Lipid compounds, compositions and their uses

Abstract

This disclosure provides lipid compounds and compositions comprising the lipid compounds of this disclosure (e.g., lipid nanoparticle (LNP) compositions). This disclosure provides a method for delivering an active agent (e.g., polynucleotide) to target cells or tissues, preferably extrahepatic cells or tissues, comprising administering an effective amount of the lipid nanoparticles of this disclosure to the target, wherein the lipid nanoparticles comprise the lipid compounds of this disclosure and the active agent (e.g., polynucleotide).

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 504,967, filed on 30 May 2023, the entirety of which application is incorporated herein by reference.

[0002] This disclosure provides lipid compounds and compositions comprising the lipid compounds of this disclosure (e.g., lipid nanoparticle (LNP) compositions). This disclosure also provides a method for delivering an active agent (e.g., polynucleotide) to target cells or tissues, e.g., extrahepatic cells or tissues, comprising administering an effective amount of the lipid nanoparticles of this disclosure to the target, wherein the lipid nanoparticles comprise the lipid compounds of this disclosure and the active agent (e.g., polynucleotide). [Background technology]

[0003] Many biologically active drugs can be difficult to deliver into cells. These active drugs include polynucleotide-based therapies (DNA-based therapies or RNA-based therapies such as mRNA or siRNA) and CRISPR / Cas9-based gene editing therapies. In recent years, lipid nanoparticles (LNPs) have been developed as encapsulation vehicles to deliver these types of biologically active drugs to cells and tissues of interest. For example, LNPs containing ionic lipids can function as vehicles to deliver biologically active drugs directly into target cells across the cell membrane and to direct the active drugs to preferred tissues of interest. [Overview of the project] [Problems that the invention aims to solve]

[0004] However, LNPs can benefit from improvements such as extended stability and cell or tissue specificity. For example, many LNPs strongly prioritize hepatocytes and tissues and have limited effectiveness in targeting extrahepatic (i.e., non-liver) cells and tissues. Accordingly, there is a need for stable LNP compositions that effectively target extrahepatic cells and tissues of a subject.

Means for Solving the Problems

[0005] The present disclosure provides, inter alia, lipid compounds and compositions (e.g., lipid nanoparticle (LNP) compositions) containing the lipid compounds of the present disclosure that can provide improved stability and / or extrahepatic targeting.

[0006] The present disclosure provides the following formula:

Chemical formula

[0007] The present disclosure provides a lipid compound of the following formula: [Chemical Formula] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 1’ each independently is (C1-C9 alkyl)-R 5 , (C2-C9 alkenyl)-R 5 , (C2-C9 alkynyl)-R 5 or (C1-C8 alkoxy)-R 5 , R 1’’ independently is (C1-C9 alkyl)-R 5 , (C2-C9 alkenyl)-R 5 , (C2-C9 alkynyl)-R 5 , (C1-C8 alkoxy)-R 5 or R 12 -R 13 , each R 5 independently is hydrogen, C1-C 12 alkyl, C2-C 12 alkenyl, C2-C 12 alkynyl, C2-C 12 alkoxy, optionally substituted C3-C 12 cycloalkyl, optionally substituted C5-C6 aryl, 1-adamantyl, 2-adamantyl, sterolyl, C(O)O-R 6 OC(O)-R 6 OC(O)O-R 6 CH(R 7 )R 8 C(O)O-CH(R 7 )R 8, C(O)O-C1~C4 alkyl-(R 9 )R 10 OC(O)-C1~C4 alkyl-(R 9 )R 10 or OC(O)CH(R 9 )R 10 And, Each R 6 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl, C7~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyl, optionally substituted C5-C6 aryl, 1-adamantyl, 2-adamantyl, or sterolyl, Each R 7 and R 8 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl or C7 ~C 12 It is an alkoxy, Each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is alkenyl, X 1 is O, NH or CHR 14 And, X 2 is O, NH or CHR 11 And, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl or C2-C 12 Alkinyl, C1~C 12 Alkoxy, (C1-C4 alkyl)-(C1-C4 alkoxy), optionally substituted C3-C 12 Cycloalkyl, (C1-C4 alkyl)-(optionally substituted C3-C 12(Cycloalkyl), optionally substituted C3-C6 heterocycle, (C1-C4 alkyl)-(optionally substituted C3-C6 heterocycle), optionally substituted C5-C6 aryl, or (C1-C4 alkyl)-(optionally substituted C5-C6 aryl), R 3 and R 4 Each of these is independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or C1-C6 hydroxyalkyl, or R 3 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R 12 These are bonded or optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl, or C1-C6 alkynyl groups. R 13 This is hydrogen, and optionally substituted C3-C 12 A cycloalkyl or optionally substituted C5-C6 aryl, R 11 is either hydrogen or a C1-C6 alkyl group, or R 11 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R 14 is either hydrogen or a C1-C6 alkyl group, or R 14 and R 2 They bond together to form optionally substituted C5-C8 cycloalkyl groups. n is 1 to 5, and m is between 1 and 4.

[0008] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 and each R 5 These are, independently, C2~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R1’’ Each of them independently is (C1~C9 alkyl)-R 5 and each R 5 These are, independently, C2~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C6~C 10 It is cycloalkyl. In certain embodiments, R 5’ is 1-adamantyl or 2-adamantyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is CH(R 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy.

[0009] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 Independently, C(O)OR 6 and each R 6 It is independently, C7~C 12 Alkyl or C7~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ Each of them independently is (C1~C9 alkyl)-R 5 And each R 5 Independently, C(O)OR 6 and each R 6 It is independently, C7~C 12 Alkyl or C7~C 12 It is Alkenil.

[0010] In a particular embodiment, R 1 , R 1’ and R1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C6~C 10 It is cycloalkyl. In certain embodiments, R 5’ It is either 1-adamantyl or 2-adamantyl.

[0011] In a particular embodiment, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ CH(R 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy.

[0012] In a particular embodiment, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ is OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0013] In a particular embodiment, R 1 and R 1’ is (C1~C9 alkyl)-R 5’ And R 5’ C(O)OR 6 and each R 6 It is independently, C7~C 12 Alkyl or C7~C 12 It is an alkenyl and R 1’ R 12 -R 13 And R 12is a bonded or optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl, or C1-C6 alkynyl, and R 13 This is hydrogen, and optionally substituted C3-C 12 It is a cycloalkyl or optionally substituted C5-C6 aryl compound.

[0014] In a particular embodiment, R 12 This is a coupling. In a particular embodiment, R 12 is a C1-C6 alkyl group. In a particular embodiment, R 12 is a C1 alkyl group. In certain embodiments, R 12 R is an optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl, or C1-C6 alkynyl. In certain embodiments, R 12 The compound is selected from pentyl, 1-methylpentyl, 4-methylpentyl, 5,5,5-trifluoropentyl, 4,4,5,5,5-pentafluoropentyl, and penta-4-inyl.

[0015] In a particular embodiment, R 13 It is hydrogen.

[0016] In a particular embodiment, R 13 C3~C are optionally substituted. 12 It is cycloalkyl. In certain embodiments, R 13 This is selected from optionally substituted cyclopropane, optionally substituted cyclobutane, and optionally substituted cyclohexane, for example, 4-pentylcyclohexyl.

[0017] In a particular embodiment, R 13 This is a condensed C3-C that has been optionally substituted. 12 Cycloalkyl, optionally substituted crosslinked C3-C 12 Cycloalkyl or optionally substituted spiro C3-C 12 It is a cycloalkyl group.

[0018] In a particular embodiment, R13 R is an optionally substituted C5-C6 aryl. In certain embodiments, R 13 This is an optionally substituted phenyl, such as 4-pentylphenyl or 3,5-di-tert-butylphenyl.

[0019] In a particular embodiment, R 13 R is an optionally substituted bicyclo[2.2.2]pentane. In certain embodiments, R 13 This is unsubstituted bicyclo[2.2.2]pentane, 1-(trifluoromethyl)bicyclo[1.1.1]pentane, or 1-methylbicyclo[1.1.1]pentane.

[0020] In a particular embodiment, R 13 R is an optionally substituted bicyclo[2.1.0]pentane. In certain embodiments, R 13 This is unsubstituted bicyclo[2.1.0]pentane.

[0021] In a particular embodiment, R 13 R is a bicyclo[3.1.0]hexane that is optionally substituted. In certain embodiments, R 13 This is 6,6-difluorobicyclo[3.1.0]hexane.

[0022] In a particular embodiment, R 13 R is a bicyclo[2.1.1]hexane that is optionally substituted. In certain embodiments, R 13 This is unsubstituted bicyclo[2.1.1]hexane or 1-fluorobicyclo[2.1.1]hexane.

[0023] In a particular embodiment, R 13 R is optionally substituted spiro[2.3]hexane. In certain embodiments, R 13 This is optionally unsubstituted spiro[2.3]hexane or 1,1-difluorospiro[2.3]hexane.

[0024] In a particular embodiment, R 13This is 1,1'-bi(cyclohexane) that has been optionally substituted.

[0025] In a particular embodiment, R 13 This is decahydronaphthalene that has been optionally substituted.

[0026] In a particular embodiment, R 13 R is a bicyclo[2.2.1]heptane that is optionally substituted. In certain embodiments, R 13 This is unsubstituted bicyclo[2.2.1]heptane or 7,7-dimethylbicyclo[2.2.1]heptane.

[0027] In a particular embodiment, R 13 R is a bicyclo[4.1.0]heptane that is optionally substituted. In certain embodiments, R 13 This is unsubstituted bicyclo[4.1.0]heptane or 7,7-difluorobicyclo[4.1.0]heptane.

[0028] In a particular embodiment, R 13 is an optionally substituted bicyclo[3.2.0]heptane. In certain embodiments, R 13 This is unsubstituted bicyclo[3.2.0]heptane.

[0029] In a particular embodiment, R 13 R is a optionally substituted spiro[3.3]heptane. In certain embodiments, R 13 This is unsubstituted spiro[3.3]heptane or 2,2-difluorospiro[3.3]heptane.

[0030] In a particular embodiment, R 13 R is an optionally substituted bicyclo[2.2.2]octane. In certain embodiments, R 13 This is unsubstituted bicyclo[2.2.2]octane or 1-methylbicyclo[2.2.2]octane.

[0031] In a particular embodiment, R 13R is an optionally substituted bicyclo[3.2.1]octane. In certain embodiments, R 13 is unsubstituted bicyclo[3.2.1]octane or 8-oxabicyclo[3.2.1]octane.

[0032] In a particular embodiment, R 13 R is optionally substituted spiro[2.5]octane. In certain embodiments, R 13 This is unsubstituted spiro[2.5]octane or 1,1-difluorospiro[2.5]octane.

[0033] In a particular embodiment, R 13 is an optionally substituted bicyclo[3.2.2]nonane. In certain embodiments, R 13 This is an unsubstituted bicyclo[3.2.2]nonane or a 1-fluorobicyclo[3.2.2]nonane.

[0034] In a particular embodiment, R 13 is an optionally substituted 1-bicyclo[3.3.1]nonane. In certain embodiments, R 13 This is unsubstituted bicyclo[3.3.1]nonane or 1-methylbicyclo[3.3.1]nonane.

[0035] In a particular embodiment, R 13 It is adamantane.

[0036] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 Independently, OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R1’’ Each of them independently is (C1~C9 alkyl)-R 5 And R 5 is OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0037] In a particular embodiment, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C(O)O-C1~C4 alkyl-(R 9 )R 10 or OC(O)-C1~C4 alkyl-(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 These are independently C(O)O-C1~C4 alkyl-(R 9 )R 10 or OC(O)-C1~C4 alkyl-(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ Each of them independently is (C1~C9 alkyl)-R 5 And R 5 C(O)O-C1~C4 alkyl-(R 9 )R 10or OC(O)-C1~C4 alkyl-(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0038] In a particular embodiment, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C6~C 10 It is a cycloalkyl, and optionally R 5’ is 1-adamantyl or 2-adamantyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C(O)OR 6 And R 6 C7~C 12 Alkyl or C7~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ CH(R 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy.

[0039] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 CH(R) 7 )R 8 And each R 7 and R 8It is independently, C7~C 12 It is an alkoxy. In certain embodiments, R 1 , R 1’ and R 1’’ Each of them independently is (C1~C9 alkyl)-R 5 And each R 5 CH(R) 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy.

[0040] In a particular embodiment, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ is OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0041] In a particular embodiment, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl or C2-C 12 It is an alkynyl. In certain embodiments, R 2 is a C4-C8 alkyl group. In a particular embodiment, R 2 This includes methyl, ethyl, propyl, isopropyl, butyl, 1-isobutyl, 2-isobutyl, tert-butyl, C5 alkyl, C6 alkyl, C8 alkyl or C 10 It is alkyl.

[0042] In a particular embodiment, R 2 C1~C 12 It is an alkoxy or (C1-C4 alkyl)-(C1-C4 alkoxy). In certain embodiments, R 2 These are methoxy, ethoxy, methoxymethyl, or ethoxyethyl.

[0043] In a particular embodiment, R 2 C3~C are optionally substituted. 12 The cycloalkyl group is a cycloalkyl group, an optionally substituted C3-C6 heterocycle, or an optionally substituted C5-C6 aryl group. In certain embodiments, R 2 This is an optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, such as 4-pentylcyclohexyl.

[0044] In a particular embodiment, R 2 R is an optionally substituted phenyl. In certain embodiments, R 2 It is 4-pentylphenyl.

[0045] In a particular embodiment, R 2 This is an optionally substituted (C1~C4 alkyl)-(optionally substituted C3~C 12 The C1-C4 alkyl group is a cycloalkyl group, a C1-C4 alkyl group with an optionally substituted C3-C6 heterocycle, or a C1-C4 alkyl group with an optionally substituted C5-C6 aryl group. In certain embodiments, R 2 These are -CH2-cyclopropyl, -(CH2)2-cyclopropyl, -CH2-cyclohexyl, -(CH2)2-cyclohexyl, -(CH2)2-(4-pentylcyclohexyl), -CH2-phenyl, or -(CH2)2-phenyl.

[0046] In a particular embodiment, R 2’ is hydrogen. In a particular embodiment, R 2’ C1~C 12 Alkyl, C2~C 12 Alkenyl or C2-C 12 It is an alkynyl. In certain embodiments, R 2 These are methoxy, ethoxy, methoxymethyl, or ethoxyethyl.

[0047] In a particular embodiment, R 2 and R 2’These combine to form optionally substituted C4-C6 cycloalkyl or C4-C6 heterocycles. In certain embodiments, R 2 and R 2’ These combine to form cyclohexane or pyran.

[0048] In a particular embodiment, X 1 This is CH2. In a particular embodiment, X 1 CHR 14 And R 14 and R 2 These combine to form optionally substituted C5-C8 cycloalkyl groups. In certain embodiments, R 14 and R 2 These combine to form optionally substituted C5 cycloalkyl or optionally substituted C6 cycloalkyl.

[0049] In a particular embodiment, X 2 is NH. In a particular embodiment, X 2 In a particular embodiment, R 3 and R 4 Each of these is independently a C1-C6 alkyl group.

[0050] In a particular embodiment, R 3 and R 4 They are bonded together to form a heterocyclic ring containing a nitrogen heteroatom. In certain embodiments, R 3 and R 4 These are bonded together to form pyrrolidine. In certain embodiments, R 3 and R 4 This combines with the aforementioned alkyl group to form quinuclidine.

[0051] In certain embodiments, n is 1, 2, 3, or 4.

[0052] In certain embodiments, m is 1, 2, 3, or 4.

[0053] In certain embodiments, p is 0, 1, 2, 3, or 4.

[0054] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C2 alkyl)-R 5 That is the case.

[0055] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C3 alkyl)-R 5 That is the case.

[0056] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C4 alkyl)-R 5 That is the case.

[0057] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C5 alkyl)-R 5 That is the case.

[0058] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C6 alkyl)-R 5 That is the case.

[0059] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C7 alkyl)-R 5 That is the case.

[0060] In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C8 alkyl)-R 5 That is the case.

[0061] In a particular embodiment, the compound is of the following formula: [ka] It is either the substance or a pharmaceutically acceptable salt thereof.

[0062] In a particular embodiment, the compound is of the following formula: [ka] It is either the substance or a pharmaceutically acceptable salt thereof.

[0063] In a particular embodiment, the compound is of the following formula: [ka] It is either the substance or a pharmaceutically acceptable salt thereof.

[0064] In certain embodiments, the compound is one of compounds 1 to 209 or a pharmaceutically acceptable salt thereof.

[0065] In certain embodiments, the compound is one of compounds 7, 8, 10, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, or 109, or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 7 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 8 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 10 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 13 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 14 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 26 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 33 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 38 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 39 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 40 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 48 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 60 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 61 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 89 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 103 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is compound 109 or a pharmaceutically acceptable salt thereof.

[0066] In certain embodiments, the Disclosure provides lipid nanoparticles comprising the compounds of the Disclosure (e.g., lipid compounds). In certain embodiments, the Disclosure provides lipid nanoparticles comprising the compounds of the Disclosure (e.g., lipid compounds), phospholipids, cholesterol, and polyethylene glycol lipids. In certain embodiments, the lipid nanoparticles comprise about 20–80 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 7.5–40 mol% of phospholipids, about 6–45 mol% of cholesterol, and about 1–4 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles comprise about 45–65 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 10 mol% of phospholipids, about 25–45 mol% of cholesterol, and about 1–4 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles comprise about 45–50 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 10 mol% of phospholipids, about 38–42 mol% of cholesterol, and about 2–3 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 47.5 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 40 mol% of cholesterol, and about 2.5 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 47.5–52.5 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 10 mol% of phospholipids, about 37–40 mol% of cholesterol, and about 1–2 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 50 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 38.5 mol% of cholesterol, and about 1.5 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 57.5–62.5 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 10 mol% of phospholipids, about 26–29 mol% of cholesterol, and about 2–3 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 60 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 27.5 mol% of cholesterol, and about 2.5 mol% of PEG lipids. In certain embodiments, the lipid nanoparticles contain about 45–50 mol% of the compounds of the Disclosure (e.g., lipid compounds), about 10 mol% of phospholipids, about 37.5–40.5 mol% of cholesterol, and about 3–4 mol% of PEG lipids.In certain embodiments, the lipid nanoparticles contain about 47.5 mol% of the compounds of the present disclosure (e.g., lipid compounds), about 39 mol% of cholesterol, and about 3.5 mol% of PEG lipids.

[0067] In certain embodiments, lipid nanoparticles contain a targeting component. In certain embodiments, the targeting component is a targeted lipid. In certain embodiments, the targeting component is an active targeting component. In certain embodiments, the active targeting component is a protein, peptide, small molecule, or antibody or its antigen-binding fragment. In certain embodiments, the active targeting component is a protein. In certain embodiments, the active targeting component is a peptide. In certain embodiments, the active targeting component is a small molecule. In certain embodiments, the active targeting component is an antibody or its antigen-binding fragment.

[0068] In certain embodiments, the lipid nanoparticles contain one or more polynucleotides encapsulated within the lipid nanoparticles. In certain embodiments, the one or more polynucleotides contain RNA. In certain embodiments, the one or more polynucleotides contain DNA. In certain embodiments, the one or more polynucleotides contain both DNA and RNA.

[0069] In certain embodiments, the Disclosure provides a pharmaceutical composition comprising lipid nanoparticles of the Disclosure and a pharmaceutically acceptable excipient. In certain embodiments, the Disclosure provides a pharmaceutical composition comprising lipid nanoparticles of the Disclosure, wherein the lipid nanoparticles comprise one or more polynucleotides encapsulated within the lipid nanoparticles and a pharmaceutically acceptable excipient.

[0070] In certain embodiments, the Disclosure provides a method for delivering polynucleotides to target cells or tissues, the method comprising administering an effective amount of the lipid nanoparticles or pharmaceutical composition of the Disclosure to target. In certain embodiments, the Disclosure provides a method for delivering polynucleotides to target cells or tissues, the method comprising administering an effective amount of the lipid nanoparticles or pharmaceutical composition of the Disclosure to target. In certain embodiments, cells or tissues include extrahepatic cells or tissues. In certain embodiments, cells or tissues include brain cells or tissues. In certain embodiments, cells or tissues include lung cells or tissues. In certain embodiments, cells or tissues include bone marrow cells or tissues. In certain embodiments, cells or tissues include spleen cells or tissues. In certain embodiments, cells or tissues include muscle cells or tissues. In certain embodiments, cells or tissues include kidney cells or tissues. In certain embodiments, cells or tissues include cardiac cells or tissues. In certain embodiments, cells or tissues include pancreatic cells or tissues. In certain embodiments, cells or tissues include immune cells or tissues.

[0071] In certain embodiments, the Disclosure provides a method for treating a disease of interest, the method comprising administering a therapeutically effective amount of the Pharmaceutical Composition of the Disclosure to the subject.

[0072] In certain embodiments, the Disclosure provides a method for producing a therapeutic composition, the method comprising encapsulating an active agent within lipid nanoparticles, the lipid nanoparticles comprising a compound of the Disclosure (e.g., a lipid compound). In certain embodiments, the active agent comprises DNA. In certain embodiments, the active agent comprises RNA. In certain embodiments, the active agent comprises both DNA and RNA.

[0073] In certain embodiments, the Disclosure provides a method for producing a vaccine or prophylactic composition, the method comprising encapsulating an active agent (e.g., RNA or DNA) within lipid nanoparticles, the lipid nanoparticles comprising a compound of the Disclosure (e.g., a lipid compound). In certain embodiments, the active agent comprises DNA. In certain embodiments, the active agent comprises RNA. In certain embodiments, the active agent comprises both DNA and RNA.

