δ 174.1, 173.6, 74.3, 64.5, 54.3, 54.2, 53.8, 42.4, 34.8, 34.5, 34.3, 32.1, 32.0, 29.7, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 27.7, 27.2, 26.7, 25.5, 25.2, 24.7, 23.3, 22.8, 22.4, 14.2, 14.1.

[0372] Pentyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)amino)decanoate (compound 88): [ka]

[0373] Compound 88 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (260 mg, 0.335 mmol, 75%). MS (ESI): m / z [M+H] + 777.2 for C 46 H 86 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.62 (bs, 1H), 7.39 (bs, 1H), 4.83 (qu J=6.2 Hz, 1H), 4.03 (t, J=6.8 Hz, 2H), 3.65 (d, J=5.0 Hz, 2H), 3.31 (d, J=5.0 Hz, 3H), 2.45-2.31 (m, 6H), 2.31-2.23 (m, 4H), 1.68-1.54 (m, 8H), 1.53-1.15 (m, 48H), 0.89 (t, J=7.2 Hz, 3H), 0.86 (t, J=7.0 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 183.0, 182.3, 174.1, 173.9, 168.4, 168.2, 74.5, 64.5, 54.2, 53.7, 44.6, 34.7, 34.5, 34.2, 32.0, 31.4, 31.2, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 29.2, 28.5, 28.2, 27.8, 27.1, 26.6, 25.4, 25.1, 24.2, 23.3, 22.8, 22.4, 14.2, 14.1.

[0374] Pentyl 10-((3-aminopropyl)(5-(heptadecan-9-yloxy)-5-oxopentyl)amino)decanoate (compound 89): [ka]

[0375] Compound 89 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (500 mg, 0.766 mmol, 92%). MS (ESI): m / z [M+H] + 653.7 for C 40 H 81 N2O4; 1H NMR (600 MHz, CDCl3) δ 4.85 (qu J=6.3 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 2.70 (t, J=6.8 Hz, 2H), 2.42 (t, J=6.9 Hz, 2H), 2.38 (t, J=7.5 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.28 (t, J=7.2 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H), 1.65-1.10 (m, 56H), 0.89 (t, J=7.2 Hz, 3H), 0.86 (t, J=7.2 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 174.1, 173.6, 74.3, 64.5, 54.3, 53.9, 52.0, 41.0, 34.7, 34.5, 34.3, 32.0, 31.2, 29.7, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 27.7, 27.2, 26.7, 25.4, 25.1, 23.3, 22.8, 22.4, 14.2, 14.1.

[0376] Pentyl 10-((5-(heptadecan-9-yloxy)-5-oxopentyl)(3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)amino)decanoate (compound 90): [ka]

[0377] Compound 90 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (210 mg, 0.276 mmol, 60%). MS (ESI): m / z [M+H] + 763.3 for C 45 H 84 N3O6; 1H NMR (400 MHz, CDCl3) δ 7.45 (bs, 1H), 7.12 (bs, 1H), 4.81 (qu, J=6.3 Hz, 1H), 4.03 (t, J=6.8 Hz, 2H), 3.68 (bs, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.47 (t, J=6.5 Hz, 2H), 2.43-2.23 (m, 8H), 1.75 (qu, J=6.4 Hz, 2H), 1.66-1.13 (m, 52H), 0.92-0.79 (m, 9H); 13 C NMR (101 MHz, CDCl3) δ 183.2, 182.8, 174.2, 174.1, 168.4, 168.3, 74.7, 64.5, 53.9, 53.6, 51.3, 43.3, 34.5, 34.5, 34.2, 31.9, 31.3, 29.7, 29.6, 29.6, 29.6, 29.4, 29.3, 29.3, 28.4, 28.3, 28.2, 27.7, 26.7, 26.5, 25.4, 25.1, 23.1, 22.8, 22.4, 14.2, 14.1.