[0074] In certain embodiments, the Disclosure provides the use of the lipid nanoparticles or pharmaceutical compositions of the Disclosure in the manufacture of a pharmaceutical for delivering polynucleotides to target extrahepatic cells or tissues. In certain embodiments, the Disclosure provides the use of the lipid nanoparticles or pharmaceutical compositions of the Disclosure in the manufacture of a pharmaceutical for treating a target disease. [Brief explanation of the drawing]

[0075] [Figure 1A] This shows the 4-week stability of selective lipid nanoparticles (LNPs) prepared according to formulation F1, as measured by particle size. N / P = molar ratio of ionic nitrogen to phosphate groups. T = hours. D = days. W = weeks. [Figure 1B] This shows the 4-week stability of selected LNPs prepared according to formulation F1, as measured by the polydispersity index (PDI). N / P = molar ratio of ionic nitrogen to phosphate groups. T = hours. D = days. W = weeks. [Figure 1C] This shows the 2-week stability of selected LNPs prepared according to formulation F3, as measured by particle size. N / P = molar ratio of ionic nitrogen to phosphate groups. T = hours. D = days. W = weeks. [Figure 1D] This shows the 2-week stability of selected LNPs prepared according to formulation F3, as measured by the polydispersity index (PDI). N / P = molar ratio of ionic nitrogen to phosphate groups. T = hours. D = days. W = weeks. [Figure 2A-2D] We provide mouse organ luciferase imaging results 6 hours after IV administration of a pool of selected LNP compounds. Results are shown as total flux [p / s]. [Figure 3]We provide mouse brain luciferase imaging results 6 hours after ICV administration of a selected LNP compound pool. Results are shown as total flux [p / s]. [Figure 4A-4B] This document provides mouse lung luciferase imaging results after intratracheal administration of an LNP compound pool (compounds 2, 3, 5, and 7). Results are shown as total flux [p / s]. Figure 4A shows the in vivo imaging results, and Figure 4B shows the ex vivo imaging results. [Figure 4C] This paper provides mouse lung luciferase imaging results after intratracheal administration of an LNP compound pool (compounds 8, 9, 11, and 20). Results are presented as total flux [p / s]. [Figure 4D] This paper provides mouse lung luciferase imaging results after intratracheal administration of an LNP compound pool (compounds 12, 14, 16, and 18). Results are presented as total flux [p / s]. [Figure 5A] This document provides mouse organ luciferase imaging results after IV administration of a selected LNP compound prepared according to formulation F2. Results are expressed as total flux [p / s]. [Figure 5B] These results, normalized to MC3 activity, are shown below. [Figure 6A-6B] This document provides mouse lung and tracheal luciferase imaging results after intratracheal administration of a selected LNP compound prepared according to formulation F2. Results are shown as total flux [p / s]. Figure 6A provides in vivo imaging results, and Figure 6B provides ex vivo imaging results. [Figure 7A-7C] This document provides mouse in vivo activity screening results (mean total flux) in the spleen, femur, and muscle after administration of specific LNP compounds of this disclosure to a group of mice. [Figures 8A-8C] This document provides NHP in vivo activity screening results (proprietary molecular identification number count) in the spleen, femur, and muscle after administration of specific LNP compounds of the present disclosure to a group of cynomolgus monkeys. [Figure 9]This provides screening results (proprietary molecular identification number count) of NHP in vivo activity in the spleen after administration of specific LNP compounds of the present disclosure to a group of cynomolgus monkeys. [Figure 10] This provides NHP in vivo activity screening results (unique molecular identification number count) in the femur after administration of specific LNP compounds of the present disclosure to a group of cynomolgus monkeys. [Figure 11] This provides screening results (proprietary molecular identification number count) of NHP in vivo activity in muscle after administration of specific LNP compounds of the present disclosure to a group of cynomolgus monkeys. [Modes for carrying out the invention]

[0076] I. Lipid compounds While not bound by theory, the lipid compounds disclosed herein, when an active agent is encapsulated in an LNP containing the lipid compound, facilitate the delivery of the active agent to a desired target, such as extrahepatic cells or tissues of the subject.

[0077] In certain embodiments, this disclosure relates to formula (I): [ka] (I) The present invention provides a lipid compound or a pharmaceutically acceptable salt thereof, wherein, R 1 and R 1’ Each of them independently is (C1~C9 alkyl)-R 5 (C2~C9 Alkenil)-R 5 (C2~C9 alkynyl)-R 5 or (C1~C8 alkoxy)-R 5 And, R 1’’ These are independently (C1~C9 alkyl)-R 5 (C2~C9 Alkenil)-R 5 (C2~C9 alkynyl)-R 5 (C1~C8 alkoxy)-R 5 or R 12 -R 13 And, Each R 5 These are, independently, hydrogen, C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 Alkinyl, C2~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyls (including condensed, crosslinked, or spirocycloalkyls), optionally substituted C5-C6 aryls, 1-adamantyl, 2-adamantyl, sterolyl, C(O)OR 6 OC(O)-R 6 , OC(O)OR 6 CH(R 7 )R 8 , C(O)O-CH-(R 7 )R 8 , C(O)O-C1~C4 alkyl-(R 9 )R 10 OC(O)-C1~C4 alkyl-(R 9 )R 10 or OC(O)CH(R 9 )R 10 And, Each R 6 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl, C7~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyl (including condensed, crosslinked, or spirocycloalkyl), optionally substituted C5-C6 aryl, 1-adamantyl, 2-adamantyl, or sterolyl, Each R 7 and R 8 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl or C7 ~C 12 It is an alkoxy, Each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is alkenyl, X1 is O, NH or CHR 14 And, X 2 is O, NH or CHR 11 And, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 Alkinyl, C1~C 12 Alkoxy or (C1-C4 alkyl)-(C1-C4 alkoxy), optionally substituted C3-C 12 Cycloalkyl, (C1-C4 alkyl)-(optionally substituted C3-C 12 (Cycloalkyl), optionally substituted C3-C6 heterocycle, (C1-C4 alkyl)-(optionally substituted C3-C6 heterocycle), optionally substituted C5-C6 aryl, or (C1-C4 alkyl)-(optionally substituted C5-C6 aryl), R 2’ is hydrogen, C1~C 12 Alkyl, alkenyl, or alkynyl, C1-C 12 Alkoxy, (C1~C4 alkyl)-(C1~C4 alkoxy), R 2 and R 2’ These can combine to form optionally substituted C4-C6 cycloalkyl groups, optionally substituted C3-C6 cycloalkyl groups, or optionally substituted C3-C6 heterocycles. R 3 and R 4 Each of these is independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or C1-C6 hydroxyalkyl, or R 3 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R 11 is either hydrogen or a C1-C6 alkyl group, or R 11 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R12 These are bonded or optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl, or C1-C6 alkynyl groups. R 13 This is hydrogen, and optionally substituted C3-C 12 A cycloalkyl (including condensed, crosslinked, or spirocycloalkyl) or optionally substituted C5-C6 aryl, R 14 is either hydrogen or a C1-C6 alkyl group, or R 14 and R 2 They bond together to form optionally substituted C5-C8 cycloalkyl groups. m is 1 to 4, p is 0 to 4, and n is between 1 and 5.

[0078] In certain embodiments, this disclosure relates to formula (II): [ka] (II) The present invention provides a lipid compound or a pharmaceutically acceptable salt thereof, wherein, R 1 and R 1’ Each of them independently is (C1~C9 alkyl)-R 5 (C2~C9 Alkenil)-R 5 (C2~C9 alkynyl)-R 5 or (C1~C8 alkoxy)-R 5 And, R 1’’ These are independently (C1~C9 alkyl)-R 5 (C2~C9 Alkenil)-R 5 (C2~C9 alkynyl)-R 5 (C1~C8 alkoxy)-R 5 or R 12 -R 13 And, Each R 5 These are, independently, hydrogen, C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C12 Alkinyl, C2~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyls (including condensed, crosslinked, or spirocycloalkyls), optionally substituted C5-C6 aryls, 1-adamantyl, 2-adamantyl, sterolyl, C(O)OR 6 OC(O)-R 6 , OC(O)OR 6 CH(R 7 )R 8 , C(O)O-CH(R 7 )R 8 , C(O)O-C1~C4 alkyl-(R 9 )R 10 OC(O)-C1~C4 alkyl-(R 9 )R 10 or OC(O)CH(R 9 )R 10 And, Each R 6 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl, C7~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyl (including condensed, crosslinked, or spirocycloalkyl), optionally substituted C5-C6 aryl, 1-adamantyl, 2-adamantyl, or sterolyl, Each R 7 and R 8 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl or C7 ~C 12 It is an alkoxy, Each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is alkenyl, X 1 is O, NH or CHR 14 And, X 2 is O, NH or CHR 11And, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 Alkinyl, C1~C 12 Alkoxy or (C1-C4 alkyl)-(C1-C4 alkoxy), optionally substituted C3-C 12 Cycloalkyl, (C1-C4 alkyl)-(optionally substituted C3-C 12 (Cycloalkyl), optionally substituted C3-C6 heterocycle, (C1-C4 alkyl)-(optionally substituted C3-C6 heterocycle), optionally substituted C5-C6 aryl, or (C1-C4 alkyl)-(optionally substituted C5-C6 aryl), R 3 and R 4 Each of these is independently hydrogen, C1-C6 alkyl, C1-C6 alkoxy or C1-C6 hydroxyalkyl, or R 3 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R 11 is either hydrogen or a C1-C6 alkyl group, or R 11 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R 12 These are bonded or optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl, or C1-C6 alkynyl groups. R 13 This is hydrogen, and optionally substituted C3-C 12 A cycloalkyl (including condensed, crosslinked, or spirocycloalkyl) or optionally substituted C5-C6 aryl, R 14 is either hydrogen or a C1-C6 alkyl group, or R 14 and R 2 They bond together to form optionally substituted C5-C8 cycloalkyl groups. n is 1 to 5, and m is between 1 and 4.

[0079] In a particular embodiment, R 1 , R 1’ and R 1’’ Each of them independently is (C1~C9 alkyl)-R 5 (C2~C9 Alkenil)-R 5 (C2~C9 alkynyl)-R 5 or (C1~C8 alkoxy)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three are the same. In a particular embodiment, R 1 , R 1’ and R 1’’ These two are the same. In a particular embodiment, R 1 , R 1’ and R 1’’ All three are different.

[0080] In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C2 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C2 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C2 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C3 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C3 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’All three of these are (C3 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C4 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C4 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C4 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C5 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C5 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C5 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C6 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C6 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C6 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C7 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R1’’ These two are independently (C7 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C8 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C8 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ These two are independently (C8 alkyl)-R 5 In a particular embodiment, R 1 , R 1’ and R 1’’ All three of these are (C8 alkyl)-R 5 That is the case.

[0081] In a specific method of operation, each R 5 These are, independently, hydrogen, C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 Alkinyl, C2~C 12 Alkoxy, optionally substituted C3-C 12 Cycloalkyls (including condensed, crosslinked, or spirocycloalkyls), optionally substituted C5-C6 aryls, 1-adamantyl, 2-adamantyl, sterolyl, C(O)OR 6 OC(O)-R 6 , OC(O)OR 6 CH(R 7 )R 8 , C(O)O-CH(R 7 )R 8 , C(O)O-C1~C4 alkyl-(R 9 )R 10 OC(O)-C1~C4 alkyl-(R 9 )R 10 or OC(O)CH(R 9 )R 10 In a particular embodiment, R 5All of the bases are the same. In a particular embodiment, two R 5 The base is the same. In a particular embodiment, R 5 Everything is fundamentally different.

[0082] In a specific method of operation, each R 6 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl, C7~C 12 Alkoxy, optionally substituted C3-C 12 The compounds are cycloalkyl (including condensed, crosslinked, or spirocycloalkyl), optionally substituted C5-C6 aryls, 1-adamantyl, 2-adamantyl, or sterolyl. In certain embodiments, each R 7 and R 8 It is independently, C7~C 12 Alkyl, C7~C 12 Alkenil, C7~C 12 Alkinyl or C7 ~C 12 It is an alkoxy. In a particular embodiment, each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0083] In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C1~C9 alkyl)-R 5 And R 5 C2~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 These are, independently, C2~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ All three of these are independently (C1~C9 alkyl)-R5 And each R 5 These are, independently, C2~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C6~C 10 It is cycloalkyl. In certain embodiments, R 5’ is 1-adamantyl or 2-adamantyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is CH(R 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy.

[0084] In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C1~C9 alkyl)-R 5 And R 5 C(O)OR 6 And R 6 C7~C 12 Alkyl or C7~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 Independently, C(O)OR 6 And each R 6 It is independently, C7~C 12 Alkyl or C7~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ All three of these are independently (C1~C9 alkyl)-R 5 And each R 5 Independently, C(O)OR 6And each R 6 It is independently, C7~C 12 Alkyl or C7~C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ C6~C 10 It is cycloalkyl. In certain embodiments, R 5’ is 1-adamantyl or 2-adamantyl. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ CH(R 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ is OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0085] In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C1~C9 alkyl)-R 5 And R 5 is OC(O)CH(R 9 )R 10 And R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is an alkenyl. In certain embodiments, R 1, R 1’ and R 1’’ Two of them are independently (C1-C9 alkyl)-R 5 Each R 5 is independently OC(O)CH(R 9 )R 10 Each R 9 and R 10 are independently C1-C 12 alkyl or C2-C 12 alkenyl. In certain embodiments, all three of R 1 , R 1’ and R 1’’ are independently (C1-C9 alkyl)-R 5 Each R 5 is OC(O)CH(R 9 )R 10 Each R 9 and R 10 are independently C1-C 12 alkyl or C2-C 12 alkenyl. In certain embodiments, one of R 1 , R 1’ and R 1’’ is (C1-C9 alkyl)-R 5’ R 5’ is C6-C 10 cycloalkyl (e.g., R 5’ is 1-adamantyl or 2-adamantyl). In certain embodiments, one of R 1 , R 1’ and R[[ID=6l]] 1’’ is (C1-C9 alkyl)-R 5’ R 5’ is C(O)O-R 6 R 6 is C7-C 12 alkyl or C7-C 12 alkenyl. In certain embodiments, one of R 1 , R 1’ and R<000B900>is (C1-C9 alkyl)-R 5’ R<000090Z>is CH(R 7 )R 8 Each R 7 [ and R8 is independently C7-C 12 alkoxy.

[0086] In certain embodiments, R 1 and R 1’ are (C1-C9 alkyl)-R 5’ where R 5’ is C(O)O-R 6 and each R 6 is independently C7-C 12 alkyl or C7-C 12 alkenyl, and R 1’ is R 12 -R 13 where R 12 is a bond or optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl or C1-C6 alkynyl, and R 13 is hydrogen, optionally substituted C3-C 12 cycloalkyl (including fused, bridged or spirocycloalkyl) or optionally substituted C5-C6 aryl.

[0087] [[ID=�9]]In certain embodiments, R 12 is a bond.

[0088] In certain embodiments, R 12 is optionally substituted branched or unbranched C1-C6 alkyl, C1-C6 alkenyl or C1-C6 alkynyl. In certain embodiments, R 12 is C1 alkyl. In certain embodiments, R 12 is optionally substituted branched C1-C6 alkyl. In certain embodiments, R 12 is pentyl. In certain embodiments, R 12 is 1-methylpentyl. In certain embodiments, R 12 is 4-methylpentyl. In certain embodiments, R 12 is 5,5,5-trifluoropentyl. In certain embodiments, R 12 is 4,4,5,5,5-pentafluoropentyl. In certain embodiments, R12 R is a branched C1-C6 alkynyl that is optionally substituted. In certain embodiments, R 12 It is penta-4-inyl.

[0089] In a particular embodiment, R 13 It is hydrogen.

[0090] In a particular embodiment, R 13 C3~C are optionally substituted. 12 It is a cycloalkyl (including condensed, crosslinked, or spirocycloalkyl). In certain embodiments, R 13 is an optionally substituted cyclopropane. In certain embodiments, R 13 is an optionally substituted cyclobutane. In certain embodiments, R 13 R is an optionally substituted cyclohexane. In certain embodiments, R 13 is 4-pentylcyclohexyl. In certain embodiments, R 13 R is an optionally substituted C5-C6 aryl. In certain embodiments, R 13 R is an optionally substituted phenyl. In certain embodiments, R 13 is 4-pentylphenyl. In certain embodiments, R 13 It is 3,5-di-tert-butylphenyl.

[0091] In a particular embodiment, R 13 This is a condensed C3-C that has been optionally substituted. 12 It is cycloalkyl. In certain embodiments, R 13 This is a selectively substituted cross-linked C3~C 12 It is cycloalkyl. In certain embodiments, R 13 This is a spiro C3-C that has been selectively substituted. 12 It is cycloalkyl. In certain embodiments, R 13R is an optionally substituted bicyclo[2.2.2]pentane, such as unsubstituted bicyclo[2.2.2]pentane, 1-(trifluoromethyl)bicyclo[1.1.1]pentane, or 1-methylbicyclo[1.1.1]pentane. In certain embodiments, R 13 R is an optionally substituted bicyclo[2.1.0]pentane, for example, an unsubstituted bicyclo[2.2.2]pentane. In certain embodiments, R 13 R is an optionally substituted bicyclo[3.1.0]hexane, for example, unsubstituted bicyclo[3.1.0]hexane or 6,6-difluorobicyclo[3.1.0]hexane. In certain embodiments, R 13 R is an optionally substituted bicyclo[2.1.1]hexane, for example, unsubstituted bicyclo[2.1.1]hexane or 1-fluorobicyclo[2.1.1]hexane. In certain embodiments, R 13 R is an optionally substituted spiro[2.3]hexane, for example, unsubstituted spiro[2.3]hexane or 1,1-difluorospiro[2.3]hexane. In certain embodiments, R 13 is optionally substituted 1,1'-bi(cyclohexane). In certain embodiments, R 13 is an optionally substituted decahydronaphthalene. In certain embodiments, R 13 R is an optionally substituted bicyclo[2.2.1]heptane, for example, unsubstituted bicyclo[2.2.1]heptane or 7,7-dimethylbicyclo[2.2.1]heptane. In certain embodiments, R 13 R is an optionally substituted bicyclo[4.1.0]heptane, for example, unsubstituted bicyclo[4.1.0]heptane or 7,7-difluorobicyclo[4.1.0]heptane. In certain embodiments, R 13 R is an optionally substituted bicyclo[3.2.0]heptane, for example, an unsubstituted bicyclo[3.2.0]heptane. In certain embodiments, R 13 R is an optionally substituted spiro[3.3]heptane, for example, unsubstituted spiro[3.3]heptane or 2,2-difluorospiro[3.3]heptane. In certain embodiments, R 13is optionally substituted bicyclo[2.2.2]octane, such as unsubstituted bicyclo[2.2.2]octane or 1-methylbicyclo[2.2.2]octane. In certain embodiments, R 13 is optionally substituted bicyclo[3.2.1]octane, such as unsubstituted bicyclo[3.2.1]octane or 8-oxabicyclo[3.2.1]octane. In certain embodiments, R 13 is optionally substituted spiro[2.5]octane, such as unsubstituted spiro[2.5]octane or 1,1-difluorospiro[2.5]octane. In certain embodiments, R 13 is optionally substituted bicyclo[3.2.2]nonane, such as unsubstituted bicyclo[3.2.2]nonane or 1-fluorobicyclo[3.2.2]nonane. In certain embodiments, R 13 is optionally substituted bicyclo[3.3.1]nonane, such as unsubstituted bicyclo[3.3.1]nonane or 1-methylbicyclo[3.3.1]nonane. In certain embodiments, R 13 is adamantane.

[0092] In certain embodiments, R 1 , R 1’ and R 1’’ at least one of is (C1-C9 alkyl)-R 5 , where R 5 is CH(R 7 )R 8 , and R 7 and R 8 are independently C7-C 12 alkoxy. In certain embodiments, two of R 1 , R 1’ and R 1’’ are independently (C_{1}-C_{9}alkyl)-R 5 , each R 5 is independently CH(R 7 )R 8 , and each R 7 and R 8 are independently C7-C 12 4 alkoxy. In certain embodiments, R 1 , R1’ and R 1’’ Each of them independently is (C1~C9 alkyl)-R 5 And each R 5 CH(R) 7 )R 8 And each R 7 and R 8 It is independently, C7~C 12 It is an alkoxy. In certain embodiments, R 1 , R 1’ and R 1’’ One of them is (C1~C9 alkyl)-R 5’ And R 5’ is OC(O)CH(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0093] In a particular embodiment, R 1 , R 1’ and R 1’’ At least one of them is (C1~C9 alkyl)-R 5 And R 5 C(O)O-C1~C4 alkyl-(R 9 )R 10 or OC(O)-C1~C4 alkyl-(R 9 )R 10 And each R 9 and R 10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is an alkenyl. In certain embodiments, R 1 , R 1’ and R 1’’ These two are independently (C1~C9 alkyl)-R 5 And each R 5 These are independently C(O)O-C1~C4 alkyl-(R 9 )R 10 or OC(O)-C1~C4 alkyl-(R 9 )R 10 And each R 9 and R10 These are, independently, C1~C 12 Alkyl or C2-C 12 It is Alkenil.

[0094] In a particular embodiment, X 1 O, NH, CHR 14 And R 14 is hydrogen or a C1-C6 alkyl group. In certain embodiments, X 1 This is CH2. In a particular embodiment, X 1 is NH. In a particular embodiment, X 1 In a particular embodiment, X 1 CHR 14 And R 14 and R 2 These combine to form optionally substituted C5-C8 cycloalkyl groups. In certain embodiments, R 14 and R 2 They bond together to form an optionally substituted C5 cycloalkyl. In certain embodiments, R 14 and R 2 These atoms bond together to form optionally substituted C6 cycloalkyl groups.