[0378] Pentyl 9-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)nonanoate (compound 91): [ka]

[0379] Compound 91 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (810 mg, 1.040 mmol, 80%). MS (ESI): m / z [M+H] + 777.2 for C 46 H 86 N3O6; 1H NMR (600 MHz, CDCl3) δ 7.19 (bs, 1H), 6.74 (bs, 1H), 4.82 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.72 (bs, 2H), 3.30 (d, J=5.0 Hz, 3H), 2.50 (bt, J=6.1 Hz, 2H), 2.42 (bt, J=6.9 Hz, 2H), 2.37 (bt, J=6.5 Hz, 2H), 2.34 (t, J=7.2 Hz, 2H), 2.29 (t, J=7.5 Hz, 2H), 2.01 (bs, 1H), 1.76 (qu, J=6.2 Hz, 2H), 1.62 (sext, J=7.1 Hz, 6H), 1.53-1.52 (m, 4H), 1.48-1.37 (m, 4H), 1.36-1.30 (m, 4H), 1.29-1.25 (m, 36H), 0.91 (t, J=7.1 Hz, 3H), 0.88 (t, J=7.1 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 183.2, 183.1, 174.4, 174.0, 168.4, 168.1, 74.9, 64.4, 53.6, 53.4, 51.1, 43.2, 34.3, 34.3, 34.0, 31.8, 31.2, 29.5, 29.5, 29.4, 29.3, 29.1, 28.3, 28.0, 27.8, 27.5, 26.3, 25.3, 24.9, 22.9, 22.6, 22.3, 14.1, 13.9.

[0380] Pentyl 9-((4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)nonanoate (compound 92): [ka]

[0381] Compound 92 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (1.17 g, 1.480 mmol, 80%). MS (ESI): m / z [M+H] + 791.2 for C 48 H 87 N3O6; 1 H NMR (400 MHz, CDCl3) δ 7.46 (bs, 1H), 7.21 (bs, 1H), 4.84 (qu, J=6.2 Hz, 1H), 4.05 (t, J=6.8 Hz, 2H), 3.66 (bd, J=4.9 Hz, 2H), 3.33 (d, J=5.0 Hz, 3H), 2.42-2.36 (m, 6H), 2.34-2,26 (m, 4H), 1.67-1.59 (m, 8H), 1.52-1.45 (m, 8H), 1.39-1.25 (m, 42H), 0.91 (t, J=6.9 Hz, 3H), 0.87 (t, J=6.8 Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 182.9, 182.4, 174.0, 173.9, 168.2, 168.2, 74.5, 64.4, 54.0, 53.5, 44.3, 34.5, 34.3, 34.1, 31.9, 31.2, 29.5, 29.4, 29.3, 29.1, 29.0, 28.3, 28.1, 27.5, 26.8, 26.3, 25.3, 25.0, 24.0, 23.1, 22.6, 22.3, 14.1, 13.9.

[0382] Pentyl 10-((3-aminopropyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 93): [ka]

[0383] Compound 93 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (510 mg, 0.749 mmol, 93%). MS (ESI): m / z [M+H] + 681.7 for C 42 H 85 N2O4; 1 H NMR (400 MHz, CDCl3) δ 4.85 (qu J=6.2 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 2.70 (t, J=6.6 Hz, 2H), 2.42 (t, J=7.2 Hz, 2H), 2.38 (t, J=7.5 Hz, 2H), 2.35 (t, J=7.5 Hz, 2H), 2.28 (t, J=7.5 Hz, 2H), 2.27 (t, J=7.8 Hz, 2H), 1.66-1.07 (m, 60H), 0.89 (t, J=7.2 Hz, 3H), 0.86 (t, J=7.2 Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 174.1, 173.6, 74.3, 64.5, 54.3, 53.9, 52.0, 41.0, 34.7, 34.5, 34.3, 32.0, 31.2, 29.7, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 27.7, 27.2, 26.7, 25.5, 25.1, 23.3, 22.8, 22.4, 14.2, 14.1.