[0095] In a particular embodiment, X 2 is O, NH or CHR 11 And R 11 is hydrogen or a C1-C6 alkyl group. In certain embodiments, X 2 CHR 11 And R 11 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. In certain embodiments, R 11 and R 4 They bind together to form 1-methylpyrrolidine (R 3 It forms a methyl group. In certain embodiments, R 11 and R 4 It binds together with 1-methylpiperidine (R 3 It forms a structure that has a methyl group.

[0096] In a particular embodiment, X 2 is NH. In a particular embodiment, X 2 It is O.

[0097] In a particular embodiment, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 Alkinyl, C1~C 12 Alkoxy, (C1-C4 alkyl)-(C1-C4 alkoxy), optionally substituted C3-C 12 Cycloalkyl, (C1-C4 alkyl)-(optionally substituted C3-C 12 These are cycloalkyl groups, optionally substituted C3-C6 heterocycles, (C1-C4 alkyl)-(optionally substituted C3-C6 heterocycles), optionally substituted C5-C6 aryl groups, or (C1-C4 alkyl)-(optionally substituted C5-C6 aryl groups).

[0098] In a particular embodiment, R 2 C1~C 12 Alkyl, C2~C 12 Alkenyl, C2~C 12 It is an alkynyl. In certain embodiments, R 2 C1~C 12 It is alkyl. In certain embodiments, R 2 is a C4-C8 alkyl group. In a particular embodiment, R 2 is methyl. In certain embodiments, R 2 is trifluoromethyl. In a particular embodiment, R 2 is ethyl. In certain embodiments, R 2 is propyl or isopropyl. In certain embodiments, R 2 is butyl, 1-isobutyl, 2-isobutyl, or tert-butyl. In certain embodiments, R 2 is a C5 alkyl group. In certain embodiments, R 2 is a C6 alkyl group. In certain embodiments, R 2is a C8 alkyl group. In certain embodiments, R 2 C 10 It is alkyl.

[0099] In a particular embodiment, R 2 C1~C 12 It is an alkoxy. In certain embodiments, R 2 is (C1-C4 alkyl)-(C1-C4 alkoxy). In certain embodiments, R 2 R is methoxymethyl. In certain embodiments, R 2 It is ethoxyethyl.

[0100] In a particular embodiment, R 2 C3~C are optionally substituted. 12 The cycloalkyl group is a cycloalkyl group, an optionally substituted C3-C6 heterocycle, or an optionally substituted C5-C6 aryl group. In certain embodiments, R 2 is an optionally substituted cyclopropyl. In certain embodiments, R 2 is optionally substituted cyclobutyl. In certain embodiments, R 2 is an optionally substituted cyclopentyl. In certain embodiments, R 2 is an optionally substituted cyclohexyl. In certain embodiments, R 2 is 4-pentylcyclohexyl. In certain embodiments, R 2 R is an optionally substituted phenyl. In certain embodiments, R 2 It is 4-pentylphenyl.

[0101] In a particular embodiment, R 2 (C1~C4 alkyl)-(C3~C optionally substituted) 12 The C1-C4 alkyl group is a cycloalkyl group, a C1-C4 alkyl group with an optionally substituted C3-C6 heterocycle, or a C1-C4 alkyl group with an optionally substituted C5-C6 aryl group. In certain embodiments, R 2These are -CH2-cyclopropyl, -(CH2)2-cyclopropyl, -CH2-cyclohexyl, -(CH2)2-cyclohexyl, -CH2-phenyl, or -(CH2)2-phenyl.

[0102] In a particular embodiment, R 2’ is hydrogen. In a particular embodiment, R 2’ C1~C 12 Alkyl, C2~C 12 Alkenyl or C2-C 12 It is an alkynyl. In certain embodiments, R 2’ is methyl. In certain embodiments, R 2’ is ethyl. In certain embodiments, R 2’ is propyl. In certain embodiments, R 2’ is butyl. In a particular embodiment, R 2’ is a C5 alkyl group. In certain embodiments, R 2’ is a C6 alkyl group. In certain embodiments, R 2’ C1~C 12 It is an alkoxy. In certain embodiments, R 2’ is (C1-C4 alkyl)-(C1-C4 alkoxy). In certain embodiments, R 2’ R is methoxymethyl. In certain embodiments, R 2’ R is ethoxyethyl. In certain embodiments, R 2’ R 2 It is the same as this.

[0103] In a particular embodiment, R 2 and R 2’ These combine to form optionally substituted C4-C6 cycloalkyl groups. In certain embodiments, R 2 and R 2’ These combine to form optionally substituted cyclohexane. In certain embodiments, R 2 and R 2’ These combine to form a C4-C6 heteroring with optional substitutions. In certain embodiments, R 2 and R 2’These combine to form arbitrarily substituted pyran.

[0104] In a particular embodiment, R 3 and R 4 Each of these is independently hydrogen, a C1-C6 alkyl group, a C1-C6 alkoxy group, or a C1-C6 hydroxyalkyl group. In certain embodiments, R 3 and R 4 Each of these is independently a C1-C6 alkyl group. In a particular embodiment, R 3 and R 4 They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. In certain embodiments, R 3 and R 4 They bond together to form a heterocyclic ring containing a nitrogen heteroatom. In certain embodiments, R 3 and R 4 These combine to form pyrrolidine. In certain embodiments, R 3 and R 4 It combines with the aforementioned alkyl group to form quinuclidine.

[0105] In certain embodiments, n is 1 to 5. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5.

[0106] In certain embodiments, m is 1 to 4. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. In certain embodiments, m is 4.

[0107] In certain embodiments, p is between 0 and 4. In certain embodiments, p is 0. In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, p is 4.

[0108] Examples of lipid compounds of formulas (I) and (II) are shown in Table 1. This also includes pharmaceutically acceptable salts of the compounds shown in Table 1.

[0109] [Table 1]

[0110] [Table 2]

[0111] [Table 3]

[0112] [Table 4]

[0113] [Table 5]

[0114] [Table 6]

[0115] [Table 7]

[0116] [Table 8]

[0117] [Table 9]

[0118] [Table 10]

[0119] Table 11

[0120] Table 12

[0121] Table 13

[0122] Table 14

[0123] Table 15

[0124] Table 16

[0125] Table 17

[0126] Table 18

[0127] Table 19

[0128] Table 20

[0129] Table 21

[0130] Table 22

[0131] Table 23

[0132] Table 24

[0133] Table 25

[0134] Table 26

[0135] Table 27

[0136] Table 28

[0137] Table 29

[0138] Table 30

[0139] Table 31

[0140] Table 32

[0141] Table 33

[0142] Table 34

[0143] Table 35

[0144] Table 36

[0145] Table 37

[0146] Table 38

[0147] Table 39

[0148] Table 40

[0149] Table 41

[0150] Table 42

[0151] Table 43

[0152] Table 44

[0153] Table 45

[0154] Table 46

[0155] Table 47

[0156] Table 48

[0157] Table 49

[0158] Table 50

[0159] Table 51

[0160] Table 52

[0161] Table 53

[0162] Table 54

[0163] Table 55

[0164] Table 56

[0165] Table 57

[0166] Table 58

[0167] Table 59

[0168] Table 60

[0169] Table 61

[0170] Table 62

[0171] [Table 63]

[0172] [Table 64]

[0173] [Table 65]

[0174] [Table 66]

[0175] [Table 67]

[0176] [Table 68]

[0177] [Table 69]

[0178] [Table 70]

[0179] In certain embodiments, the lipid compound is one of compounds 1 to 209.

[0180] In certain embodiments, the lipid compound is one of compounds 7, 8, 10, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, or 109, or a pharmaceutically acceptable salt thereof.

[0181] In certain embodiments, the lipid compound is one of compounds 1 to 20 or a pharmaceutically acceptable salt thereof.

[0182] In certain embodiments, the lipid compound is compound 14 or a pharmaceutically acceptable salt thereof.

[0183] In certain embodiments, the lipid compound is one of compounds 7, 8, 10, 13, or 18, or a pharmaceutically acceptable salt thereof.

[0184] In certain embodiments, the lipid compound is one of compounds 1-4 or 6-12 or a pharmaceutically acceptable salt thereof.

[0185] In certain embodiments, the lipid compound is one of compounds 17, 60, or 61, or a pharmaceutically acceptable salt thereof.

[0186] In certain embodiments, the lipid compound is one of compounds 50 to 58 or a pharmaceutically acceptable salt thereof.

[0187] In certain embodiments, the lipid compound is one of compounds 31, 41-43, or 47, or a pharmaceutically acceptable salt thereof.

[0188] In certain embodiments, the lipid compound is one of compounds 24, 25, 28, 29, 33-36, 38, or 40, or a pharmaceutically acceptable salt thereof.

[0189] In certain embodiments, the lipid compound is one of compounds 62-34 or 66 or a pharmaceutically acceptable salt thereof.

[0190] In certain embodiments, the lipid compound is one of compounds 21, 23, 26, 27, 37, 39, or 44-46, or a pharmaceutically acceptable salt thereof.

[0191] In certain embodiments, the lipid compound is one of compounds 13-14, 67, 73, 81, 91, 93, 96, or 98-101, or a pharmaceutically acceptable salt thereof.

[0192] In certain embodiments, the lipid compound is any one of compounds 14-15, 49-50, 59, or 102-103, or a pharmaceutically acceptable salt thereof.

[0193] In certain embodiments, the lipid compound is one of compounds 8, 14, 7, 10, or 60-61, or a pharmaceutically acceptable salt thereof.

[0194] In certain embodiments, the lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof is an ionic lipid compound.

[0195] II. Lipid Compositions Lipid nanoparticles (LNPs) In certain embodiments, the disclosure provides lipid compositions (e.g., lipid nanoparticles (LNPs)) comprising at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the disclosure provides lipid nanoparticles (LNPs) comprising at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid nanoparticles are microspherical vesicles (i.e., liposomes) comprising a liposome lamellar phase lipid bilayer (monolayer or multilayer) enclosing an interlayer space. In certain embodiments, the lipid nanoparticles are nanospherical vesicles comprising a liposome lamellar phase lipid bilayer (monolayer or multilayer).

[0196] In certain embodiments, the lipid nanoparticles (LNPs) comprise (i) at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) at least one phospholipid (e.g., distearoylphosphatidylcholine (DSPC)), (iii) at least one helper lipid (e.g., cholesterol), and (iv) at least one PEG lipid. In certain embodiments, the LNPs comprise (i) at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) at least one phospholipid (e.g., distearoylphosphatidylcholine (DSPC)), (iii) at least one cholesterol, and (iv) at least one PEG lipid. In certain embodiments, the lipid nanoparticles (LNPs) comprise one or more additional lipid components.

[0197] In certain embodiments, the lipid nanoparticles (LNPs) comprise a lipid compound selected from any one of compounds 1 to 209. In certain embodiments, the lipid nanoparticles (LNPs) comprise a lipid compound selected from any one of compounds 7, 8, 10, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, or 109 or a pharmaceutically acceptable salt thereof.

[0198] The phospholipids for use in the lipid compositions of this disclosure (e.g., LNPs) may be neutral, uncharged, or zwitterionic phospholipids. Examples of phospholipids for use in lipid compositions include dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), phosphocholine (DOPC), dimyristoylphosphatidylcholine (DMPC), phosphatidylcholine (PLPC), l,2-distearoyl-sn-glycero-3-phosphocholine (DAPC), phosphatidylethanolamine (PE), egg phosphatidylcholine (EPC), dilaurylphosphatidylcholine (DLPC), dimyristoylphosphatidylcholine (DMPC), l-myristoyl-2-palmitoylphosphatidylcholine (MPPC), l-palmitoyl-2-myristoylphosphatidylcholine (PMPC), l-palmitoyl-2-stearoylphosphatidylcholine (PSPC), l,2 Examples of phospholipids include, but are not limited to, diarachidoyl-sn-glycero-3-phosphocholine (DBPC), 1-stearoyl-2-palmitoylphosphatidylcholine (SPPC), l,2-dieicosenoyl-sn-glycero-3-phosphocholine (DEPC), palmitoyloleoylphosphatidylcholine (POPC), lysophosphatidylcholine, dioleoylphosphatidylethanolamine (DOPE), dilinoleoylphosphatidylcholine distearoylphosphatidylethanolamine (DSPE), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), palmitoyloleoylphosphatidylethanolamine (POPE), lysophosphatidylethanolamine, and combinations thereof. In certain embodiments, the phospholipid is distearoylphosphatidylcholine (DSPC).

[0199] Helper lipids for use in lipid compositions (e.g., LNPs) of this disclosure include steroids, sterols, and alkylresorcinols. Examples of helper lipids for use in lipid compositions include, but are not limited to, cholesterol, cholesterol hemisuccinate, and 5-heptadecylresorcinol. In certain embodiments, the helper lipid is cholesterol.

[0200] The PEG lipids for use in lipid compositions (e.g., LNPs) of this disclosure include compounds comprising a lipid moiety bonded to a PEG-based polymer moiety (i.e., a PEG moiety). In certain embodiments, the lipid moiety of the PEG lipid is delivered from a diacylglycerol or diacylglycamide group. In certain embodiments, the lipid moiety of the PEG lipid is delivered from a dialkylglycerol or dialkylglycamide group. In certain embodiments, the dialkylglycerol or dialkylglycamide group has an alkyl chain length of saturated or unsaturated carbon atoms from about C4 to about C40. In certain embodiments, the alkyl chain length is about C 10 It is approximately C20. In certain embodiments, the dialkylglycerol or dialkylglycamide group comprises one or more functional groups (e.g., amide or ester). In certain embodiments, the dialkylglycerol or dialkylglycamide group comprises one or more substituted alkyl groups.

[0201] The PEG portion of the PEG lipid may contain any polyethylene glycol (PEG) or other polyalkylene ether polymer, including linear or branched polymers optionally substituted with ethylene glycol or ethylene oxide. The PEG portion may have a molecular weight of approximately 130 to approximately 50,000 Da, approximately 150 to approximately 25,000 Da, approximately 150 to approximately 15,000 Da, approximately 150 to approximately 10,000 Da, approximately 150 to approximately 5,000 Da, approximately 150 to approximately 4,000 Da, approximately 150 to approximately 3,000 Da, approximately 150 to approximately 2,500 Da, approximately 150 to approximately 2,000 Da, approximately 500 to approximately 3,000 Da, approximately 500 to approximately 2,000 Da, approximately 1,000 to approximately 3,000 Da, or approximately 1,000 to approximately 2,000 Da. In certain embodiments, the PEG portion includes PEG2000 (having 2,000 Da).

[0202] In certain embodiments, the lipid composition (e.g., LNP) contains about 40 to 70 mol% of at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 20 to 80 mol% of the lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 45 to 65 mol% of the lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 45 to 50 mol% (e.g., about 47.5 mol%) of the lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 47.5 to 52.5 mol% (e.g., about 50 mol%) of the lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 50 to 55 mol% (e.g., about 52.5 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 52.5 to 57.5 mol% (e.g., about 55 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 55 to 60 mol% (e.g., about 57.5 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 57.5 to 62.5 mol% (e.g., about 60 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 60 to 65 mol% (e.g., about 62.5 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the lipid composition contains about 62.5 to 67.5 mol% (e.g., about 65 mol%) of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

[0203] In certain embodiments, the lipid composition (e.g., LNP) contains about 7.5 to 40 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)). In certain embodiments, the lipid composition (e.g., LNP) contains about 7.5 to 12.5 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)). In certain embodiments, the lipid composition contains about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)).

[0204] In certain embodiments, the lipid composition (e.g., LNP) contains about 6 to 50 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition (e.g., LNP) contains about 20 to 50 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition (e.g., LNP) contains about 6 to 45 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 25 to 45 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 26 to 29 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 27.5 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 37 to 40 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 38.5 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 37.5 to 40.5 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 39 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 38 to 42 mol% of helper lipids (e.g., cholesterol). In certain embodiments, the lipid composition contains about 40 mol% of helper lipids (e.g., cholesterol).

[0205] In certain embodiments, the lipid composition (e.g., LNP) contains about 1 to 4 mol% of PEG lipids. In certain embodiments, the lipid composition contains about 1 to 2 mol% (e.g., 1.5 mol%) of PEG lipids. In certain embodiments, the lipid composition contains about 1.5 to 2.5 mol% (e.g., 2 mol%) of PEG lipids. In certain embodiments, the lipid composition contains about 2 to 3 mol% (e.g., 2.5 mol%) of PEG lipids. In certain embodiments, the lipid composition contains about 2.5 to 3.5 mol% (e.g., 3 mol%) of PEG lipids. In certain embodiments, the lipid composition contains about 3 to 4 mol% (e.g., 3.5 mol%) of PEG lipids.

[0206] In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 20 to 80 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 7.5 to 40 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 6 to 45 mol% of helper lipids (e.g., cholesterol), and (iv) about 1 to 4 mol% of PEG lipids. In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 40 to 70 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, and (ii) about 7.5 to 12 (iii) about 0.5 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 20-50 mol% of helper lipids (e.g., cholesterol), and (iv) about 1-4 mol% of PEG lipids. In certain embodiments, the lipid composition comprises (i) about 45-65 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 25-45 mol% of helper lipids (e.g., cholesterol), and (iv) about 1-4 mol% of PEG lipids.

[0207] In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 45-50 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 38-42 mol% of helper lipids (e.g., cholesterol), and (iv) about 2-3 mol% of PEG lipids. In certain embodiments, the lipid composition comprises (i) about 47.5 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 40 mol% of helper lipids (e.g., cholesterol), and (iv) about 2.5 mol% of PEG lipids.

[0208] In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 47.5 to 52.5 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 37 to 40 mol% of helper lipids (e.g., cholesterol), and (iv) about 1 to 2 mol% of PEG lipids. In certain embodiments, the lipid composition comprises (i) about 50 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 38.5 mol% of helper lipids (e.g., cholesterol), and (iv) about 1.5 mol% of PEG lipids.

[0209] In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 57.5 to 62.5 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 26 to 29 mol% of helper lipids (e.g., cholesterol), and (iv) about 2 to 3 mol% of PEG lipids. In certain embodiments, the lipid composition comprises (i) about 60 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 27.5 mol% of helper lipids (e.g., cholesterol), and (iv) about 2.5 mol% of PEG lipids.

[0210] In certain embodiments, the lipid composition (e.g., LNP) comprises (i) about 45-50 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 37.5-40.5 mol% of helper lipids (e.g., cholesterol), and (iv) about 3-4 mol% of PEG lipids. In certain embodiments, the lipid composition comprises (i) about 47.5 mol% of a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, (ii) about 10 mol% of phospholipids (e.g., distearoylphosphatidylcholine (DSPC)), (iii) about 39 mol% of helper lipids (e.g., cholesterol), and (iv) about 3.5 mol% of PEG lipids.

[0211] In certain embodiments, the lipid composition of the Disclosure (e.g., LNP) further comprises a targeting component. In certain embodiments, the targeting component is a passive targeting component. In certain embodiments, the passive targeting component comprises a targeting lipid. In certain embodiments, the targeting lipid comprises at least one cationic targeting lipid. Examples of cationic targeting lipids for use in the lipid composition of the Disclosure (e.g., LNP) include, but are not limited to, 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and N-[1(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA). In certain embodiments, the targeting lipid comprises at least one anionic targeting lipid. Examples of anionically targeted lipids for use in the lipid compositions (e.g., LNPs) of this disclosure include, but are not limited to, phosphatidic acid (PA), bis(monoacylglycero)phosphate (BMP), hemibis(monoacylglycero)phosphate (hemiBMP), and bis(diacylglycero)phosphate (BDP).

[0212] In certain embodiments, the targeting component of the lipid composition disclosed herein (e.g., lipid nanoparticles (LNPs)) is an active targeting component. In certain embodiments, the active targeting component includes a protein. In certain embodiments, the active targeting component includes a peptide. In certain embodiments, the active targeting component includes a small molecule. In certain embodiments, the active targeting component includes an antibody. In certain embodiments, the active targeting component includes an antigen-binding fragment of an antibody.

[0213] Active drug In certain embodiments, the lipid compositions disclosed herein (e.g., lipid nanoparticles (LNPs)) include at least one active agent (e.g., RNA, DNA). In certain embodiments, the disclosure provides lipid compositions (e.g., lipid nanoparticles (LNPs)) comprising at least one lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, and at least one active agent (e.g., RNA, DNA) in the lipid composition, for example, in the LNPs.

[0214] In certain embodiments, the active agent comprises one or more polynucleotides. In certain embodiments, the active agent comprises one or more RNAs. In certain embodiments, the active agent comprises one or more DNAs. In certain embodiments, the active agent comprises one or more RNAs and one or more DNAs.

[0215] In certain embodiments, the active agent comprises mRNA. In certain embodiments, the active agent comprises mRNA encoding an RNA-inducing DNA binding agent (e.g., a Cas nuclease such as Cas9).

[0216] In certain embodiments, the active agent includes gRNA. In certain embodiments, the active agent includes dgRNA or sgRNA.

[0217] In certain embodiments, the active agent includes inhibitory polynucleotides, such as siRNA (i.e., non-coding small interfering RNA molecules).

[0218] Compositions and methods In certain embodiments, the Disclosure provides a pharmaceutical composition comprising the lipid composition of the Disclosure (e.g., lipid nanoparticles (LNPs)). In certain embodiments, the pharmaceutical composition comprises the lipid composition of the Disclosure (e.g., lipid nanoparticles (LNPs)) and at least one pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition is an aqueous solution or a turbidity. In certain embodiments, the pharmaceutical composition is an oil-based solution or a turbidity.

[0219] In certain embodiments, the Disclosure provides a method for delivering an active agent (e.g., polynucleotide) to target cells or tissues (e.g., extrahepatic cells or tissues). In certain embodiments, the method includes administering an effective amount of the lipid composition of the Disclosure (e.g., LNP) to a target. In certain embodiments, the method includes administering a pharmaceutical composition comprising an effective amount of the lipid composition of the Disclosure (e.g., LNP) to a target.

[0220] In certain embodiments, the Disclosure describes compositions of the Disclosure for use in the delivery of an active agent (e.g., polynucleotide) to target cells or tissues (e.g., extrahepatic cells or tissues). In certain embodiments, the Disclosure describes compositions of the Disclosure for use in the delivery of an active agent (e.g., polynucleotide) to target cells or tissues (e.g., extrahepatic cells or tissues). In certain embodiments, the Disclosure describes the use of compositions of the Disclosure in the manufacture of pharmaceuticals for use in the delivery of an active agent (e.g., polynucleotide) to target cells or tissues (e.g., extrahepatic cells or tissues). In certain embodiments, the use includes administering an effective amount of the lipid composition of the Disclosure (e.g., LNP) to a subject. In certain embodiments, the use includes administering a pharmaceutical composition containing an effective amount of the lipid composition of the Disclosure (e.g., LNP) to a subject.

[0221] In certain embodiments, the target cells or tissues are extrahepatic cells or tissues. In certain embodiments, the extrahepatic cells or tissues include one or more (e.g., all) brain cells or tissues, lung cells or tissues, bone marrow cells or tissues, spleen cells or tissues, lymph node cells or tissues, ovarian / testicular cells or tissues, muscle cells or tissues, kidney cells or tissues, pancreatic cells or tissues, and / or cardiac cells or tissues. In certain embodiments, the extrahepatic cells or tissues include one or more (e.g., all) brain cells or tissues, lung cells or tissues, and / or bone marrow cells or tissues. In certain embodiments, the bone marrow cells or tissues are located within the femur. In certain embodiments, the extrahepatic cells or tissues include brain cells or tissues. In certain embodiments, the extrahepatic cells or tissues include lung cells or tissues. In certain embodiments, the extrahepatic cells or tissues include bone marrow cells or tissues. In certain embodiments, the extrahepatic cells or tissues include spleen cells or tissues. In certain embodiments, the extrahepatic cells or tissues include lymph node cells or tissues. In certain embodiments, extrahepatic cells or tissues include ovarian or testicular cells or tissues. In certain embodiments, extrahepatic cells or tissues include muscle cells or tissues. In certain embodiments, extrahepatic cells or tissues include kidney cells or tissues. In certain embodiments, extrahepatic cells or tissues include pancreatic cells or tissues. In certain embodiments, extrahepatic cells or tissues include cardiac cells or tissues.

[0222] In certain embodiments, the Disclosure provides a method for treating a disease in a subject, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the lipid composition of the Disclosure (e.g., LNP).

[0223] In certain embodiments, the Disclosure describes a composition of the Disclosure for use in the treatment of a disease in a subject by administering a therapeutically effective amount of the composition to the subject, wherein the composition comprises a lipid composition of the Disclosure (e.g., LNP).

[0224] In certain embodiments, the Disclosure describes the use of the Compositions of the Disclosure for use in the treatment of a disease in a subject by administering a therapeutically effective amount of the Composition to the subject, wherein the Composition comprises the Lipid Compositions of the Disclosure (e.g., LNPs).

[0225] In certain embodiments, the Disclosure describes the use of a composition of the Disclosure in the manufacture of a pharmaceutical for use in the treatment of a disease in a subject by administering a therapeutically effective amount of the pharmaceutical to the subject, wherein the pharmaceutical comprises a lipid composition of the Disclosure (e.g., LNP).

[0226] In certain embodiments, the lipid composition (e.g., LNP) comprises a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof and an active agent (e.g., DNA, RNA).

[0227] In certain embodiments, the lipid composition (e.g., LNP) comprises a lipid compound selected from any one of compounds 1 to 209 or a pharmaceutically acceptable salt thereof, and encapsulates an active agent (e.g., DNA, RNA).

[0228] In certain embodiments, the lipid composition (e.g., LNP) comprises a lipid compound selected from any one of compounds 7, 8, 10, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, or 109, or a pharmaceutically acceptable salt thereof, and encapsulates an active agent (e.g., DNA, RNA).

[0229] In certain embodiments, the Disclosure provides a method for producing a therapeutic composition, the method comprising encapsulating an active agent (e.g., DNA, RNA) within lipid nanoparticles (LNPs) of the Disclosure. In certain embodiments, the Disclosure provides a method for producing a prophylactic compound, the method comprising encapsulating an active agent (e.g., DNA, RNA) within lipid nanoparticles (LNPs) of the Disclosure. In certain embodiments, the Disclosure provides a method for producing a vaccine, the method comprising encapsulating an active agent (e.g., DNA, RNA) within lipid nanoparticles (LNPs) of the Disclosure.

[0230] In certain embodiments, the lipid composition (e.g., LNP) of a therapeutic, prophylactic, or vaccine comprises a lipid compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. In certain embodiments, the active agent (e.g., DNA, RNA) is encapsulated within the lipid nanoparticles (LNP) by mixing a solution containing the active agent (e.g., DNA, RNA) with a solution / container containing lipid nanoparticles or precursor elements of lipid nanoparticles. Examples of solutions or solvents that may be used to form LNPs or encapsulate the active agent within LNPs include, but are not limited to, water, PBS, Tris buffer, NaCl, citrate buffer, acetate buffer, ethanol, chloroform, diethyl ether, cyclohexane, tetrahydrofuran, methanol, and isopropanol.

[0231] In certain embodiments, the active agent includes DNA. In certain embodiments, the active agent includes RNA. In certain embodiments, the active agent includes both DNA and RNA. In certain embodiments, the RNA includes mRNA. In certain embodiments, the RNA includes inhibitory RNA, such as siRNA.

[0232] III. Definition Unless otherwise specified, the following terms and phrases have the meanings set forth below. Definitions are not intended to be inherently limiting, but rather to provide a clear understanding of certain aspects of this disclosure.

[0233] To administer: As used herein, the term “to administer” means to provide the composition to the subject.

[0234] Alkenoxy: As used herein, the term “alkenoxy” means an alkenyl moiety linked via a divalent oxygen crosslink (e.g., -OC). 1~20 This refers to alkenyl groups. Examples of such groups include, but are not limited to, ethenoxy and propenoxy.

[0235] Alkenyl: As used herein, the term “alkenyl” refers to an unsaturated hydrocarbon chain (branched or unbranched) having one or more carbon-carbon double bonds in the chain. In certain embodiments, the alkenyl group may be optionally substituted with one or more substituents. Examples of alkenyl groups include, but are not limited to, ethirenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0236] Alkenylene: As used herein, the term "alkenylene" refers to a divalent alkenyl group. Examples of alkenylene groups include, but are not limited to, ethenylene, propenylene, butylene, pentenylene, and hexenylene.

[0237] Alkoxy: As used herein, the term “alkoxy” means an alkyl moiety linked via a divalent oxygen crosslink (e.g., ROC 1~20 This refers to a group of groups that do not contain methoxy, ethoxy, or propoxy molecules.

[0238] Alkyl: As used herein, the term “alkyl” refers to a saturated hydrocarbon chain (branched or unbranched). In certain embodiments, alkyl groups may be optionally substituted with one or more substituents (i.e., alkyl groups may be unsubstituted or substituted with one or more substituents). In certain embodiments, alkyl groups may be optionally substituted with one or more halo groups (e.g., F). Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (n-propyl, iso-propyl), butyl (n-butyl, sec-butyl, iso-butyl, tert-butyl), and pentyl (n-pentyl, isopentyl, neopentyl).

[0239] Alkylene: As used herein, the term "alkylene" refers to a divalent alkyl group. Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene (n-propylene, iso-propylene), butylene (n-butylene, sec-butylene, iso-butylene, tert-butylene), and pentylene (n-pentylene, isopentylene, neopentylene).

[0240] Alkynyl: As used herein, the term "alkynyl" refers to an unsaturated hydrocarbon chain (branched or unbranched) having one or more carbon-carbon triple bonds in the chain. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0241] Alkynylene: As used herein, the term "alkynylene" refers to a divalent alkynyl group. Examples of alkynylene groups include, but are not limited to, ethynylene, propynylene, butynylene, pentynylene, and hexynylene.

[0242] Approximately / About: As used herein, the terms “approximately” and “about” are used interchangeably and, when applied to one or more reference values, refer to values ​​within + / -10% of the stated value. In certain embodiments, unless explicitly stated or clearly evident from the context, this term refers to a range of values ​​that fall within + / -10%, + / -9%, + / -8%, + / -7%, + / -6%, + / -5%, + / -4%, + / -3%, + / -2%, + / -1%, or less than or equal to the stated reference value.

[0243] Aryl: As used herein, the term “aryl” refers to a monocyclic aromatic hydrocarbon ring or a polycyclic group comprising at least one aromatic hydrocarbon ring. In certain embodiments, the aryl group may be optionally substituted with one or more substituents. Examples of aryl groups include, but are not limited to, phenyl and naphthyl.

[0244] Arylene: The term "arylene" refers to a polyvalent (e.g., divalent or trivalent) aryl group.

[0245] Cycloalkyl: As used herein, the term “cycloalkyl” refers to a saturated monocyclic or polycyclic (e.g., bicyclic or tricyclic) hydrocarbon ring. In certain embodiments, a cycloalkyl group may be optionally substituted with one or more substituents. In certain embodiments, a cycloalkyl group may be one or more C1-C12 12 The cycloalkyl group can be optionally substituted with alkyl groups (e.g., methyl, ethyl, n-pentyl, n-butyl, n-pentyl, etc.). The number of ring atoms in the cycloalkyl ring can be specified using the "Cx~Cy cycloalkyl" nomenclature, where x and y are integers that specify the number of ring atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and adamantyl. In certain embodiments, the cycloalkyl group includes condensed cycloalkyl groups (e.g., condensed bicyclic or tricyclic groups). In certain embodiments, the cycloalkyl group includes crosslinked cycloalkyl groups (e.g., crosslinked bicyclic groups). In certain embodiments, the cycloalkyl group includes spirocycloalkyl groups (e.g., spirodicyclic groups).

[0246] Cycloalkylene: As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl group. Examples of such groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and adamantylene.

[0247] Dienyl: As used herein, the term “dienyl” refers to an unsaturated hydrocarbon chain (branched or unbranched) having one or more carbon-carbon double bonds in the chain. Examples of a dienyl group include, but are not limited to, 1,3-pentadienyl, 1,4-hexadienyl, and (2Z,5Z)undeca-2,5-dienyl.

[0248] Effective dose: As used herein, the term “effective dose” or “therapeutic effective dose” of a drug is the amount sufficient to produce a beneficial or desired outcome (e.g., a biological, medical, or clinical outcome). Therefore, the effective dose depends on the context in which it is applied (e.g., route of administration, severity of condition, biochemistry, and medical history of the subject) and can be determined by standard clinical techniques of those skilled in the art (e.g., extrapolation from dose-response curves derived from studies).

[0249] Halo: As used herein, the term "halo" refers to fluoro, chloro, bromo, and / or iodine.

[0250] Heterocyclic / Heterocyclic: As used herein, the terms “heterocyclic,” “heterocyclic,” or “heterocyclyl” refer to a saturated or unsaturated non-aromatic ring (monocyclic or bicyclic) containing one or more (e.g., 1 to 4) heteroatoms (e.g., N, O, or S). The number of ring atoms in a heterocyclic ring can be specified using the “x-y member” nomenclature, where x and y are integers specifying the number of ring atoms. For example, a 3-6 member heterocyclic group refers to a saturated or unsaturated 3-6 member ring structure containing one or more heteroatoms, e.g., nitrogen, oxygen, and sulfur. Examples of heterocyclic groups include, but are not limited to, pyrrolinyl, pyrrolidinyl, pyrazolidinyl, oxazolyl, thiazolyl, piperidinyl, piperazinyl, morpholinyl, and tetrahydropyranil.

[0251] Heterocyclylene: As used herein, the term “heterocyclylene” refers to a divalent heterocyclic group. Examples of such groups include, but are not limited to, pyrrolidinylene, pyrrolidinylene, pyrazolidinylene, oxazolylene, thiazoylene, piperidinylene, piperazylene, and morpholinylene.

[0252] Hydroxyalkyl: As used herein, the term "hydroxyalkyl" means RC 1~20This refers to the -OH group. Examples of such groups include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, and 3-hydroxypropyl.

[0253] Lipid nanoparticles: As used herein, the term “lipid nanoparticles” or “LNP” refers to particles containing multiple lipid molecules physically bound to one another by intermolecular forces. In certain embodiments, lipid nanoparticles are microspherical vesicles containing a liposome lamellar phase lipid bilayer (monolayer or multilayer). In other embodiments, lipid nanoparticles are nanospherical vesicles containing a liposome lamellar phase lipid bilayer (monolayer or multilayer).

[0254] Mole %: As used herein, the term “molar percentage” or “mol %” refers to the mole fraction of a particular element in a mixture, expressed as a percentage of the total number of moles in the mixture. A mole fraction is the number of moles of one component in a given mixture relative to the total number of moles in the mixture.

[0255] Pharmacologically acceptable: As used herein, the terms “pharmaceutically acceptable” or “therapeutably acceptable” are used to describe compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues, within the bounds of sound medical judgment, without excessive toxicity, irritation, allergic response or other problems or complications, and with a reasonable benefit / risk ratio.

[0256] pharmaceutically acceptable excipients: As used herein, the term “pharmaceutically acceptable excipient” means a component in a pharmaceutical composition that can suspend, retain, dilute, stabilize, control, encapsulate or otherwise complement a compound or composition of the present disclosure (e.g., LNP) in the pharmaceutical composition. A pharmaceutically acceptable excipient is substantially non-toxic, non-inflammatory and otherwise pharmaceutically acceptable in the subject (as defined above).

[0257] pharmaceutically acceptable salts: As used herein, the phrase “pharmaceutically acceptable salts” means salts that, within the bounds of sound medical judgment, do not have excessive toxicity, irritation, allergic reactions, etc., and are suitable for use in contact with human and lower animal tissues, and that are commensurate with a reasonable benefit / risk ratio. The compounds described herein may contain basic, acidic, or both functional groups and may be converted to pharmaceutically acceptable salts by using appropriate acids or bases as necessary. Salts may be prepared in situ during the final isolation and purification of the compounds.

[0258] Phospholipids: As used herein, the term “phospholipid” refers to a lipid comprising a phosphate moiety and one or more carbon chains, such as an unsaturated fatty acid chain. Phospholipids may contain one or more double or triple bonds (e.g., one or more unsaturated) in their carbon chains.

[0259] Prevention or Prophylaxis: As used herein, the terms “prevention” or “prophylaxis” mean to partially or completely delay the onset of a disease or condition, to partially or completely delay the onset of one or more symptoms, features or clinical manifestations of a disease or condition, to partially or completely delay the progression of a disease or condition, and / or reduce the risk of developing a pathological condition associated with a disease or condition. In certain embodiments, “prevention” or “prophylaxis” of a disease or condition may be considered a subset of the meaning of the terms “treatment” or “treatment” of a disease or condition.

[0260] Subject: As used herein, the term “Subject” means any organism to which the compositions provided herein may 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. Subjects may be seeking or needing treatment, requesting treatment, receiving or being treated, or receiving care from a trained professional for a particular disease or condition.

[0261] Identical substituents: As used herein, when two substituents are “the same,” the substituents have the same molecular formula and arrangement of bonded atoms, but their three-dimensional orientations in space may or may not be different. In certain embodiments, if each of the same substituents contains one or more double bonds, the stereochemistry of each of the one or more double bonds in one substituent is the same as the stereochemistry of the corresponding one or more double bonds in the other substituent.

[0262] Treatment: As used herein, the term “treatment” or “therapy” means to partially or completely reduce, remit, improve, alleviate, delay the onset of, inhibit the progression of, reduce the severity of, reduce the incidence of, and / or prevent one or more symptoms or characteristics of a particular disease or condition. Treatment may be administered to subjects who show no signs of the disease or condition and / or subjects who show only the initial signs of the disease or condition, for the purpose of reducing the risk of developing a disease (or further disease) associated with the disease or condition.

[0263] General Considerations In various parts of this disclosure, substituents or properties of the compounds disclosed are disclosed in groups or ranges. This disclosure includes each of the individual members or subcombinations of members of such groups and ranges, and such groups or ranges are intended to include endpoints. As a non-limiting example, if a group or range is about 1 to about 10, the group or range includes both the value about 1 and the value about 10.

[0264] Articles such as “a,” “an,” and “that” may mean one or more than one unless otherwise stated or evident from the context. Claims or descriptions containing “or” between one or more members of a group are deemed satisfied if one, two or more, or all of the group members are present in, used in, or otherwise related to a given product or process, unless otherwise stated or evident from the context. This disclosure may include embodiments in which exactly one member of a group is present in, used in, or otherwise related to a given product or process. This disclosure may include embodiments in which two or more, or all, group members are present in, used in, or otherwise related to a given product or process.

[0265] The term "contains" is intended to be open and may include, but is not required, additional elements or processes. Where the term "contains" is used herein, the terms "consist of" and "essentially consist of" are also included and disclosed.

[0266] The abbreviation "e.g." (e.g.) derives from the Latin "exempli gratia" and is used herein to indicate non-restrictive examples. Therefore, the abbreviation "e.g." (e.g.) derives from the Latin "id est" and is used herein to indicate non-restrictive paraphrasing or clarification. Therefore, the abbreviation "i.e." (e.g.) is synonymous with the term "in other words."

[0267] When a variable is provided in relation to an organic chemical structure as having a range of numbers, it is understood that the variable is an integer value within that range, including the endpoint. For example, "n is 0 to 3" means that n is 0, 1, 2, or 3.

[0268] Any embodiment of the present disclosure that falls within the scope of the prior art may be expressly excluded from any one or more claims. Any embodiment of the agents, methods and / or compositions of the present disclosure may be excluded from any one or more claims for any reason, whether or not it relates to the existence of the prior art.

[0269] Publications, patent applications, patents, and other references described herein are incorporated by reference, and in the event of any conflict between this specification and any other specification, this specification shall prevail.

[0270] The section headings, materials, methods, and examples are illustrative and not intended to be limiting. [Examples]

[0271] Example 1 - Preparation and analysis of lipid nanoparticles (LNPs) a. LNP preparations Formulations containing the lipid compounds described herein were prepared. Four common formulations, as shown in Table 2, were used.

[0272] [Table 71]

[0273] LNP formulations were prepared using the lipid compounds listed in Table 1: compounds 1-12, 14, 16, 18, and 20. The preparation results are shown in Table 3 (PDI = polydispersity index, N / P = molar ratio of ionic nitrogen to phosphate group, EE% = encapsulation efficiency (measured using a fluorescence plate-based assay)).

[0274] [Table 72]

[0275] [Table 73]

[0276] [Table 74]

[0277] [Table 75]

[0278] b.LNP preparation Lipids and active ingredients (e.g., RNA, DNA, etc.) were assembled into LNPs using microfluidic mixing. The ethanol phase was prepared by solubilizing ionized lipids (i.e., lipid compounds), phospholipids (e.g., DSPC), cholesterol, and PEG lipids in ethanol at predetermined molar ratios shown in Table 2. The aqueous phase was prepared by diluting nucleic acid cargo (e.g., firefly luciferase mRNA) with acidic buffer (pH 4.0, citrate buffer, 50 mM). The two phases were mixed at predetermined flow rates (e.g., 9 mL / min for the aqueous phase and 3 mL / min for the ethanol phase) using a tip equipped with a microfluidic mixing structure (e.g., toroidal mixer). The resulting LNPs, containing lipids and encapsulating nucleic acid cargo, were dialyzed for 2 hours in PBS at pH 7.4 at room temperature using a 20 kDa MWCO dialysis membrane. Then, fresh PBS was supplied and dialysis was continued overnight at 2–8°C. After buffer exchange was completed, LNPs were selectively concentrated using an ultracentrifuge equipped with a 10-100 kDa MWCO membrane.

[0279] c. Plate-based fluorescence (RiboGreen) assay Nucleic acid cargo in LNPs was quantified against a nucleic acid standard curve using the RiboGreen assay. LNPs containing nucleic acid cargo were separately mixed with 1x Tris-EDTA (TE) and 2% Triton in TE to achieve the required dilution for the assay. The samples were then heated at 30°C and vortex-mixed at 300 rpm. The samples were seeded into 96-well plates, and RiboGreen reagent was added to each well. After incubation in the dark, fluorescence from the wells was measured and plotted against the standard curve. The total cargo concentration was quantified using the nucleic acid concentration obtained from the Triton-treated samples. Unencapsulated (free) cargo was determined using only the fluorescence values ​​of the TE-diluted samples, and the encapsulation efficiency (EE%) was determined using its ratio.

[0280] d. Dynamic light scattering Based on the process steps and LNP concentration, LNP was diluted with PBS (pH 7.4) to the specified dilution range. After mixing, particle size and PDI were measured using a dynamic light scattering instrument.

[0281] Example 2 - Stability Test LNPs prepared from selected lipid compounds were tested for stability under conditions of 2–8°C. Compounds 1–7 and 10 (N / P:6) were tested with formulation F1 for 4 weeks. Particle size and PDI results are shown in Figures 1A and 1B.

[0282] Compounds 2, 3, 5, and 7 were also tested in formulation F3 over a two-week period. Particle size and PDI results are shown in Figures 1C and 1D.

[0283] Example 3 - In vivo activity in mice a. General protocols In vivo distribution studies in small rodents were conducted in mice (e.g., C57BL / 6J, Balb-c, CD-1, etc.). To evaluate the in vivo distribution and tissue activity of LNPs after systemic administration, C57BL / 6J mice were administered firefly luciferase mRNA at different dose levels (0.25–3 mg / kg) via single intravenous injection through the tail vein of LNP preparations (individually prepared or pooled) in PBS. At predetermined time points (4–6 hours post-injection), the animals were anesthetized with isoflurane and subjected to bioluminescence imaging sessions using an in vivo imaging system (IVIS). All animals were administered 15 mg / mL of D-luciferin by subcutaneous (SC) injection at a dose of 0.2 mL / animal. The animals were shaved of their abdominal hair using an animal trimmer. The animals were then positioned so that their shaved abdomens faced the IVIS camera. Whole-body imaging sessions were performed 10–15 minutes after D-luciferin administration.