[0384] Pentyl 10-((3-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)propyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 94): [ka]

[0385] Compound 94 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (260 mg, 0.329 mmol, 75%). MS (ESI): m / z [M+H] + 791.4 for C 47 H 88 N3O6; 1 H NMR (400 MHz, CDCl3) δ 7.34 (bs, 1H), 6.98 (bs, 1H), 4.81 (qu J=6.2 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 3.69 (bs, 2H), 3.29 (d, J=5.0 Hz, 3H), 2.48 (t, J=6.4 Hz, 2H), 2.40 (t, J=7.0 Hz, 2H), 2.35 (t, J=7.4 Hz, 2H), 2.32 (t, J=7.2 Hz, 2H), 2.27 (t, J=7.6 Hz, 2H), 1.75 (qu J=6.3 Hz, 2H), 1.66-1.14 (m, 56H), 0.89 (t, J=6.8 Hz, 3H), 0.86 (t, J=7.0 Hz, 6H); 13 C NMR (101 MHz, CDCl3) δ 183.3, 183.0, 174.3, 174.1, 168.5, 168.3, 74.9, 64.5, 53.9, 53.6, 51.3, 43.3, 34.5, 34.2, 32.0, 31.3, 29.7, 29.7, 29.6, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 28.2, 27.7, 26.7, 26.4, 25.4, 25.1, 23.1, 22.8, 22.4, 14.2, 14.1.

[0386] Pentyl 10-((4-aminobutyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 95): [ka]

[0387] Compound 95 was synthesized according to Representative Procedure 2 and General Methods A, C, D, E, and H. The product was obtained as a colorless oil (600 mg, 0.863 mmol, 95%). MS (ESI): m / z [M+H] + 695.7 for C 43 H 87 N2O4; 1 H NMR (400 MHz, CDCl3) δ 4.84 (qu J=6.3 Hz, 1H), 4.04 (t, J=6.8 Hz, 2H), 2.68 (t, J=6.6 Hz, 2H), 2.44-2.31 (m, 6H), 2.28 (t, J=7.6 Hz, 2H), 2.27 (t, J=7.6 Hz, 2H), 1.66-1.14 (m, 60H), 0.92-0.82 (m, 9H); 13 C NMR (101 MHz, CDCl3) δ 174.1, 173.6, 74.3, 64.5, 54.2, 54.1, 53.8, 42.3, 34.7, 34.5, 34.3, 32.0, 31.9, 29.7, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 28.5, 28.2, 27.7, 27.2, 26.7, 25.4, 25.1, 24.7, 23.3, 22.8, 22.4, 14.2, 14.1.

[0388] Pentyl 10-((4-((2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl)amino)butyl)(5-(nonadecan-10-yloxy)-5-oxopentyl)amino)decanoate (compound 96): [ka]

[0389] Compound 96 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a colorless, waxy amorphous solid (240 mg, 0.299 mmol, 69%). MS (ESI): m / z [M+H] +805.4 for C 48 H 90 N3O6; 1 H NMR (600 MHz, CDCl3) δ 7.54 (bs, 1H), 7.29 (bs, 1H), 4.83 (qu J=6.3 Hz, 1H), 4.04 (t, J=6.6 Hz, 2H), 3.65 (bs, 2H), 3.31 (d, J=5.1 Hz, 3H), 2.45-2.32 (m, 6H), 2.29 (t, J=7.5 Hz, 2H), 2.27 (t, J=7.5 Hz, 2H), 1.68-1.55 (m, 8H), 1.54-1.11 (m, 52H), 0.89 (t, J=7.2 Hz, 3H), 0.86 (t, J=7.2 Hz, 6H); 13 C NMR (151 MHz, CDCl3) δ 183.0, 182.5, 174.1, 174.0, 168.4, 168.3, 74.6, 64.5, 54.2, 53.7, 44.6, 34.7, 34.5, 34.2, 32.0, 31.4, 29.7, 29.7, 29.7, 29.6, 29.4, 29.4, 29.3, 29.2, 28.5, 28.2, 27.8, 25.5, 25.1, 24.2, 23.3, 22.8, 22.4, 14.2, 14.1.

[0390] 9-[(2-hydroxyethyl)[5-(nonadecan-10-yloxy)-5-oxopentyl]amino]nonyl hexanoate (compound 97): [ka]