[0284] Ex vivo imaging sessions were also conducted to investigate the detailed organ distribution and activity of LNP preparations. All animals received SC D-luciferin, were then euthanized by an overdose of isoflurane, and subsequently perfused with saline. After perfusion, organs were collected and subjected to IVIS imaging for luminescence within 10-15 minutes after D-luciferin injection.

[0285] For lung delivery, LNP preparations were administered to CD-1 mice by single intratracheal injection under isoflurane anesthesia. Animals were also administered 15 mg / mL of D-luciferin subcutaneously (SC) at a dose of 0.2 mL / animal. A whole-body imaging session was performed within 10–15 minutes after D-luciferin injection. The animals were then perfused with saline, and both lungs and tracheas were collected and imaged for luminescence.

[0286] All IVIS images were processed using computer software to identify regions of interest in individual organs in order to detect total flux (p / s) values ​​as a quantification of luminescence. The total flux values ​​were then graphed to evaluate in vivo activity in each collected organ.

[0287] b. General activity tests LNPs prepared from selected lipid compounds were tested in mice for in vivo mRNA delivery activity. Following the preparation of LNPs using the indicated lipid compounds according to each of the four formulations described in Example 2, the four LNP formulations were pooled together. An additional pool was created by combining sets of four LNPs corresponding to the four different lipid compounds shown, resulting in 16 different compositions. The formulation characteristics of the resulting LNPs are shown in Table 4.

[0288] [Table 76]

[0289] Each LNP compound pool was injected (IV) into mice (1 mg / kg), and PBS was used as a control. Mouse organ imaging results showing the total flux [p / s] after 6 hours are shown in Figures 2A, 2B, 2C, and 2D.

[0290] c. Brain activation The LNP compound pools of compounds 1, 4, 6, and 10 were administered locally (ICV) to mouse brains (1 μg / animal), and PBS was used as a control. Figure 3 shows the mouse brain imaging results, which indicate the total flux [p / s].

[0291] d. Lung activity LNP compound pools of compounds 2, 3, 5, and 7 were administered locally (intratracheally) to mouse lungs (7 μg RNA / animal), and PBS was used as a control. Mouse lung imaging results showing total flux [p / s] are shown in Figure 4A (in vivo imaging) and Figure 4B (ex vivo imaging).

[0292] LNP compound pools of compounds 8, 9, 11, and 20 were administered locally (intratracheally) to mouse lungs (7 μg RNA / animal), and PBS was used as a control. Mouse lung imaging results showing total flux [p / s] are shown in Figure 4C.

[0293] The LNP compound pools of compounds 12, 14, 16, and 18 were administered locally (intratracheally) to mouse lungs (7 μg RNA / animal), and PBS was used as a control. The results of mouse lung imaging showing total flux [p / s] are shown in Figure 4D.

[0294] Example 4 - Formulation F2 Test a. Formulation F2 study design LNPs incorporating lipid compounds 7, 8, 10, 14, and 18 were prepared using formulation F2 of Example 1. For comparison, an LNP formulation incorporating MC3 (Cayman Chemical) as the lipid compound was also prepared using formulation F2. PBS was used as a control. The formulation characteristics are shown in Table 5.

[0295] [Table 77]

[0296] b. Activity of the general formulation F2 Each LNP formulation was injected (IV) into mice (0.3 mg / kg), and PBS was used as a control. Imaging results were collected according to the protocol of Example 4. Figure 5A shows mouse organ imaging results showing the total flux [p / s] after 6 hours. Figure 5B shows mouse organ imaging results normalized to MC3 activity.

[0297] c. Lung tests of formulation F2 Each LNP preparation was administered locally (intratracheally) to the lungs of mice (7 μg RNA / animal), MCS was administered as a comparison, and PBS was used as a control. Imaging results were collected according to the protocol of Example 4. Imaging results of mouse lungs and tracheas showing total flux [p / s] are shown in Figure 6A (live animal imaging) and Figure 6B (ex vivo imaging).

[0298] Example 5 - In vivo activity screening in mice A biodistribution study in mice was conducted in small rodents according to the general test protocol of Example 3(a). LNPs incorporating lipid compounds were prepared using formulation F2, and for comparison, MC3 (Cayman Chemical) was used as the lipid compound prepared using formulation F2. LNP pools (containing firefly luciferase mRNA) were injected into mice at various doses, and then organ-average total flux (p / s) results (measuring luciferase activity) were collected according to Example 3(a).

[0299] Table 6 shows the LNP dose, LNP pool composition, and organ-average total flux results in multiple mouse organs. LNPs containing MC3 were used as controls and for inter-mouse comparisons. Doses are expressed in mg / kg units, and LC represents the lipid compound according to Table 1.

[0300] [Table 78]

[0301] [Table 79]

[0302] [Table 80]

[0303] Additional screening results for compounds 7, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, and 109 in the spleen, femur, and muscle are shown in Figures 7A, 7B, and 7C. MC3 was used as a control for comparison.

[0304] Example 6 - In vivo activity screening in non-human primates In vivo distribution studies of non-human primates (NHPs) were conducted in 4-6 year old cynomolgus monkeys (e.g., Macaca fascicularis). Two days and one day prior to administration of the test substance, the animals were pre-administered dexamethasone, famotidine, and diphenhydramine (1, 0.5, and 5 mg / kg, respectively). LNP formulations (prepared with formulation F2 of Example 1) were prepared using ionized lipids so that each LNP formulation contained one or more mRNAs with a predetermined unique barcode sequence as cargo. LNPs in PBS, identified by the unique barcode mRNA cargo, were then pooled together in predetermined ratios. To evaluate the biodistribution of LNPs to tissues after systemic administration, NHPs received doses of LNP administration material (individually or pooled) in PBS at different total dose levels (0.5–1 mg / kg) via a single intravenous infusion over 30 minutes via a peripheral vein (radial vein, saphenous vein, or other available vein).

[0305] At a predetermined time (4–6 hours after administration of the pooled test substance), ketamine (10 mg / kg) and Euthasol® (0.25 mL / kg) were intravenously administered, and the animals were sacrificed by bleeding from the femoral artery. Subsequently, all animals underwent systemic perfusion with PBS before tissue sampling. 20–25 mg samples for in vivo distribution analysis were collected from all treatment groups and placed in sample tubes containing RNALater® solution. All tissue samples were stored at room temperature for 24 hours, after which the supernatant was removed and the samples were frozen.

[0306] To determine the unique LNPs distributed within the tissues, samples were homogenized using a tissue homogenizer (e.g., TissueLyser). Total RNA was extracted from the tissue homogenates using standard methods. Then, to ensure accurate counting of RNA molecules, a normalized amount of RNA was sequenced using a targeted RNA sequencing method with unique molecular identifiers (UMIs). For each tissue, the obtained RNA molecule counts were normalized by the RNA counts of a pooled test material. The results were analyzed using our in-house bioinformatics pipeline.

[0307] Table 7 shows the LNP pool composition and UMI counts in multiple NHP organs. LNPs containing MC3 were used as controls and for inter-mouse comparisons. Doses are in mg / kg units, and LC is the lipid compound according to Table 1.

[0308] [Table 81]

[0309] [Table 82]

[0310] [Table 83]

[0311] The screening results for NHP group 1 (spleen, femur, muscle) are shown in Figures 8A, 8B, and 8C. The screening results for NHP group 2 (spleen) are shown in Figure 9. The screening results for NHP group 3 (femur) are shown in Figure 10. The screening results for NHP group 4 (muscle) are shown in Figure 11.

[0312] Example 7 - General materials and methods for synthesis All temperatures are in degrees Celsius and are uncorrected. Reagent-grade chemicals and anhydrous solvents were purchased commercially and used without further purification unless otherwise specified. Product names were determined using the naming software included in the Biovia electronic laboratory notebook. Silica gel chromatography was performed using a Teledyne Isco instrument with a pre-packaged disposable SiO2 stationary phase column (eluent flow rate range 15–200 mL / min, UV detection (254 and 280 nm)). Reverse-phase purification was performed using a C18 column and UV detection (214 and 254 nm). Chemical shifts are reported in ppm. 1 In 1H NMR, the solvent peaks appearing at 7.26 ppm for CDCl3, 2.50 ppm for DMSO d6, and 3.31 ppm for CD3OD are used as reference points.

[0313] Terms and abbreviations: 4-PPY 4-pyrrolidine-1-ylpyridine, Ac acetyl, AQ water-based, Bn benzyl, DCM dichloromethane, DIPEA N,N-diisopropylethylamine, DMAP 4-dimethylaminopyridine, EDCI 1-Ethyl 3(3-dimethylaminopropyl)carbodiimide hydrochloride, Et ethyl,  Acetyl acetate, h time, HCl hydrochloride, HPLC (High-Performance Liquid Chromatography), LCMS Liquid Chromatography Mass Spectrometry, mCPBA 3-chloroperbenzoic acid, Me methyl, min minutes, NaHCO3 (sodium bicarbonate) Na2SO4 sodium sulfate, PPTS Pyridinium-p-toluenesulfonate, Py pyridine, TEA triethylamine, TEMPO 1-Oxidanyl-2,2,6,6-tetramethylpiperidine, NH4Cl ammonium chloride, NMR nuclear magnetic resonance, p-TsOH-H2O para-toluenesulfonic acid monohydrate, sat. saturation, TBS tert-butyldimethylsilyl, THF (Tetrahydrofuran).

[0314] Example 8 - Compound 1: 2-((oleoyloxy)methyl)-2-(((4-(((2-(pyrrolidine-1-yl)ethyl)carbamoyl)oxy)decanoyl)oxy)methyl)propane-1,3-diyldioleate Step 1: [ka] NaOH (24.67 g, 616.75 mmol, 1.05 equivalent) was slowly added to a solution of 5-hexyltetrahydrofuran-2-one (100 g, 587.38 mmol, 1 equivalent) in H2O (500 mL). This mixture was stirred at 100 °C for 12 hours under N2. The reaction mixture was concentrated under reduced pressure to remove the solvent. Compound 4-hydroxydecanoyloxysodium (90 g, crude) was obtained as a white solid and was used directly in the next step.

[0315] Step 2: [ka] Bromomethylbenzene (8.13 g, 47.56 mmol, 5.65 mL, 1 equivalent) was added dropwise to a solution of 4-hydroxydecanoyl oxysodium (10 g, 47.56 mmol, 1 equivalent) in DMSO (100 mL). This mixture was stirred under N2 at 25°C for 5 minutes. The reaction mixture was diluted with saturated NaCl (100 mL) and extracted with siRNA (200 mL) (100 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound benzyl 4-hydroxydecanoate (12 g, crude) was obtained as a pale yellow oil, which was immediately used in the next step.

[0316] Step 3: [ka] To a solution of benzyl 4-hydroxydecanoate (12 g, 43.11 mmol, 1 equivalent) in DCM (120 mL), (4-nitrophenyl)carbonochloride (17.38 g, 86.21 mmol, 2 equivalents) and Py. (6.82 g, 86.21 mmol, 6.96 mL, 2 equivalents) were slowly added at 0°C. This mixture was stirred under N2 at 25°C for 1 hour. The reaction mixture was diluted with petroleum ether (100 mL), filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound benzyl 4-(4-nitrophenoxy)carbonyl oxydecanoate (10 g, 22.55 mmol, yield 52.31%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 8.30-8.25(m,2H)7.37-7.27(m,7H)5.14(s,2H)4.91-4.85(m,1H)2.53-2.50(t,J=7.6Hz,2H)2.16-2.07( m,1H)2.05-1.97(m,1H)1.77-1.72(m,1H)1.67-1.60(m,1H)1.43-1.30(m,8H)0.91-0.88(t,J=6.0Hz,3H).

[0317] Step 4: [ka] To a solution of benzyl 4-(4-nitrophenoxy)carbonyl oxydecanoate (10 g, 22.55 mmol, 1 equivalent) in DCM (100 mL), 2-pyrrolidine-1-ylethaneamine (7.72 g, 67.65 mmol, 3 equivalents), DMAP (550.94 mg, 4.51 mmol, 0.2 equivalents), and DIPEA (8.74 g, 67.65 mmol, 11.78 mL, 3 equivalents) were added. This mixture was stirred under N2 at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 200 mL (100 mL x 2) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). The compound benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoate (7g, 16.72 mmol, yield 74.17%) was obtained as a colorless oil.

[0318] Step 5: [ka] Benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoate (7g, 16.72 mmol, 1 equivalent) was added to a suspension of Pd / C (2g, 1.88 mmol, 10% purity, 0.112 equivalents) in THF (140 mL). The mixture was stirred at 25°C for 12 hours under H2 (15 Psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 50 / 1~3 / 1). Compound 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (3g, 9.13 mmol, yield 54.62%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 10.23(s,1H)6.27(s,1H)4.77-4.71(m,1H)3.68-3.60(m,1H)3.19-3.14(m,1H)3.08-2.95(m,5H)2.82-2.77(m,1H)2.37-2.21(m,2H) )2.06-2.00(m,1H)1.96(s,4H)1.84-1.74(m,1H)1.64-1.58(m,1H)1.52-1.45(m,1H)1.30-1.26(m,8H)0.88-0.85(t,J=6.8Hz,3H).

[0319] Step 6: [ka] A mixture of [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (8g, 45.40 mmol, 0.45 equivalents), (Z)-octadeca-9-enoic acid (28.50g, 100.89 mmol, 28.50mL, 1 equivalent), EDCI (23.21g, 121.07 mmol, 1.2 equivalents), DIPEA (32.60g, 252.22 mmol, 43.93mL, 2.5 equivalents), and DMAP (1.23g, 10.09 mmol, 0.1 equivalent) in 285mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200mL) and extracted with 600mL (200mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). The compound [2,2-dimethyl-5-[[(Z)-octadeca-9-enoyl]oxymethyl]-1,3-dioxan-5-yl]methyl(Z)-octadeca-9-enoate (20 g, 28.36 mmol, yield 56.23%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.37-5.34(m,4H)4.11(s,4H)3.75(s,4H)2.34-2.30(t,J=7.6Hz,4H)2.05-1.99(m ,8H)1.63-1.60(m,4H)1.43(s,6H)1.31-1.27(m,40H)0.91-0.87(t,J=6.8Hz,6H).

[0320] Step 7: [ka] [2,2-dimethyl-5-[[(Z)-octadeca-9-enoyl]oxymethyl]-1,3-dioxan-5-yl]methyl(Z)-octadeca-9-enoate (20 g, 28.36 mmol, 1 equivalent) was slowly added to a solution of THF (200 mL) with HCl (3 M, 10.40 mL, 1.1 equivalents) at 0°C. The mixture was then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was adjusted to pH=7 at 0°C with saturated Na2CO3 and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1~1 / 1). The compound [2,2-bis(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (10 g, 15.04 mmol, yield 53.01%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.38-5.34(m,4H)4.15(s,4H)3.58(s,4H)2.73(s,2H)2.38-2.34(t,J=7.2Hz,4H)2 .07-2.00(m,8H)1.65-1.60(m,4H)1.31-1.25(m,40H)0.91-0.87(t,J=6.8Hz,6H).

[0321] Step 8: [ka] To a 40 mL solution of DCM (40 mL) containing [2,2-bis(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (4 g, 6.01 mmol, 1 equivalent), EDCI (1.38 g, 7.22 mmol, 1.2 equivalents), DMAP (73.48 mg, 601.47 μmol, 0.1 equivalent), and DIPEA (1.94 g, 15.04 mmol, 2.62 mL, 2.5 equivalents), oleic acid (1.70 g, 6.01 mmol, 1.70 mL, 1 equivalent) from DCM (40 mL) was slowly added. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound [2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxy-2-[[(Z)-octadeca-9-enoyl]oxymethyl]propyl](Z)-octadeca-9-enoate (1.5 g, 1.61 mmol, yield 26.83%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.40-5.31(m,6H)4.12(s,6H)3.50-3.49(d,J=6.8Hz,2H)2.53-2.49(t,J=7.2Hz,1H)2.35-2.31(t ,J=7.2Hz,6H)2.07-2.00(m,12H)1.64-1.58(m,6H)1.31-1.28(m,60H)0.90-0.87(t,J=6.8Hz,9H).

[0322] Step 9: [ka] [2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxy-2-[[(Z)-octadeca-9-enoyl]oxymethyl]propyl](Z)-octadeca-9-enoate (1.5 g, 1.61 mmol, 1 equivalent) was slowly added to a mixture of 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (477.04 mg, 1.45 mmol, 0.9 equivalents), EDCI (371.24 mg, 1.94 mmol, 1.2 equivalents), DMAP (19.72 mg, 161.38 μmol, 0.1 equivalent), and DIPEA (521.43 mg, 4.03 mmol, 702.74 μL, 2.5 equivalents) in 15 mL of DCM. The reaction mixture was diluted with water (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). Compound [2,2-bis[[(Z)-octadeca-9-enoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](Z)-octadeca-9-enoate (104 mg, 83.88 μmol, yield 5.20%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.80(s,1H)5.39-5.32(m,6H)4.75-4.71(m,1H)4.17-4.08(m,8H)3.5 2-3.50(m,2H)3.20-2.94(m,4H)2.39-2.35(m,2H)2.33-2.29(t,J=7. 6Hz,6H)2.02-1.99(m,14H)1.94-1.86(m,2H)1.83-1.75(m,2H)1.61-1.58(m,8H)1.49-1.42(m,2H)1.30-1.27(m,68H)0.90-0.87(m,12H).

[0323] Example 9 - Compound 2: 2-((((9Z,12Z)-Octadeca-9,12-dienoyl)oxy)methyl)-2-(((4-(((2-(pyrroridine-1-yl)ethyl)carbamoyl)oxy)decanoyl)oxy)methyl)propane-1,3-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) Step 1: [ka] 2,2-Bis(hydroxymethyl)propane-1,3-diol (50 g, 367.25 mmol, 1 equivalent) was dissolved in 500 mL of DMF in a 1000 mL round-bottom flask under N2 conditions. This mixture was stirred at 90°C for 1 hour under N2 conditions. Then, 2,2-dimethoxypropane (38.25 g, 367.25 mmol, 45.00 mL, 1 equivalent) and 4-methylbenzenesulfonic acid hydrate (698.58 mg, 3.67 mmol, 0.01 equivalent) were added to the reaction mixture at 70°C under N2 conditions. This mixture was stirred at 25°C for 12 hours under N2 conditions. The reaction mixture was quenched using 10 mL of TEA and stirred at 25°C for 1 hour under N2 conditions. The mixture was then concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (28 g, 158.90 mmol, yield 43.27%) was obtained as a white solid.

[0324] Step 2: [ka] [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (8.47 g, 48.04 mmol, 1 equivalent) and (9Z,12Z)-octadeca-9,12-dienoic acid (28.29 g, 100.89 mmol, 28.29 mL, 2.1 equivalents) were mixed in DCM (280 mL) and EDCI (11.05 g, 57.65 mmol, 1.2 equivalents), DMAP (586.93 mg, 4.80 mmol, 0.1 equivalent), and DIPEA (15.52 g, 120.11 mmol, 20.92 mL, 2.5 equivalents) were added. This mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with ethyl acetate 1500 mL (500 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). Compound [2,2-dimethyl-5-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-1,3-dioxan-5-yl]methyl(9Z,12Z)-octadeca-9,12-dienoate (15 g, 21.40 mmol, yield 44.54%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.79-11.42(m,1H),5.40-5.36(m,4H),2.80-2.77(t,J=6.32Hz,2H),2.38-2.40(t,J=7.52Hz,2H) ,2.08-2.04(q,J=6.6Hz,4H),1.66-1.61(m,2H),1.37-1.31(m,14H),0.92-0.88(t,J=6.72Hz,3H).

[0325] Step 3: [ka] [2,2-dimethyl-5-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-1,3-dioxan-5-yl]methyl(9Z,12Z)-octadeca-9,12-dienoate (15 g, 21.40 mmol, 1 equivalent) was dissolved in THF (150 mL) and HCl (3 M, 7.85 mL, 1.1 equivalents) was added. This mixture was stirred at 25 °C for 7 hours. The reaction mixture was diluted with 200 mL of H₂O and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1~1 / 1). The compound [2,2-bis(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (8g, 12.10 mmol, yield 56.57%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.33-5.26(m,8H)4.07(s,4H)3.50(s,4H)2.72-2.69(t,J=6.8Hz,4H)2.30-2.26(t,J=7.2Hz,4 H)2.01-1.96(m,8H)1.58-1.54(t,J=7.2Hz,4H)1.32-1.19(m,30H)0.84-0.80(t,J=6.8Hz,6H).

[0326] Step 4: [ka] A mixture of (9Z,12Z)-octadeca-9,12-dienoic acid (424.27 mg, 1.51 mmol, 424.27 μL, 0.5 equivalents) and [2,2-bis(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (2 g, 3.03 mmol, 1 equivalent) in DCM (204 mL) was mixed with DIPEA (977.63 mg, 7.56 mmol, 1.32 mL, 2.5 equivalents), EDCI (696.04 mg, 3.63 mmol, 1.2 equivalents), and DMAP (36.96 mg, 302.57 μmol, 0.1 equivalent). This mixture was incubated at 25°C for 12 hours in an N2 atmosphere. The mixture was stirred under gaseous conditions. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL of ethyl acetate (20 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). The compound [2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxy-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]propyl](9Z,12Z)-octadeca-9,12-dienoate (700 mg, 758.04 μmol, yield 25.05%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.40-5.30(m,12H)4.12(s,6H)3.50-3.49(d,J=6.8Hz,2H)2.79-2.76(t,J=6.4Hz,6H)2.54-2.50(t,J=7.2Hz,1H )2.35-2.31(t,J=7.6Hz,6H)2.08-2.03(m,12H)1.64-1.60(m,6H)1.38-1.28(m,42H)0.91-0.88(t,J=6.8Hz,9H).