[0391] Compound 97 was synthesized according to Representative Procedure 1 using Intermediate H and general methods F, B, C, and G. The product was obtained as a pale yellow oil (240 mg, 0.498 mmol, 71%). MS (ESI): m / z [M+H] + 669.3 for C 41 H 81 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.90-4.85 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.60 (t,J=5.9 Hz, 2H), 2.63 (t,J=5.9 Hz, 2H), 2.54-2.50 (m, 2H), 2.49-2.47 (m, 2H), 2.34 (t,J=7.4 Hz, 2H), 2.31 (t,J=7.5 Hz, 2H), 1.66-1.61 (m, 6H), 1.54-1.50 (m, 6H), 1.49-1.44 (m, 2H), 1.38-1.27 (m, 43H), 0.91 (t,J=7.1 Hz, 3H), 0.89 (t,J=7.0 Hz, 6H); 13 C NMR (151 MHz, CDCl3-CD3OD=2:1 v / v)δ175.1, 174.4, 75.1, 65.0, 59.2, 56.1, 54.6, 54.1, 34.8, 34.7, 34.4, 32.2, 31.6, 29.9, 29.9, 29.8, 29.8, 29.6, 29.5, 28.9, 27.8, 27.0, 26.4, 26.3, 25.7, 25.0, 23.4, 23.0, 22.6, 14.2, 14.0.

[0392] 11-{[5-(heptadecan-9-yloxy)-5-oxopentyl](2-hydroxyethyl)amino}undecylhexanoate (compound 98): [ka]

[0393] Compound 98 was synthesized according to Representative Procedure 1 using Intermediate H and general methods F, B, C, and G. The product was obtained as a pale yellow oil (260 mg, 0.455 mmol, 84%). MS (ESI): m / z [M+H] + 669.1 for C 41 H 81 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.90-4.85 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.61 (t,J=5.9 Hz, 2H), 2.64 (t,J=5.9 Hz, 2H), 2.55-2.53 (m, 2H), 2.52-2.49 (m, 2H), 2.35 (t,J=7.4 Hz, 2H), 2.31 (t,J=7.5 Hz, 2H), 1.66-1.61 (m, 6H), 1.55-1.51 (m, 6H), 1.49-1.44 (m, 2H), 1.37-1.28 (m, 43H), 0.91 (t,J=7.1 Hz, 3H), 0.89 (t,J=7.0 Hz, 6H); 13 C NMR (151 MHz, CDCl3-CD3OD=2:1 v / v)δ175.1, 174.4, 75.1, 65.0, 59.1, 56.1, 54.5, 54.1, 34.8, 34.7, 34.4, 32.2, 31.6, 29.9, 29.9, 29.9, 29.8, 29.8, 29.6, 28.9, 27.9, 26.9, 26.3, 26.3, 25.7, 25.0, 23.3, 23.0, 22.6, 14.2, 14.0.

[0394] 11-[(2-hydroxyethyl)[5-(nonadecan-10-yloxy)-5-oxopentyl]amino]undecylhexanoate (compound 99): [ka]

[0395] Compound 99 was synthesized according to Representative Procedure 1 using Intermediate H and general methods F, B, C, and G. The product was obtained as a pale yellow oil (230 mg, 0.455 mmol, 72%). MS (ESI): m / z [M+H] + 697.4 for C 43 H 85 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.90-4.85 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.60 (t,J=5.9 Hz, 2H), 2.63 (t,J=5.9 Hz, 2H), 2.54-2.52 (m, 2H), 2.50-2.48 (m, 2H), 2.34 (t,J=7.4 Hz, 2H), 2.31 (t,J=7.5 Hz, 2H), 1.66-1.61 (m, 6H), 1.54-1.50 (m, 6H), 1.49-1.44 (m, 2H), 1.37-1.27 (m, 47H), 0.91 (t,J=7.1 Hz, 3H), 0.89 (t,J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1 v / v)δ175.1, 174.3, 75.1, 65.0, 59.2, 56.1, 54.5, 54.1, 34.8, 34.7, 34.4, 32.2, 31.6, 29.9, 29.9, 29.9, 29.8, 29.6, 29.6, 28.9, 27.9, 26.9, 26.4, 26.3, 25.7, 25.0, 23.4, 23.0, 22.6, 14.2, 14.0.

[0396] 9-[(2-hydroxyethyl)[5-(nonadecan-10-yloxy)-5-oxopentyl]amino]nonyl hexanoate (compound 100): [ka]

[0397] Compound 100 was synthesized according to Representative Procedure 1 using Intermediate H and general methods F, B, C, and G. The product was obtained as a pale yellow oil (240 mg, 0.356 mmol, 71%). MS (ESI): m / z [M+H] + 669.3 for C 41 H 82 NO5; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.90-4.85 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.60 (t,J=5.9 Hz, 2H), 2.63 (t,J=5.9 Hz, 2H), 2.54-2.50 (m, 2H), 2.49-2.47 (m, 2H), 2.34 (t,J=7.4 Hz, 2H), 2.31 (t,J=7.5 Hz, 2H), 1.66-1.61 (m, 6H), 1.54-1.50 (m, 6H), 1.49-1.44 (m, 2H), 1.38-1.27 (m, 43H), 0.91 (t,J=7.1 Hz, 3H), 0.89 (t,J=7.0 Hz, 6H); 13 C NMR (151 MHz, CDCl3-CD3OD=2:1 v / v)δ175.1, 174.4, 75.1, 65.0, 59.2, 56.1, 54.6, 54.1, 34.8, 34.7, 34.4, 32.2, 31.6, 29.9, 29.9, 29.8, 29.8, 29.6, 29.5, 28.9, 27.8, 27.0, 26.4, 26.3, 25.7, 25.0, 23.4, 23.0, 22.6, 14.2, 14.0.

[0398] 5-{[5-(heptadecan-9-yloxy)-5-oxopentyl](3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)amino}pentyl hexanoate (compound 101): [ka]

[0399] Compound 101 was synthesized according to Representative Procedures 2 and 3 and General Methods A, C, D, E, G, and I. The product was obtained as a pale yellow oil (50 mg, 0.070 mmol, 55%). MS (ESI): m / z [M+H] + 706.1 for C 41 H 76 N3O6; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.84-4.80 (m, 1H), 4.04 (t, J=6.5 Hz, 2H), 3.65-3.62 (m, 2H), 3.25 (bs, 3H), 2.96-2.87 (m, 5H), 2.34 (t, J=6.1 Hz, 2H), 2.27 (t, J=7.6 Hz, 2H), 1.93 (bs, 2H), 1.67-1.54 (m, 9H), 1.49-1.48 (m, 4H), 1.39-1.34 (m, 2H), 1.31-1.23 (m, 30H), 0.86-0.83 (m, 9H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1 v / v)δ182.4, 182.0, 174.2, 173.1, 168.7, 167.5, 74.7, 63.6, 52.8, 52.6, 50.5, 41.2, 33.9, 33.7, 33.5, 31.5, 31.0, 30.9, 30.5, 29.2, 29.2, 28.9, 28.0, 26.2, 25.0, 24.3, 23.8, 23.6, 23.1, 22.3, 22.0, 22.0, 13.6, 13.4.

[0400] 9-[(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)[5-(nonadecan-10-yloxy)-5-oxopentyl]amino]nonyl hexanoate (compound 102): [ka]

[0401] Compound 102 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (520 mg, 0.858 mmol, 61%). MS (ESI): m / z [M+H] + 791.4 for C 47 H 87 N3O6; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.90-4.84 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.63 (bs, 2H), 3.28 (bs, 3H), 2.54-2.42 (m, 6H), 2.37-2.29 (m, 4H), 1.80-1.73 (m, 2H), 1.67-1.59 (m, 6H), 1.56-1.41 (m, 8H), 1.39-1.25 (m, 45H), 0.93-0.87 (m, 9H); 13 C NMR (151 MHz, CDCl3-CD3OD=2:1 v / v)δ182.6, 182.4, 174.7, 173.2, 169.4, 167.6, 75.1, 64.5, 53.0, 52.6, 50.7, 40.8, 34.3, 34.0, 33.5, 31.8, 31.2, 30.7, 29.5, 29.5, 29.4, 29.2, 29.2, 29.0, 28.5, 26.6, 25.8, 25.3, 24.6, 23.6, 23.1, 22.6, 22.2, 21.9, 13.8, 13.6.