[0327] Step 5: [ka] [2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxy-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]propyl](9Z,12Z)-octadeca-9,12-dienoate (700 mg, 758.04 μmol, 1 equivalent) and 4-(2-pyrrolidine-1-ylethylcal A mixture of bamoyl(oxy)decanoic acid (497.95 mg, 1.52 mmol, 2 equivalents) in DCM (7 mL) was mixed with DMAP (27.78 mg, 227.41 μmol, 0.3 equivalents), DIPEA (244.93 mg, 1.90 mmol, 330.09 μL, 2.5 equivalents), and EDCI (174.38 mg, 909.65 μmol, 1.2 equivalents). This mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with 30 mL (10 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). The compound [2,2-bis[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](9Z,12Z)-octadeca-9,12-dienoate (110 mg, 88.24 μmol, yield 11.64%, purity 98.98%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.61(s,1H)5.42-5.30(m,12H)4.75(s,1H)4.17-4.09(m,8H)3.46(s,2H)2.94-2.85(m,2H)2.79-2.76(t,J=6.8Hz,6H)2.39-2.35(m,2H)2 .33-2.29(t,J=7.6Hz,6H)2.08-2.03(m,12H)1.96-1.89(m,4H)1.87-1.74(m,2H)1.62-1.59(m,8H)1.49-1.27(m,54H)0.94-0.86(m,12H).

[0328] Example 10 - Compound 3: [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[(Z)-octadeca-9-enoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](Z)-octadeca-9-enoate Step 1: [ka] [2,2-bis(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (3g, 4.51 mmol, 1 equivalent), EDCI (1.04g, 5.41 mmol, 1.2 equivalents), DIPEA (1.46g, 11.28 mmol, 1.96 mL, 2.5 equivalents), and DMAP (55.11 mg, 451.10 μmol, 0.1 equivalent) were mixed in 30 mL of DCM, to which 2-(1-adamantyl)acetic acid (525.81 mg, 2.71 mmol, 0.6 equivalents) in 30 mL of DCM was slowly added. The reaction mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with water (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). The compound [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (1.5 g, 1.78 mmol, yield 39.52%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.40-5.31(m,4H)4.12-4.09(m,6H)3.52-3.50(d,J=6.8Hz,2H)2.54-2.50(t,J=7.2Hz,1H)2.35-2.31(t,J= 7.6Hz,4H)2.10(s,2H)2.04-1.98(m,9H)1.73-1.57(m,16H)1.31-1.28(m,42H)0.91-0.87(t,J=6.8Hz,6H).

[0329] Step 2: [ka] [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (1 g, 1.19 mmol, 1 equivalent) and 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (351.37 mg, 1.07 mmol, 0.9 equivalents) were mixed in 10 mL of DCM with EDCI (273.44 mg, 1.43 mmol, 1.2 equivalents), DMAP (43.56 mg, 356.60 μmol, 0.3 equivalents), and DIPEA (384.06 mg, 2.97 mmol, 517.60 μL, 2.5 equivalents). The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The compound [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[(Z)-octadeca-9-enoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](Z)-octadeca-9-enoate (105 mg, 91.17 μmol, yield 7.67%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.39-5.31(m,4H)4.75(s,1H)4.16-4.09(m,8H)3.39(s,2H)2.86-2.63(m,4H)2.40-2.35(m,2H)2.33-2.29(t,J=7.6Hz,4H)2.08(s,2H)2.0 2-1.97(m,10H)1.89(s,4H)1.82-1.77(m,2H)1.72-1.69(m,4H)1.63- 1.58(m,14H)1.49-1.42(m,2H)1.30-1.27(m,50H)0.90-0.87(m,9H).

[0330] Example 11 - Compound 4: [2-(4,4-dioctoxybutanoyloxymethyl)-2-[[(Z)-octadeca-9-enoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](Z)-octadeca-9-enoate Step 1: [ka] A mixture of 4,4-dimethoxybutanenitrile (30 g, 232.28 mmol, 1 equivalent), octan-1-ol (90.75 g, 696.83 mmol, 110.13 mL, 3 equivalents), and PPTS (145.93 g, 580.69 mmol, 2.5 equivalents) in toluene (300 ml) was degassed, purged three times with N2, and then stirred at 110°C for 36 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 50 / 1). Compound 4,4-dioctoxybutanenitrile (40 g, 122.88 mmol, yield 52.90%) was obtained as a colorless oil.

[0331] Step 2: [ka] 4,4-Dioctoxybutanenitrile (40 g, 122.88 mmol, 1 equivalent) was slowly added to a solution of EtOH (100 mL) and H2O (100 mL) with NaOH (24.57 g, 614.39 mmol, 5 equivalents). The reaction mixture was stirred at 110 °C for 12 hours under an N2 atmosphere. The reaction mixture was adjusted to pH=5 using 1 M HCl and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). Compound 4,4-Dioctoxybutanoic acid (30 g, 87.08 mmol, yield 70.86%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 4.54-4.51(t,J=5.6Hz,1H)3.60-3.56(m,2H)3.45-3.40(m,2H)2.47-2.44(t,J=7.6Hz, 2H)1.97-1.92(m,2H)1.58-1.55(m,4H)1.32-1.28(m,20H)0.90-0.87(t,J=6.8Hz,6H).

[0332] Step 3: [ka] [2,2-Bis(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (3g, 4.51 mmol, 1 equivalent), EDCI (1.04g, 5.41 mmol, 1.2 equivalents), DMAP (55.11mg, 451.10 μmol, 0.1 equivalent), and DIPEA (1.46g, 11.28 mmol, 1.96 mL, 2.5 equivalents) were dissolved in 30 mL of DCM, to which 4,4-dioctoxybutanoic acid (1.55g, 4.51 mmol, 1 equivalent) from 30 mL of DCM was slowly added. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). The compound [2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (1.5 g, 1.51 mmol, yield 33.54%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.39-5.30(m,4H)4.50-4.47(t,J=5.6Hz,1H)4.11(s,6H)3.59-3.50(m,4H)3.43-3.38(m,2H)2.61-2.57(t,J=6.8Hz,1H)2.44-2.4 0(t,J=7.6Hz,2H)2.34-2.31(t,J=7.6Hz,4H)2.02-1.91(m,10H)1.63-1.54(m,8H)1.31-1.28(m,60H)0.90-0.87(t,J=6.8Hz,12H).

[0333] Step 4: [ka] [2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[(Z)-octadeca-9-enoyl]oxypropyl](Z)-octadeca-9-enoate (1.5g, 1.51 mmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (447.18mg, 1.36 mmol, 0.9 equivalent) A mixture of EDCI (348.00 mg, 1.82 mmol, 1.2 equivalents), DMAP (55.44 mg, 453.84 μmol, 0.3 equivalents), and DIPEA (488.79 mg, 3.78 mmol, 658.75 μL, 2.5 equivalents) in 20 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (50 ml) and extracted with 150 mL (50 ml x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The compound [2-(4,4-dioctoxybutanoyloxymethyl)-2-[[(Z)-octadeca-9-enoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](Z)-octadeca-9-enoate (110 mg, 84.49 μmol, yield 5.58%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.40-5.31(m,4H)4.76(s,1H)4.49-4.47(t,J=5.2Hz,1H)4.15-4.11(m,8H)3.59-3.53(m,2H)3.43-3.38(m,4H)2 .69(s,4H)2.44-2.26(m,8H)2.02-1.99(m,8H)1.94-1.73(m,8H)1.68-1.05(m,84H)0.90-0.87(t,J=6.8Hz,15H)

[0334] Example 12 - Compound 5: [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](9Z,12Z)-octadeca-9,12-dienoate Step 1: [ka] A mixture of [2,2-bis(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (3g, 4.54 mmol, 1 equivalent), 2-(1-adamantyl)acetic acid (529.02 mg, 2.72 mmol, 0.6 equivalents), DMAP (55.45 mg, 453.86 μmol, 0.1 equivalent), EDCI (1.04 g, 5.45 mmol, 1.2 equivalents), and DIPEA (1.47 g, 11.35 mmol, 1.98 mL, 2.5 equivalents) in a 30 mL DCM was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). The compound [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (800 mg, 955.50 μmol, yield 21.05%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 7.77(d,J=5.4Hz,1H),3.97-3.87(m,1H),3.81-3.68(m,4H),3.48(s,3H),2.87-2.69 (m,6H),2.40-2.37(t,J=6.6Hz,2H),1.99-1.94(m,2H),1.20-1.17(t,J=7.2Hz,6H).

[0335] Step 2: [ka] [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (800 mg, 955.50 μmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (627.66 mg, A mixture of 1.91 mmol, 2 equivalents of 1.91 mmol, 2 equivalents of DMAP (35.02 mg, 286.65 μmol, 0.3 equivalents), DIPEA (308.73 mg, 2.39 mmol, 416.08 μL, 2.5 equivalents), and EDCI (219.81 mg, 1.15 mmol, 1.2 equivalents) in 8 mL of DCM was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). The compound [2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](9Z,12Z)-octadeca-9,12-dienoate (140 mg, 121.98 μmol, yield 12.77%) was obtained as a colorless oil. 1 ¹H NMR (400 MHz, CDCl3) δ ppm 5.54-5.24(m,8H),5.18(d,J=1.0Hz,1H),4.75(s,1H),4.16-4.03(m,8H),3.39 -3.21(m,2H),2.79-2.76(t,J=6.4Hz,4H),2.69-2.44(m,6H),2.40-2.28(m,6H ),2.10-2.01(m,10H),2.00(m,1H),1.97(s,2H),1.79(s,5H),1.72(s,1H),1.6 9(s,2H),1.65-1.54(m,14H),1.42-1.21(m,38H),0.91-0.87(q,J=6.6Hz,9H).

[0336] Example 13 - Compound 6: [2-(4,4-dioctoxybutanoyloxymethyl)-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](9Z,12Z)-octadeca-9,12-dienoate Step 1: [ka] A mixture of [2,2-bis(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (3g, 4.54 mmol, 1 equivalent), 4,4-dioctoxybutanoic acid (938.19 mg, 2.72 mmol, 0.6 equivalents), EDCI (1.04 g, 5.45 mmol, 1.2 equivalents), DMAP (55.45 mg, 453.86 μmol, 0.1 equivalent), and DIPEA (1.47 g, 11.35 mmol, 1.98 mL, 2.5 equivalents) in a 60 mL DCM was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (300 mL) and extracted with 900 mL (300 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). The compound [2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,12Z)-octadeca-9,12-dienoate (900 mg, 911.38 μmol, yield 20.08%) was obtained as a colorless oil. 11H NMR (400 MHz, CDCl3) δ ppm 5.41 - 5.32 (m, 8H), 4.50 - 4.47 (t, J = 5.2 Hz, 1H), 4.12 (s, 6H), 3.58 - 3.50 (m, 4H), 3.44 - 3.38 (m, 2H), 2.80 - 2.76 (t, J = 6.4 Hz, 4H), 2.60 - 2.56 (t, J = 6.8 Hz, 1H), 2.44 - 2.40 (t, J = 7.2 Hz, 2H), 2.35 - 2.31 (t, J = 7.6 Hz, 4H), 2.08 - 2.03 (q, J = 6.8 Hz, J = 13.6 Hz, 8H), 1.96 - 1.91 (q, J = 7.6 Hz, J = 13.2 Hz, 2H), 1.64 - 1.55 (m, 8H), 1.40 - 1.28 (m, 48H), 0.92 - 0.87 (m, 12H)

[0337] Project 2: [Chemistry] [2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[(9Z,12Z)-octadeca-9,12-dienoyl]oxypropyl](9Z,2Z)-octadeca-9,12-dienoate (900 mg, 911.38 μmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (598.68 mg, 1. A mixture of 82 mmol, 2 equivalents, EDCI (209.66 mg, 1.09 mmol, 1.2 equivalents), DIPEA (294.47 mg, 2.28 mmol, 396.86 μL, 2.5 equivalents), and DMAP (22.27 mg, 182.28 μmol, 0.2 equivalents) in DCM (9 mL) was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~1 / 1). The compound [2-(4,4-dioctoxybutanoyloxymethyl)-2-[[(9Z,12Z)-octadeca-9,12-dienoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl](9Z,12Z)-octadeca-9,12-dienoate (140 mg, 103.69 μmol, yield 11.38%, purity 96.13%) was obtained as a colorless oil. 1H NMR(400MHz,chloroform-d)δ ppm 5.42-5.30(m,8H),5.18(s,1H),4.75(s,1H),4.49-4.47(t,J=5.2Hz,1H),4.12-4.11(d,J=3.2Hz,8H),3.59-3 .53(m,2H),3.43-3.38(m,2H),3.302-3.289(d,J=5.2Hz,2H),2.79-2.76(t,J=6.8Hz,4H),2.62-2.59(t,J=5.6 Hz,2H),2.53(s,4H),2.42-2.35(m,4H),2.32-2.29(t,J=7.6Hz,4H),2.08-2.03(q,J=6.4Hz,J=13.2Hz,8H),1. 94-1.89(q,J=6.4Hz,J=13.2Hz,3H),1.78(s,3H),1.62-1.53(m,10H),1.40-1.28(m,58H),0.91-0.87(m,15H).

[0338] Example 14 - Compound 7: O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate Step 1: [ka] A mixture of (Z)-nona-3-en-1-ol (50 g, 351.52 mmol, 1 equivalent), heptanediol (281.51 g, 1.76 mol, 5 equivalents), EDCI (80.87 g, 421.83 mmol, 1.2 equivalents), DMAP (4.29 g, 35.15 mmol, 0.1 equivalent), and DIPEA (113.58 g, 878.81 mmol, 153.07 mL, 2.5 equivalents) in 500 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound 7-[(Z)-nona-3-enoxy]-7-oxo-heptanoic acid (140 g, 492.28 mmol, yield 35.01%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.63-10.83(m,1H),5.52-5.49(m,1H),5.35-5.30(m,1H),4.09-4.05(t,J=6.8Hz,2H),2.04-2 .29(m,6H),2.07-2.01(m,2H),1.70-1.62(m,4H),1.43-1.28(m,8H),0.91-0.87(t,J=6.8Hz,3H)

[0339] Step 2: [ka] A mixture of 7-[(Z)-nona-3-enoxy]-7-oxo-heptanoic acid (28.69 g, 100.89 mmol, 1 equivalent), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (8 g, 45.40 mmol, 0.45 equivalents), EDCI (23.21 g, 121.07 mmol, 1.2 equivalents), DIPEA (32.60 g, 252.22 mmol, 43.93 mL, 2.5 equivalents), and DMAP (2.47 g, 20.18 mmol, 0.2 equivalents) in 300 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound O7-[[2,2-dimethyl-5-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]heptanedioate (70 g, 98.74 mmol, yield 65.24%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.49(m,2H),5.35-5.32(m,2H),4.10(s,4H),4.08-4.05(t,J=6.8Hz,4H),3.75(s,4H),2.40-2.28(m,12H),2. 06-2.01(q,J=6.8Hz,J=13.6Hz,4H),1.66-1.62(m,8H),1.42(s,6H),1.37-1.26(m,16H),0.91-0.87(t,J=6.4Hz,6H)

[0340] Step 3: [ka] A solution of O7-[[2,2-dimethyl-5-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]heptanedioate (30 g, 42.32 mmol, 1 equivalent) in THF (300 mL) was slowly added to HCl (3 M, 15.52 mL, 1.1 equivalents), and the mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was added to saturated Na2CO3 (300 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1~1 / 1). Compound O7-[2,2-bis(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (20 g, 29.90 mmol, yield 35.33%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.47(m,2H),5.36-5.30(m,2H),4.14(s,4H),4.08-4.04(t,J=7.2Hz),3.58(s,4H),2.84(s,2H),2.40-2.29 (m,12H),2.06-2.00(q,J=6.4Hz,J=14Hz,4H),1.69-1.60(m,8H),1.38-1.28(m,16H),0.91-0.87(t,J=6.8Hz,6H)

[0341] Step 4: [ka] To a mixture in 150 mL of DCM containing O7-[2,2-bis(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (20 g, 29.90 mmol, 1 equivalent), EDCI (6.88 g, 35.88 mmol, 1.2 equivalents), DIPEA (9.66 g, 74.75 mmol, 13.02 mL, 2.5 equivalents), and DMAP (365.28 mg, 2.99 mmol, 0.1 equivalents), 7-[(Z)-nona-3-enoxy]-7-oxo-heptanoic acid (5.10 g, 17.94 mmol, 0.6 equivalents) in 50 mL of DCM was added. The mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with 600 mL (200 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 10 / 1). Compound O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (8 g, 8.55 mmol, yield 28.61%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.49(m,3H),5.35-5.30(m,3H),4.11(s,6H),4.08-4.05(t,J=7.2Hz,6H),3.52-3.50(d,J=6Hz,2H),2.67-2.64(t,J=13.6Hz ,1H),2.40-2.29(m,18H),2.06-2.01(q,J=6.4Hz,13.6Hz,6H),1.68-1.60(m,12H),1.39-1.27(m,24H),0.91-0.87(t,J=6.8Hz,9H)

[0342] Step 5: [ka] O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (8g, 8.55 mmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid A mixture of (3.37 g, 10.26 mmol, 1.2 equivalents), EDCI (1.97 g, 10.26 mmol, 1.2 equivalents), DIPEA (2.76 g, 21.38 mmol, 3.72 mL, 2.5 equivalents), and DMAP (104.50 mg, 855.37 μmol, 0.1 equivalent) in 80 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 10 / 1). Compound O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (3.4 g, 2.73 mmol, yield 31.91%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.47(m,3H),5.35-5.30(m,3H),5.18(s,1H),4.75(s,1H),4.10(s,8H),4.0 8-4.04(t,J=7.2Hz,6H),3.29-3.28(d,J=5.2Hz,2H),2.61-2.58(t,J=5.6Hz,2H), 2.52(s,4H),2.40-2.28(m,22H),2.06-2.01(q,J=6.4Hz,J=13.6Hz,6H),1.90-1.8 7(m,1H),1.77(s,3H),1.67-1.59(m,14H),1.37-1.27(m,32H),0.91-0.86(m,12H)

[0343] Example 15 - Compound 8: O7-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate Step 1: [ka] To a 15 mL solution of O7-[2,2-bis(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (1.5 g, 2.24 mmol, 1 equivalent) in DCM (15 mL), EDCI (515.87 mg, 2.69 mmol, 1.2 equivalents), DIPEA (724.57 mg, 5.61 mmol, 976.51 μL, 2.5 equivalents) and DMAP (54.79 mg, 448.50 μmol, 0.2 equivalents) were added. Then, a 15 mL solution of 2-(1-adamantyl)acetic acid (392.08 mg, 2.02 mmol, 0.9 equivalents) in DCM (15 mL) was added dropwise to the reaction mixture. The mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 5 / 1). Compound O7-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxoheptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.7 g, 828.26 μmol, yield 36.93%) was obtained as a colorless oil. 1H NMR(400MHz, CDCl3)δ ppm 5.54-5.48(m,2H),5.37-5.31(m,2H),4.12(s,4H),4.09-4.05(t,J=6.4Hz,6H),3.53-3.52(d,J=4Hz,2H),2.61(s,1H),2.40-2.29(m,1 2H),2.10(s,2H),2.07-2.02(q,J=6.8Hz,J=14Hz,4H),1.98(s,3H),1.73-1.60(m,20H),1.40-1.27(m,16H),0.91-0.88(t,J=6.4Hz,6H)

[0344] Project 2:

change

[0345] Example 16 - Compound 9: O7-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate Step 1: [ka] A mixture of O7-[2,2-bis(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (3.2 g, 4.78 mmol, 1 equivalent), 4,4-dioctoxybutanoic acid (824.11 mg, 2.39 mmol, 0.5 equivalent), DMAP (58.45 mg, 478.40 μmol, 0.1 equivalent), DIPEA (1.55 g, 11.96 mmol, 2.08 mL, 2.5 equivalents), and EDCI (1.10 g, 5.74 mmol, 1.2 equivalents) in a DCM (32 mL) was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with 150 mL (50 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~1 / 1). Compound O7-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (1.4 g, 1.41 mmol, yield 29.40%) was obtained as a colorless oil. 11H NMR (400 MHz, CDCl3) δ ppm 5.54 - 5.50 (m, 2H), 5.35 - 5.33 (m, 2H), 4.50 - 4.47 (t, J = 5.2 Hz, 1H), 4.11 (s, 6H), 4.09 - 4.05 (t, J = 7.2 Hz, 4H), 3.59 - 3.51 (m, 4H), 3.43 - 3.38 (m, 2H), 2.67 - 2.64 (t, J = 6.8 Hz, 1H), 2.42 - 2.29 (m, 14H), 2.07 - 2.02 (q, J = 6.8 Hz, J = 14.0 Hz, 4H), 1.96 - 1.91 (q, J = 7.6 Hz, J = 13.2 Hz, 2H), 1.68 - 1.54 (m, 12H), 1.40 - 1.28 (m, 36H), 0.91 - 0.87 (m, 12H)

[0346] Project 2: [Chemistry] O7-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (1.4g, 1.41 mmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (461.95mg) A mixture of 14 mL of DCM containing 1.41 mmol, 1 equivalent, DMAP (17.18 mg, 140.65 μmol, 0.1 equivalent), EDCI (323.55 mg, 1.69 mmol, 1.2 equivalents), and DIPEA (454.44 mg, 3.52 mmol, 612.45 μL, 2.5 equivalents) was degassed, purged three times with N2, and then stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with 150 mL of ethyl acetate (50 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~1 / 1). Compound O7-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (150 mg, 114.87 μmol, yield 8.17%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,2H),5.35-5.32(m,2H),5.19(s,1H),4.74(s,1H),4.49-4.46(t,J=5.6Hz,1H),4.11-4.10(d ,J=2Hz,8H),4.08-4.04(t,J=6.8Hz,4H),3.58-3.53(m,2H),3.43-3.37(m,2H),3.30-3.28(d,J=5.6Hz,2H) ,2.61-2.58(t,J=5.6Hz,2H),2.53(s,4H),2.41-2.28(m,16H),2.06-2.01(q,J=6.8Hz,J=13.6Hz,4H),1.93 -1.88(q,J=7.6Hz,J=13.2Hz,3H),1.78(s,5H),1.67-1.52(m,14H),1.37-1.28(m,44H),0.91-0.86(m,15H)

[0347] Example 17 - Compound 10: O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] A mixture of (Z)-nona-3-en-1-ol (40 g, 281.22 mmol, 1 equivalent), octanedioic acid (244.93 g, 1.41 mol, 5 equivalents), EDCI (70.08 g, 365.58 mmol, 1.3 equivalents), DIPEA (109.04 g, 843.66 mmol, 146.95 mL, 3 equivalents), and DMAP (3.44 g, 28.12 mmol, 0.1 equivalent) in 400 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound 8-[(Z)-nona-3-enoxy]-8-oxo-octanoic acid (140 g, 469.14 mmol, yield 33.36%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.33(s,1H),5.54-5.48(m,1H),5.37-5.31(m,1H),4.09-4.05(t,J=6.8Hz,2H),2.40-2.28(m,6H),2 .07-2.01(q,J=6.4Hz,J=14Hz,2H),1.68-1.60(m,4H),1.39-1.27(m,10H),0.91-0.88(t,J=6.8Hz,3H)

[0348] Step 2: [ka] A mixture of 8-[(Z)-nona-3-enoxy]-8-oxo-octanoic acid (30.11 g, 100.89 mmol, 1 equivalent), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (8 g, 45.40 mmol, 0.45 equivalents), EDCI (23.21 g, 121.07 mmol, 1.2 equivalents), DIPEA (32.60 g, 252.22 mmol, 43.93 mL, 2.5 equivalents), and DMAP (1.23 g, 10.09 mmol, 0.1 equivalent) in 300 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound O8-[[2,2-dimethyl-5-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]octanedioate (70 g, 94.98 mmol, yield 62.76%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.47(m,2H),5.37-5.30(m,2H),4.11(s,4H),4.08-4.05(t,J=6.8Hz,4H),3.74(s,4H),2.40-2.27(m,12H),2. 07-2.01(q,J=7.2Hz,J=14Hz,4H),1.64-1.59(m,8H),1.42(s,6H),1.37-1.27(m,20H),0.91-0.88(t,J=6.8Hz,6H).

[0349] Step 3: [ka] The solution of O8-[[2,2-dimethyl-5-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]octanedioate (70 g, 94.98 mmol, 1 equivalent) and HCl (3 M, 34.83 mL, 1.1 equivalents) in THF (700 mL) was degassed, purged three times with N2, and then stirred at 20°C for 7 hours under an N2 atmosphere. The reaction mixture was slowly added to 500 mL of saturated Na2CO3 at 20°C. This was then extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1~1 / 1). Compound O8-[2,2-bis(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (38 g, 54.52 mmol, yield 57.41%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.53-5.47(m,2H),5.37-5.30(m,2H),4.14(s,1H),4.08-4.05(t,J=7.2Hz,4H),3.58(s,4H),2.81(s,2H),2.40-2.2 8(m,12H),2.06-2.01(q,J=6.8Hz,J=14Hz,4H),1.65-1.60(m,8H),1.37-1.27(m,20H),0.90-0.87(t,J=6.4Hz,6H).

[0350] Step 4: [ka] To a solution of O8-[2,2-bis(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (23 g, 33.00 mmol, 1 equivalent), EDCI (7.59 g, 39.60 mmol, 1.2 equivalents), DIPEA (10.66 g, 82.50 mmol, 14.37 mL, 2.5 equivalents), and DMAP (403.17 mg, 3.30 mmol, 0.1 equivalents) in DCM (230 mL), 8-[(Z)-nona-3-enoxy]-8-oxo-octanoic acid (5.91 g, 19.80 mmol, 0.6 equivalents) in DCM (230 mL) was added. The mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with 800 mL (400 mL x 2) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 10 / 1). Compound O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (11 g, 11.25 mmol, yield 34.10%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.48(m,3H),5.37-5.31(m,3H),4.11(s,6H),4.09-4.05(t,J=6.8Hz,6H),3.51-3.50(d,J=6.8Hz,2H),2.61-2.57(t,J=6.8Hz,1H) ,2.40-2.28(m,18H),2.07-2.02(q,J=6.8Hz,J=14Hz,6H),1.64-1.61(t,J=7.2Hz,12H),1.38-1.27(m,30H),0.91-0.88(t,J=6.4Hz,9H)

[0351] Step 5: [ka] O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (11g, 11.25mmol, 1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)deca A mixture of nic acid (3.33 g, 10.13 mmol, 0.9 equivalents), EDCI (2.59 g, 13.51 mmol, 1.2 equivalents), DIPEA (3.64 g, 28.14 mmol, 4.90 mL, 2.5 equivalents), and DMAP (137.50 mg, 1.13 mmol, 0.1 equivalent) in 100 mL of DCM was degassed, purged three times with N2, and then stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with water (200 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). Compound O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (3.5 g, 2.72 mmol, yield 24.15%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.54-5.48(m,3H),5.37-5.31(m,3H),5.17(s,1H),4.75(s,1H),4.11(s,8H),4.08-4.05(t,J=6 .8Hz,8H),3.29-3.28(d,J=4.8Hz,2H),2.60-2.57(t,J=6.0Hz,4H),2.52(s,4H),2.40-2.35(q, J=7.2Hz,J=14.0Hz,8H),2.33-2.28(q,J=7.2Hz,J=12.4Hz,12H),2.07-2.01(q,J=6.8Hz,J=14. 0Hz,6H),1.89(s,1H),1.78(s,3H),1.67-1.59(m,14H),1.39-1.28(m,40H),0.91-0.86(m,12H).

[0352] Example 18 - Compound 11: O8-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] To a 10 mL solution of O8-[2,2-bis(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (1 g, 1.43 mmol, 1 equivalent) in DCM (10 mL), DIPEA (463.60 mg, 3.59 mmol, 624.80 μL, 2.5 equivalents), EDCI (330.07 mg, 1.72 mmol, 1.2 equivalents), DMAP (17.53 mg, 143.48 μmol, 0.1 equivalent), and 2-(1-adamantyl)acetic acid (278.74 mg, 1.43 mmol, 1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was poured into H2O (50 mL) and extracted with DCM (50 mL x 3). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1 to 5 / 1). Compound O8-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (450 mg) was obtained as a colorless oil.

[0353] Step 2: [ka] O8-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanediate (0.65g, 744.39μmol, 1 equivalent) in a DCM (6.5mL) solution with EDCI (171 (0.24 mg, 893.26 μmol, 1.2 equivalents), DMAP (9.09 mg, 74.44 μmol, 0.1 equivalent), DIPEA (240.52 mg, 1.86 mmol, 324.15 μL, 2.5 equivalents), and 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (293.39 mg, 893.26 μmol, 1.2 equivalents) were added. This mixture was stirred under N2 at 15°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1~2 / 1). Compound O8-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (200 mg) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.51-5.47(m,2H),5.37-5.30(m,2H),5.17(s,1H),4.74(m,1H),4.11-4.04(m,12H),3.2 9-3.28(d,J=4.0Hz,2H),2.60-2.57(t,J=4.0Hz,2H),2.52(s,4H),2.39-2.34(q,J=4.0Hz ,J=12.0Hz,6H),2.32-2.27(m,8H),2.07-2.00(m,6H),1.96(s,3H),1.88-1.87(m,1H),1. 78(s,5H),1.77-1.68(m,9H),1.62-1.57(m,16H),1.35-1.26(m,30H),0.90-0.86(m,9H).

[0354] Example 19 - Compound 12: O8-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] To a 15 mL solution of O8-[2,2-bis(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (1.5 g, 2.15 mmol, 1 equivalent) in DCM (15 mL), DIPEA (695.40 mg, 5.38 mmol, 937.20 μL, 2.5 equivalents), EDCI (495.11 mg, 2.58 mmol, 1.2 equivalents), DMAP (26.29 mg, 215.22 μmol, 0.1 equivalent), and 4,4-dioctoxybutanoic acid (444.90 mg, 1.29 mmol, 0.6 equivalents) were added. This mixture was stirred under N2 conditions at 20 °C for 12 hours. The reaction mixture was poured into H2O (50 mL) and extracted with DCM (50 mL x 3). The combined organic layer was washed with brine (50 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1~5 / 1). Compound O8-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (0.7 g) was obtained as a colorless oil.

[0355] Step 2: [ka] O8-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]octanedioate (700 mg, 683.95 μmol, 1 equivalent) in a DCM (7 mL) solution with DIPEA (220. 99 mg, 1.71 mmol, 297.83 μL, 2.5 equivalents) of EDCI (157.34 mg, 820.74 μmol, 1.2 equivalents), DMAP (8.36 mg, 68.40 μmol, 0.1 equivalent), and 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (269.57 mg, 820.74 μmol, 1.2 equivalents) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was poured into H2O (50 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1 to 5 / 1). Compound O8-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (160 mg) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ=5.54-5.48(m,2H),5.38-5.31(m,2H),5.17(s,1H),4.75(s,1H),4.50-4.47(t,J=8.0Hz ,1H),4.12-4.11(d,J=4.0Hz,8H),4.09-4.05(t,J=6.8Hz,4H),3.59-3.53(m,2H),3.43-3.38(m,2H),3.30-3. 28(m,2H),2.61-2.58(t,J=6.0Hz,2H),2.52(s,4H),2.42-2.35(m,8H),2.33-2.28(m,8H),2.07-2.02(q,J=6. 8Hz,J=14Hz,4H),1.94-1.89(m,3H),1.78(s,5H),1.64-1.55(m,14H),1.38-1.28(m,48H),0.91-0.87(m,15H)

[0356] Example 20 - Compound 13: O6-[2,2-bis[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]hexanedioate Step 1: [ka] To a solution of (Z)-nona-3-en-1-ol (30.00 g, 210.91 mmol, 1 equivalent) in DCM (300 mL), EDCI (48.52 g, 253.10 mmol, 1.2 equivalents), DIPEA (68.15 g, 527.29 mmol, 91.84 mL, 2.5 equivalents), adipic acid (154.12 g, 1.05 mol, 175.13 mL, 5 equivalents) and DMAP (2.58 g, 21.09 mmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (300 mL) and extracted with 900 mL (300 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 6-[(Z)-nona-3-enoxy]-6-oxohexanoic acid (30 g, 110.96 mmol, yield 52.61%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.50(m,1H),5.35-5.32(m,1H),4.09-4.06(t,J=7.2Hz,2H),2.04-2.32(m,6H) ,2.06-2.01(m,2H),1.70-1.67(m,4H),1.37-1.26(m,6H),0.91-0.88(t,J=6.8Hz,3H)

[0357] Step 2: [ka] To a solution of 6-[(Z)-nona-3-enoxy]-6-oxohexanoic acid (26.85 g, 99.31 mmol, 1 equivalent) in DCM (270 mL), EDCI (22.85 g, 119.18 mmol, 1.2 equivalents), DIPEA (32.09 g, 248.28 mmol, 43.25 mL, 2.5 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (7 g, 39.73 mmol, 0.4 equivalents), and DMAP (1.21 g, 9.93 mmol, 0.1 equivalents) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (300 mL) and extracted with 900 mL (300 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[[2,2-dimethyl-5-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]hexanedioate (15 g, 22.03 mmol, yield 22.18%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,2H),5.35-5.30(m,2H),4.11(S,1H),4.09-4.05(t,J=7.2Hz,4H),3.74(s,4H),2.40-2.30(m,12H),2. 06-2.01(q,J=6.8Hz,J=14Hz,4H),1.67-1.64(m,8H),1.42(s,6H),1.37-1.26(m,12H),0.91-0.88(t,J=6.4Hz,6H).

[0358] Step 3: [ka] O6-[[2,2-dimethyl-5-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]hexanedioate (15 g, 22.03 mmol, 1 equivalent) was dissolved in THF (150 mL) and HCl (3 M, 8.08 mL, 1.1 equivalents) was slowly added. This mixture was stirred under N2 at 20 °C for 7 hours. The reaction mixture was diluted with saturated NaHCO3 (300 mL) and extracted with DCM 900 mL (300 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). The compound O6-[2,2-bis(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (6 g, 9.36 mmol, yield 42.50%) was obtained as a colorless oil.

[0359] Step 4: [ka] To a solution of O6-[2,2-bis(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxohexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (2 g, 3.12 mmol, 1 equivalent) in DCM (20 mL), EDCI (717.93 mg, 3.75 mmol, 1.2 equivalents), DIPEA (1.01 g, 7.80 mmol, 1.36 mL, 2.5 equivalents), 6-[(Z)-nona-3-enoxy]-6-oxohexanoic acid (506.27 mg, 1.87 mmol, 0.6 equivalents) and DMAP (38.13 mg, 312.09 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM 90 mL (30 mL x 3). The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[2-(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxy-2-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.9 g, 1.01 mmol, yield 32.29%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.48(m,3H),5.37-5.32(m,3H),4.12(s,6H),4.09-4.05(t,J=7.2Hz,6H),3.53-3.51(d,J=6.4Hz,2H),2.69-2.67(t ,J=6.4Hz,1H),2.40-2.31(m,18H),2.07-2.01(m,6H),1.68-1.64(m,12H),1.38-1.27(m,18H),0.91-0.88(t,J=6.8Hz,9H)

[0360] Step 5: [ka] O6-[2-(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxy-2-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.9g, 1.01 mmol, 1 equivalent) in DCM (9mL) solution with EDCI (231 (0.79 mg, 1.21 mmol, 1.2 equivalents), DIPEA (325.57 mg, 2.52 mmol, 438.77 μL, 2.5 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (397.14 mg, 1.21 mmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (14.93 mg, 100.76 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM 90 mL (30 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[2,2-bis[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.5 g, 415.41 μmol, yield 41.23%) was obtained as a colorless oil. 1 ¹H NMR (400 MHz, CDCl3) δ ppm 5.52-5.48(m,3H),5.37-5.19(m,3H),5.19(s,1H),4.75(s,1H),4.11(s,8 H),4.09-4.05(t,J=6.8Hz,6H),3.23-3.29(d,J=4Hz,2H),2.60(s,2H),2.5 2(s,4H),2.40-2.31(m,20H),2.07-2.01(q,J=6.8Hz,6H),1.92-1.85(m,1H ),1.78(s,5H),1.67-1.63(m,14H),1.38-1.27(m,26H),0.91-0.86(m,12H)

[0361] Example 21 - Compound 14: O6-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]hexanedioate Step 1: [ka] To a 20 mL solution of O6-[2,2-bis(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (2 g, 3.12 mmol, 1 equivalent) in DCM (20 mL), EDCI (717.94 mg, 3.75 mmol, 1.2 equivalents), DIPEA (1.01 g, 7.80 mmol, 1.36 mL, 2.5 equivalents), 2-(1-adamantyl)acetic acid (363.78 mg, 1.87 mmol, 0.6 equivalents) and DMAP (38.13 mg, 312.09 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 conditions at 20 °C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[2-[[2-(1-adamantyl)acetyl]oxymethyl]-2-(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.8 g, 979.08 μmol, yield 31.37%) was obtained as a colorless oil. 1H NMR(400MHz, CDCl3)δ ppm 5.52-5.48(m,2H),5.37-5.31(m,2H),4.12(s,4H),4.09-4.06(m,6H),5.53-5.52(d,J=6.8Hz,2H),2.63-2.59(t,J=6.8Hz,1H),2.40-2.31( m,12H),2.10(s,2H),2.07-2.01(q,J=6.4Hz,J=14Hz,4H),1.98(s,3H),1.73-1.59(m,20H),1.38-1.27(m,12H),0.91-0.88(t,J=6.4Hz,6H)

[0362] Project 2:

change

[0363] Example 22 - Compound 15: O6-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]hexanedioate Step 1: [ka] To a 20 mL solution of O6-[2,2-bis(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (2 g, 3.12 mmol, 1 equivalent) in DCM (20 mL), EDCI (717.93 mg, 3.75 mmol, 1.2 equivalents), 4,4-dioctoxybutanoic acid (645.14 mg, 1.87 mmol, 0.6 equivalents), DIPEA (1.01 g, 7.80 mmol, 1.36 mL, 2.5 equivalents) and DMAP (38.13 mg, 312.09 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20 °C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.6 g, 620.25 μmol, yield 19.87%) was obtained as a colorless oil.

[0364] Step 2: [ka] O6-[2-(4,4-dioctoxybutanoyloxymethyl)-2-(hydroxymethyl)-3-[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.6g, 620.25μmol, 1 equivalent) in DCM (6mL) solution with EDCI (142.68mg, 74 4.30 μmol, 1.2 equivalents) of 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (305.58 mg, 930.37 μmol, 1.5 equivalents), DIPEA (200.41 mg, 1.55 mmol, 270.09 μL, 2.5 equivalents), and 4-pyrrolidine-1-ylpyridine (9.19 mg, 62.02 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (10 mL) and extracted with 30 mL (10 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O6-[2-(4,4-dioctoxybutanoyloxymethyl)-2-[[6-[(Z)-nona-3-enoxy]-6-oxo-hexanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]hexanedioate (0.15 g, 117.39 μmol, yield 18.93%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.54-5.48(m,2H),5.35-5.33(m,2H),5.19(s,1H),4.75(s,1H),4.50-4.47(t,J=5.6Hz,1H),4.11(s,8H),4 .09-4.05(t,J=7.2Hz,4H),3.57-3.53(m,2H),3.43-3.38(m,2H),3.30-3.28(d,J=5.2Hz,2H),2.61-2.58(t, J=5.6Hz,2H),2.52(s,4H),2.40-2.32(m,14H),2.07-2.02(q,J=6.4Hz,J=14Hz,4H),1.94-1.89(q,J=7.6Hz ,J=13.6Hz,3H),1.78(s,5H),1.69-1.63(m,10H),1.60-1.53(m,6H),1.36-1.28(m,40H),0.91-0.87(m,15H)

[0365] Example 23 - Compound 16: O7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate Step 1: [ka] To a solution of heptane-1,7-diol (49.50 g, 374.41 mmol, 3 equivalents) in DCM (500 mL), DIPEA (40.32 g, 312.01 mmol, 54.35 mL, 2.5 equivalents), 2-butyloctanoic acid (25 g, 124.80 mmol, 1 equivalent), DMAP (1.52 g, 12.48 mmol, 0.1 equivalent), and EDCI (28.71 g, 149.76 mmol, 1.2 equivalents) were added. This mixture was stirred under N2 at 25°C for 12 hours. The reaction mixture was diluted with H2O (600 mL) and extracted with 1000 mL (500 mL x 2) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound 7-hydroxyheptyl 2-butyloctanoate (100 g, 317.96 mmol, yield 63.69%) was obtained as a colorless oil.

[0366] Step 2: [ka] CrO3\H2SO4 (238.47 mg, 119.24 mmol, 1.5 equivalents) was added dropwise at 0°C to a solution of 7-hydroxyheptyl-2-butyloctanoate (25 g, 79.49 mmol, 1 equivalent) in acetone (250 mL). The mixture was stirred under N2 at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 300 mL (100 mL x 3) of siRNA. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound 7-(2-butyloctanoyloxy)heptanoic acid (50 g, 152.21 mmol, yield 47.87%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.27-10.80(m,1H),4.09-4.06(t,J=6.8Hz,2H),2.39-2.28(m,3H),1.5 7-1.55(m,6H),1.47-1.37(m,6H),1.33-1.22(m,12H),0.90-0.86(m,6H)

[0367] Step 3: [ka] To a 20 mL solution of O7-[2,2-bis(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (2 g, 2.99 mmol, 1 equivalent) in DCM (20 mL), EDCI (687.83 mg, 3.59 mmol, 1.2 equivalents), DIPEA (966.09 mg, 7.48 mmol, 1.30 mL, 2.5 equivalents), 7-(2-butyloctanoyloxy)heptanoic acid (589.30 mg, 1.79 mmol, 0.6 equivalents) and DMAP (36.53 mg, 299.00 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 conditions at 20 °C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 30 mL (10 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (1 g, 1.02 mmol, yield 34.15%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.48(m,2H),5.37-5.33(m,2H),4.12(s,6H),4.09-4.05(t,J=6.8Hz,6H),3.52-3.50(d,J=6.8Hz,2H),2.64-2.61(t,J=6.8Hz,1H), 2.39-2.29(m,14H),2.07-2.02(q,J=6.4Hz,J=10.8Hz,4H),1.68-1.57(m,16H),1.47-1.43(m,2H),1.38-1.26(m,30H),0.91-0.86(m,12H)

[0368] Step 4: [ka] O7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (1g, 1.02 mmol, 1 equivalent) in DCM (10 mL) solution with EDCI (23 4.89 mg (1.23 mmol, 1.2 equivalents), DIPEA (329.92 mg, 2.55 mmol, 444.63 μL, 2.5 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (301.83 mg, 918.96 μmol, 0.9 equivalents), and DMAP (12.47 mg, 102.11 μmol, 0.1 equivalents) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 30 mL (10 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.15 g, 116.30 μmol, yield 11.39%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.48(m,2H),5.37-5.31(m,2H),5.27-5.01(m,1H),4.75(s,1H) ,4.11(s,8H),4.08-4.05(t,J=6.8Hz,6H),3.31(s,2H),2.62-2.55(m, 6H),2.40-2.29(m,18H),2.07-2.02(q,J=6.4Hz,J=14Hz,4H),1.91-1. 80(m,5H),1.66-1.59(m,14H),1.46-1.27(m,44H),0.91-0.86(m,15H)

[0369] Example 24 - Compound 17: O7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate Step 1: [ka] To a solution of 7-(2-butyloctanoyloxy)heptanoic acid (24.86 g, 75.67 mmol, 1 equivalent) in DCM (250 mL), EDCI (17.41 g, 90.80 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (6 g, 34.05 mmol, 0.45 equivalents), DMAP (924.41 mg, 7.57 mmol, 0.1 equivalent), and DIPEA (24.45 g, 189.17 mmol, 32.95 mL, 2.5 equivalents) were added. This mixture was stirred under N2 at 25°C for 12 hours. The mixture was diluted with H2O (300 mL) and extracted with 500 mL (100 mL x 5) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound [7-[[5-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methoxy]-7-oxo-heptyl]2-butyloctanoate (13 g, 16.31 mmol, yield 21.55%) was obtained as a colorless oil.