[0402] 11-[(3-{[2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl]amino}propyl)[5-(nonadecan-10-yloxy)-5-oxopentyl]amino]undecylhexanoate (compound 103): [ka]

[0403] Compound 103 was synthesized according to Representative Procedures 2 and 3, and General Methods A, C, D, E, H, and I. The product was obtained as a pale yellow oil (580 mg, 0.748 mmol, 77%). MS (ESI): m / z [M+H] + 819.2 for C 49 H 91 N3O6; 1H NMR (600 MHz, CDCl3-CD3OD=2:1 v / v) δ 4.88-4.84 (m, 1H), 4.07 (t,J=6.7 Hz, 2H), 3.63 (m, 2H), 3.28 (bs, 3H), 2.52-2.42 (m, 6H), 2.35 (t,J=7.4 Hz, 2H), 2.31 (t,J=7.5 Hz, 2H), 1.78-1.73 (m, 2H), 1.66-1.59 (m, 6H), 1.55-1.48 (m, 6H), 1.47-1.41 (m, 2H), 1.37-1.27 (m, 49H), 0.91 (t,J=7.1 Hz, 3H), 0.88 (t,J=7.1 Hz, 6H); 13 C NMR (150 MHz, CDCl3-CD3OD=2:1 v / v)δ182.9, 182.7, 175.2, 173.6, 169.7, 167.9, 75.5, 64.9, 53.3, 52.9, 51.0, 41.2, 34.7, 34.3, 33.8, 32.2, 31.6, 31.1, 29.9, 29.9, 29.8, 29.8, 29.7, 29.6, 29.5, 29.5, 28.9, 27.0, 26.2, 26.2, 25.7, 25.0, 24.0, 23.5, 23.0, 22.6, 22.2, 14.2, 14.0.

[0404] The compounds listed above correspond to exemplary structures of Formula I shown in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20 Table 1-21 Table 1-22 Table 1-23 [Table 1-24] [Table 1-25] [Table 1-26] [Table 1-27] [Table 1-28]

[0405] Furthermore, it should be understood that any specific aspect of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Such aspects may be deemed to be known to those of skill in the art and therefore may be excluded even if the exclusion is not expressly set forth herein.

[0406] Although the present disclosure has been described in conjunction with its detailed description, it should be understood that the above description is intended to be illustrative, but not limiting, of the scope of the disclosure, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. Compounds of Formula I: 【Chemistry 1】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, or C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; Each R 6 are independently H, 【Chemistry 2】 or 【Transformation 3】 Selected from; Each M 1 and M 2 are independently selected from —C(O)O— and —OC(O)—, where M 1 or M 2 at least one of is -C(O)O-; each Q is selected from —O— or —NH—; and each G is -(CR 4 R 5 ) k - and; where each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

2. 10. The compound of claim 1, wherein the compound is: 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】 【Chemistry 4-4】 [Chemistry 4-5] [Chemistry 4-6] 【Chemistry 4-7】 【Chemistry 4-8】 【Chemistry 4-9】 【Chemistry 4-10】 【Chemistry 4-11】 【Chemistry 4-12】 【Chemistry 4-13】 【Chemistry 4-14】 【Chemistry 4-15】 【Chemistry 4-16】 【Chemistry 4-17】 【Chemistry 4-18】 【Chemistry 4-19】 【Chemistry 4-20】 【Chemistry 4-21】 【Chemistry 4-22】 【Chemistry 4-23】 【Chemistry 4-24】 【Chemistry 4-25】 or a salt or isomer thereof, The compound.

3. 10. The compound of claim 1, wherein the compound is 【Transformation 5】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

4. 10. The compound of claim 1, wherein the compound is 【Transformation 6】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; Each R 6 is H; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

5. 10. The compound of claim 1, wherein the compound is 【Transformation 7】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; Each R 6 teeth, 【Transformation 8】 and; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

6. 10. The compound of claim 1, wherein the compound is 【Chemistry 9】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; Each R 6 teeth, 【Chemistry 10】 and; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

7. 10. The compound of claim 1, wherein the compound is 【Chemistry 11】 or a salt or isomer thereof, wherein: each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The compound.

8. A nanoparticle composition comprising an ionizable lipid component comprising a compound according to any one of claims 1 to 7.

9. 10. The nanoparticle composition of claim 8, further comprising a bioactive agent.

10. The nanoparticle composition of any one of claims 8 to 9, further comprising one or more phospholipids.

11. 11. The nanoparticle composition of claim 10, 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-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME16.0). 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.

12. The nanoparticle composition of any one of claims 8 to 11, further comprising one or more structured lipids.

13. 13. The nanoparticle composition of claim 12, wherein the structural lipid comprises at least one of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, and α-tocopherol, and mixtures thereof.

14. The nanoparticle composition of any one of claims 8 to 13, further comprising one or more PEG lipids.

15. 15. The nanoparticle composition of claim 14, wherein the PEG comprises at least one of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

16. 16. The nanoparticle composition of any one of claims 8 to 15, further comprising a second ionizable lipid component other than the ionizable lipid component.

17. A pharmaceutical composition comprising the nanoparticle composition of any one of claims 8 to 16 and a pharmaceutically acceptable carrier.

18. A method for delivering a bioactive agent to a cell, the method comprising administering to a subject a nanoparticle composition described in any one of claims 8 to 16, wherein said administering comprises contacting a cell with the nanoparticle composition, thereby delivering the bioactive agent to the cell.

19. 20. A method for delivering a bioactive agent to a cell, the method comprising administering to a subject the pharmaceutical composition of claim 17.

20. 17. A method for producing a polypeptide of interest in a cell, the method comprising contacting the cell with the nanoparticle composition of any one of claims 8 to 16, wherein the bioactive agent is an mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest.

21. 17. A method for regulating expression of an endogenous nucleic acid in a cell, the method comprising contacting the cell with the nanoparticle composition of any one of claims 8 to 16, wherein the bioactive agent is an siRNA capable of binding to the endogenous nucleic acid, thereby enabling the siRNA to regulate expression of the endogenous nucleic acid.

22. The method of claim 21 , wherein the siRNA has a sequence complementary to a portion of an endogenous nucleic acid.

23. A method for regulating the expression of an endogenous nucleic acid in a cell, the method comprising contacting the cell with the nanoparticle composition of any one of claims 8 to 16, wherein the bioactive agent is an antisense RNA capable of binding to the endogenous nucleic acid, thereby enabling the siRNA to regulate the expression of the endogenous nucleic acid.

24. The method of claim 23, wherein the antisense RNA has a sequence complementary to a portion of the endogenous nucleic acid.

25. 17. A method of treating a subject having a disease or disorder, comprising administering to the subject a therapeutically effective amount of the nanoparticle composition of any one of claims 8 to 16.

26. A method for synthesizing the following compound: 【Chemistry 12】 The method comprising carrying out the following reaction: 【Chemistry 13】

27. A method for synthesizing the following compound: 【Chemistry 14】 The method comprising carrying out the following reaction: 【Chemistry 15】

28. A method for synthesizing the following compound: 【Chemistry 16】 The method comprising carrying out the following reaction: 【Chemistry 17】

29. A method for synthesizing the following compound: [Chemistry 18] (I)(a), the method comprising reacting: 【Chemistry 19】

30. A method for synthesizing the following compound: 【Chemistry 20】 The method comprising carrying out the following reaction: 【Chemistry 21】

31. 10. A method for synthesizing a compound of claim 3, comprising carrying out the following reaction: 【Chemistry 22】 During the ceremony each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The method.

32. 10. A method for synthesizing a compound of claim 4, comprising carrying out the following reaction: 【Chemistry 23】 During the ceremony each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; Each R 6 is H; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The method.

33. 10. A method for synthesizing a compound of claim 5, comprising carrying out the following reaction: 【Chemistry 24】 During the ceremony each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; R 6 teeth, 【Chemistry 25】 and; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The method.

34. 10. A method for synthesizing a compound of claim 6, comprising carrying out the following reaction: 【Chemistry 26】 During the ceremony each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is independently selected from C1-C5 alkyl, C2-C5 alkenyl, C2-C5 alkynyl, where the alkyl, alkenyl, or alkynyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; R 6 teeth, 【Chemistry 27】 and; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; And each R 4 and R 5 are independently H and C 1 ~C 3 selected from alkyl, The method.

35. 10. A method for synthesizing a compound of claim 7, comprising carrying out the following reaction: 【Chemistry 28】 During the ceremony X is Cl, Br; each m is independently an integer from 4 to 13; each n is independently an integer from 1 to 3; Each R 1 is R 1 is C 1 ~C 5 alkyl, where alkyl is linear or branched; Each R 2 and R 3 independently, C 1 ~C 14 alkyl; and each G is -(CR 4 R 5 ) k - and; wherein each k is selected from an integer from 2 to 5; The method.

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