[0370] Step 2: [ka] [7-[[5-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methoxy]-7-oxo-heptyl]2-butyloctanoate (27 g, 33.87 mmol, 1 equivalent) was dissolved in THF (270 mL) and HCl (3 M, 12.42 mL, 1.1 equivalents) was added dropwise at 0°C. The mixture was stirred under N2 at 25°C for 5 hours. This was then poured into H2O (200 mL) and extracted with DCM 800 mL (200 mL x 4). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The compound [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxy-2-(hydroxymethyl)propoxy]-7-oxo-heptyl]2-butyloctanoate (10 g, 13.21 mmol, yield 39.00%) was obtained as a colorless oil.

[0371] Step 3: [ka] To a 20 mL solution of [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxy-2-(hydroxymethyl)propoxy]-7-oxo-heptyl]2-butyloctanoate (2 g, 2.64 mmol, 1 equivalent) in DCM (20 mL), EDCI (1.01 g, 5.28 mmol, 2 equivalents), 7-[(Z)-nona-3-enoxy]-7-oxo-heptanoic acid (450.77 mg, 1.59 mmol, 0.6 equivalents), DMAP (32.27 mg, 264.17 μmol, 0.1 equivalent), and DIPEA (682.84 mg, 5.28 mmol, 920.28 μL, 2 equivalents) were added. This mixture was stirred under N2 conditions at 25 °C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM 150 mL (50 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound O7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.7 g, 683.95 μmol, yield 25.89%) was obtained as a colorless oil.

[0372] Step 4: [ka] To a solution of O7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxypropyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.7 g, 683.95 μmol, 1.2 equivalents) in DCM (7 mL), EDCI (218.53 mg, 1.14 mmol, 2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (187.20 mg, 569.96 μmol, 1 equivalent), DMAP (6.96 mg, 57.00 μmol, 0.1 equivalent), and DIPEA (147.33 mg, 1.14 mmol, 198.55 μL, 2 equivalents) were added. This mixture was stirred under N2 conditions at 25°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound O7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (111 mg) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.48(m,1H),5.43-5.04(m,2H),4.75(s,1H),4.11(s,8H),4.08-4.05(t,J=6.8Hz,6H),3.34(s,2H),2.67-2.60(m,6H),2.40- 2.28(m,14H),2.07-2.01(q,J=6.8Hz,J=13.6Hz,2H),1.90-1.76(m,6H),1.63-1.56(m,16H),1.47-1.24(m,54H),0.91-0.86(m,18H)

[0373] Example 25 - Compound 18: [7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate Step 1: [ka] [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxyl-2-(hydroxymethyl)propoxy]-7-oxo-heptyl]2-butyloctanoate (3 g, 3.96 mmol, 1 equivalent) was dissolved in DCM (30 mL) and EDCI (911.56 mg, 4.76 mmol, 1.2 equivalents), DIPEA (51.21 mg, 396.26 μmol, 69.02 μL, 0.1 equivalent), 7-(2-butyloctanoyloxy)heptanoic acid (780.99 mg, 2.38 mmol, 0.6 equivalents), and DMAP (1.21 g, 9.91 mmol, 2.5 equivalents) were added. The mixture was stirred under N2 at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM 150 mL (50 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound [7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxy-propoxy]-7-oxo-heptyl]2-butyloctanoate (1.5 g, 1.41 mmol, yield 35.46%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.12(s,6H),4.09-4.06(t,J=6.8Hz,6H),3.515-3.50(d,J=6Hz),2.59-2.56(t,J=6.4 Hz,1H),2.36-2.28(m,9H),1.67-1.55(m,24H),1.45-1.26(m,48H),0.91-0.87(m,18H)

[0374] Step 2: [ka] To a 13 mL solution of [7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxy-propoxy]-7-oxo-heptyl]2-butyloctanoate (1.3 g, 1.22 mmol, 1 equivalent) in DCM (13 mL), EDCI (466.88 mg, 2.44 mmol, 2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (359.96 mg, 1.10 mmol, 0.9 equivalents), DMAP (14.88 mg, 121.77 μmol, 0.1 equivalent), and DIPEA (314.77 mg, 2.44 mmol, 424.21 μL, 2 equivalents) were added. This mixture was stirred under N2 at 25 °C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 100 mL (50 mL x 2) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound [7-[2,2-bis[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate (106 mg, 76.92 μmol, yield 6.32%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 0.86-0.90(td,J=6.94,2.2Hz,21H)1.23-1.33(m,42H)1.34-1.51(m,20H)1.51-1.69(m,20H)1.78(m,6H)1.84-1.9 6(m,1H)2.30-2.42(m,10H)2.53(m,4H)2.60(m,2H)3.29(d,J=4.8Hz,2H)4.07(t,J=6.6Hz,6H)4.12(s,8H)4.75(br s,1H)5.18(br s,1H).

[0375] Example 26 - Compound 19: [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(4,4-dioctoxybutanoyloxymethyl)-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate Step 1: [ka] To a 20 mL solution of DCM containing [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-3-hydroxy-2-(hydroxymethyl)propoxy]-7-oxoheptyl]2-butyloctanoate (2 g, 2.64 mmol, 1 equivalent), EDCI (607.70 mg, 3.17 mmol, 1.2 equivalents), DIPEA (853.56 mg, 6.60 mmol, 1.15 mL, 2.5 equivalents), and DMAP (32.27 mg, 264.17 μmol, 0.1 equivalent), 4,4-dioctoxybutanoic acid (728.11 mg, 2.11 mmol, 0.8 equivalents) in a 20 mL solution of DCM was added dropwise to the reaction mixture. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 5 / 1). Compound [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(4,4-dioctoxybutanoyloxymethyl)-3-hydroxypropoxy]-7-xo(xo)-heptyl]2-butyloctanoate (1.4 g, 1.29 mmol, yield 48.91%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,chloroform-d)δ ppm 0.70-0.90(m,18H)1.12-1.43(m,58H)1.48-1.62(m,14H)1.79-1.92(m,2H)2.16-2.29(m,6H)2.34(t,J=7.6Hz,2H)2.5 4(t,J=7.0Hz,1H)3.33(dt,J=9.2,6.8Hz,2H)3.40-3.57(m,4H)3.99(t,J=6.6Hz,4H)4.04(s,6H)4.41(t,J=5.4Hz,1H).

[0376] Step 2: [ka] To a 10 mL solution of DCM (Digital Cholesterol) containing [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(4,4-dioctoxybutanoyloxymethyl)-3-hydroxypropoxy]-7-oxo-heptyl]2-butyloctanoate (1 g, 922.85 μmol, 1 equivalent), EDCI (212.29 mg, 1.11 mmol, 1.2 equivalents), DIPEA (298.18 mg, 2.31 mmol, 401.86 μL, 2.5 equivalents), and DMAP (11.27 mg, 92.29 μmol, 0.1 equivalent), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (272.80 mg, 830.57 μmol, 0.9 equivalents) was added to the above reaction mixture. The mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 30 mL (10 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1~1 / 1). The compound [7-[2-[7-(2-butyloctanoyloxy)heptanoyloxymethyl]-2-(4,4-dioctoxybutanoyloxymethyl)-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate (0.102 g, 73.17 μmol, yield 7.93%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 0.86-0.90(td,J=7.0,2.4Hz,6H)1.22-1.32(m,12H)1.36-1.45(m,12H)1.59-1.67(m,6H )2.35-2.38(m,4H)3.50(s,2H)3.70-3.76(m,4H)4.05-4.09(t,J=6.6Hz,2H)4.25(s,2H).

[0377] Example 27 - Compound 20: [7-[2,2-bis(4,4-dioctoxybutanoyloxymethyl)-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate Step 1: [ka] To a solution of 7-(2-butyloctanoyloxy)heptanoic acid (5.83 g, 17.73 mmol, 1 equivalent) in DCM (58 mL), EDCI (4.08 g, 21.28 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (5 g, 28.38 mmol, 1.6 equivalents), DIPEA (5.73 g, 44.34 mmol, 7.72 mL, 2.5 equivalents), and DMAP (216.66 mg, 1.77 mmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). The compound [7-[[5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methoxy]-7-oxo-heptyl]2-butyloctanoate (3g, 6.16 mmol, yield 34.76%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 0.86-0.90(td,J=7.0,2.4Hz,6H)1.22-1.32(m,12H)1.36-1.45(m,12H)1.59-1.67(m,6H )2.35-2.38(m,4H)3.50(s,2H)3.70-3.76(m,4H)4.05-4.09(t,J=6.6Hz,2H)4.25(s,2H).

[0378] Step 2: [ka] [7-[[5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methoxy]-7-oxoheptyl]2-butyloctanoate (3g, 6.16 mmol, 1 equivalent) was dissolved in THF (30 mL) and HCl (3M, 2.26 mL, 1.1 equivalents) was slowly added at 0°C. This mixture was stirred under N2 at 20°C for 7 hours. The reaction mixture was added to saturated NaHCO3 to adjust the pH to 9 and extracted with 90 mL (30 mL x 3) of ethyl acetate. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). The compound [7-[3-hydroxy-2,2-bis(hydroxymethyl)propoxy]-7-oxo-heptyl]2-butyloctanoate (1.5 g, 3.36 mmol, yield 54.49%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 0.86-0.90(td,J=6.94,2.6Hz,6H)1.21-1.47(m,19H)1.63-1.66(m,6H)2.32(s,1H)2.36-2 .39(t,J=7.4Hz,2H)2.53-2.65(m,2H)3.66(s,6H)4.06-4.09(t,J=6.6Hz,2H)4.23(s,2H).

[0379] Step 3: [ka] To a solution of [7-[3-hydroxy-2,2-bis(hydroxymethyl)propoxy]-7-oxo-heptyl]2-butyloctanoate (1.5 g, 3.36 mmol, 1 equivalent) in DCM (30 mL), EDCI (772.62 mg, 4.03 mmol, 1.2 equivalents), 4,4-dioctoxybutanoic acid (2.31 g, 6.72 mmol, 2 equivalents), DIPEA (1.09 g, 8.40 mmol, 1.46 mL, 2.5 equivalents) and DMAP (41.03 mg, 335.86 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). The compound [7-[2,2-bis(4,4-dioctoxybutanoyloxymethyl)-3-hydroxy-propoxy]-7-oxo-heptyl]2-butyloctanoate (1.2 g, 1.09 mmol, yield 32.49%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 0.87-0.91(m,18H)1.28-1.38(m,56H)1.53-1.65(m,16H)1.91-1.96(m,4H)2.29-2.35(m,3H)2.40-2.44(t,J=7.4Hz,4H)2.64-2.67(t ,J=7.0Hz,1H)3.38-3.44(dt,J=9.2,6.8Hz,4H)3.51-3.59(m,6H)4.05-4.09(t,J=6.6Hz,2H)4.11(s,6H)4.47-4.50(t,J=5.4Hz,2H).

[0380] Step 4: [ka] To a solution of [7-[2,2-bis(4,4-dioctoxybutanoyloxymethyl)-3-hydroxy-propoxy]-7-oxo-heptyl]2-butyloctanoate (1.2 g, 1.09 mmol, 1 equivalent) in DCM (24 mL), EDCI (251.04 mg, 1.31 mmol, 1.2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (322.58 mg, 982.14 μmol, 0.9 equivalents), DIPEA (141.04 mg, 1.09 mmol, 190.08 μL, 2.5 equivalents) and DMAP (13.33 mg, 109.13 μmol, 0.1 equivalent) were added. This mixture was stirred under N2 at 20°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound [7-[2,2-bis(4,4-dioctoxybutanoyloxymethyl)-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propoxy]-7-oxo-heptyl]2-butyloctanoate (0.12 g, 85.10 μmol, yield 7.80%) was obtained as a colorless oil. 1 ¹H NMR (400 MHz, CDCl3) δ ppm 0.81-0.94(m,21H)1.24-1.51(m,66H)1.52-1.66(m,18H)1.79(m,4H)1 .89-1.95(m,4H)2.25-2.43(m,9H)2.53(m,4H)2.61(m,2H)3.20-3.34( m,2H)3.41(dt,J=9.2,6.8Hz,4H)3.56(dt,J=9.2,6.8Hz,4H)4.07(t,J=6.6Hz,2H)4.12(s,8H)4.48(t,J=5.4Hz,2H)4.68-4.90(m,1H)5.20(br s,1H).

[0381] Example 28 - Compound 21: O9-[2,2-bis[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-octa-3-enyl]nonanedioate Step 1: [ka] To a solution of (Z)-octa-3-en-1-ol (20 g, 155.99 mmol, 1 equivalent) in DCM (100 mL) and THF (100 mL), EDCI (35.88 g, 187.19 mmol, 1.2 equivalents), nonanediol (146.80 g, 779.96 mmol, 5 equivalents), DIPEA (50.40 g, 389.98 mmol, 67.93 mL, 2.5 equivalents), and DMAP (1.91 g, 15.60 mmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 9-[(Z)-octa-3-enoxy]-9-oxo-nonanoic acid (20 g, 67.02 mmol, yield 42.96%) was obtained as a colorless oil.

[0382] Step 2: [ka] To a solution of 9-[(Z)-octa-3-enoxy]-9-oxononanoic acid (11.29 g, 37.83 mmol, 1 equivalent) in DCM (110 mL), EDCI (8.70 g, 45.40 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (3 g, 17.03 mmol, 0.45 equivalents), DIPEA (12.22 g, 94.58 mmol, 16.47 mL, 2.5 equivalents), and DMAP (462.21 mg, 3.78 mmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O9-[[2,2-dimethyl-5-[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-octa-3-enyl]nonanedioate (7 g, 9.50 mmol, yield 25.10%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,2H),5.35-5.32(m,2H),4.10(s,4H),4.06(t,J=12.0Hz,8H),3.74(s,4H),2.40-2.35(m,4H),2. 33-2.27(m,4H),2.05-2.02(m,4H),1.60(t,J=8.0Hz,9H),1.42(s,6H),1.34-1.31(m,20H),0.92-0.88(m,6H).

[0383] Step 3: [ka] O9-[[2,2-dimethyl-5-[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-octa-3-enyl]nonanedioate (7 g, 9.50 mmol, 1 equivalent) was dissolved in THF (70 mL) and HCl (3 M, 3.48 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 20°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with NaHCO3 aqueous solution (70 mL) and extracted with DCM 300 mL (100 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). Compound O9-[2,2-bis(hydroxymethyl)-3-[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxypropyl]O1-[(Z)-octa-3-enyl]nonanedioate (2.5 g, 3.59 mmol, yield 37.77%) was obtained as a colorless oil.

[0384] Step 4: [ka] To a 10 mL solution of O9-[2,2-bis(hydroxymethyl)-3-[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxypropyl]O1-[(Z)-octa-3-enyl]nonanediate (1 g, 1.43 mmol, 1 equivalent) in DCM (10 mL), EDCI (330.07 mg, 1.72 mmol, 1.2 equivalents), 9-[(Z)-octa-3-enoxy]-9-oxo-nonanoic acid (256.91 mg, 860.90 μmol, 0.6 equivalents), DIPEA (463.60 mg, 3.59 mmol, 624.80 μL, 2.5 equivalents) and DMAP (17.53 mg, 143.48 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (10 mL) and extracted with 30 mL (10 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O9-[2-(hydroxymethyl)-3-[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxy-2-[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]propyl]O1-[(Z)-octa-3-enyl]nonanedioate (0.5 g, 511.59 μmol, yield 35.66%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,3H),5.36-5.33(m,3H),4.11(s,6H),4.06(t,J=4.0Hz,6H),3.51(d,J=4.0Hz,2H),2. 60(t,J=4.0Hz,1H),2.40-2.27(m,18H),2.07-2.02(m,6H),1.34-1.32(m,28H),0.92-0.89(m,9H).

[0385] Step 5: [ka] O9-[2-(hydroxymethyl)-3-[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxy-2-[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]propyl]O1-[(Z)-octa-3-enyl]nonanedioate (0.5g, 511.59μmol, 1 equivalent) in a DCM (5mL) solution with EDCI (117.69m g (613.91 μmol, 1.2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (201.63 mg, 613.91 μmol, 1.2 equivalents), DIPEA (165.30 mg, 1.28 mmol, 222.77 μL, 2.5 equivalents), and 4-pyrrolidine-1-ylpyridine (7.58 mg, 51.16 μmol, 0.1 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (10 mL) and extracted with DCM 30 mL (10 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. Compound O9-[2,2-bis[[9-[(Z)-octa-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-octa-3-enyl]nonanedioate (133 mg) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,3H),5.36-5.33(m,3H),5.26-5.25(m,1H),4.75(s,1H),4. 12(d,J=3.2Hz,8H),4.06(t,J=7.2Hz,6H),3.32(d,J=4.4Hz,2H),2.61(d, J=25.6Hz,6H),2.40-2.35(m,8H),2.32-2.27(m,12H),2.08-2.03(m,6H), 1.81(s,6H),1.63-1.49(m,16H),1.36-1.27(m,36H),0.92-0.86(m,12H).

[0386] Example 29 - Compound 22: O9-[2,2-bis[[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]nonanedioate Step 1: [ka] To a solution of (Z)-nona-3-en-1-ol (20 g, 140.61 mmol, 1 equivalent) in DCM (100 mL) and THF (100 mL), EDCI (32.35 g, 168.73 mmol, 1.2 equivalents), nonanedioic acid (132.33 g, 703.05 mmol, 5 equivalents), DIPEA (45.43 g, 351.52 mmol, 61.23 mL, 2.5 equivalents), and DMAP (1.72 g, 14.06 mmol, 0.1 equivalent) were added under an N2 atmosphere. This mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (300 mL) and extracted with 900 mL (300 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 9-[(Z)-nona-3-enoxy]-9-oxo-nonanoic acid (20 g, 64.01 mmol, yield 45.52%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.21(br s,1H),5.56-5.45(m,1H),5.39-5.29(m,1H),4.07(t,J=6.8Hz,2H),2.42-2.27(m,6 H),2.08-1.99(m,2H),1.71-1.56(m,4H),1.38-1.20(m,12H),0.89(t,J=6.8Hz,3H).

[0387] Step 2: [ka] To a solution of 9-[(Z)-nona-3-enoxy]-9-oxononanoic acid (3.94 g, 12.61 mmol, 1 equivalent) in DCM (40 mL), EDCI (2.90 g, 15.13 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (1 g, 5.68 mmol, 0.45 equivalents), DIPEA (4.07 g, 31.53 mmol, 5.49 mL, 2.5 equivalents), and DMAP (154.07 mg, 1.26 mmol, 0.1 equivalent) were added under an N2 atmosphere. This mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 90 mL (30 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O9-[[2,2-dimethyl-5-[[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]nonanedioate (2 g, 2.61 mmol, yield 20.73%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.51-5.49(m,2H),5.35-5.32(m,2H),4.10(s,4H),4.06(t,J=12.0Hz,8H),3.74(s,4H),2.40-2.35(m,4H),2.3 3-2.27(m,4H),2.04(t,J=8.0Hz,4H),1.6(t,J=8.0Hz,9H),1.42(s,6H),1.34-1.31(m,20H),0.92-0.88(m,6H).

[0388] Step 3: [ka] O9-[[2,2-dimethyl-5-[[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O1-[(Z)-nona-3-enyl]nonanedioate (6.5 g, 8.50 mmol, 1 equivalent) was dissolved in THF (65 mL) and HCl (3 M, 3.12 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with saturated NaHCO3 (100 mL) and extracted with 300 mL (100 mL x 3) of ELISA. The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). Compound O9-[2,2-bis(hydroxymethyl)-3-[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]nonanedioate (2.5 g, 3.45 mmol, yield 40.59%) was obtained as a colorless oil.

[0389] Step 4: [ka] To a solution of O9-[2,2-bis(hydroxymethyl)-3-[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxypropyl]O1-[(Z)-nona-3-enyl]nonanedioate (0.6 g, 827.58 μmol, 1 equivalent) in DCM (6 mL), EDCI (190.38 mg, 993.10 μmol, 1.2 equivalents), 9-[(Z)-nona-3-enoxy]-9-oxo-nonanoic acid (155.14 mg, 496.55 μmol, 0.6 equivalents), DIPEA (267.40 mg, 2.07 mmol, 360.38 μL, 2.5 equivalents) and DMAP (10.11 mg, 82.76 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O9-[2-(hydroxymethyl)-3-[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxy-2-[[9-[(Z)-nona-3-enoxy]-9-oxononanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]nonanedioate (0.3 g, 294.28 μmol, yield 35.56%) was obtained as a colorless oil.

[0390] Step 5: [ka] O9-[2-(hydroxymethyl)-3-[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxy-2-[[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]nonanedioate (1.5g, 1.47 mmol, 1 equivalent) in DCM (20mL) solution with EDCI (338.49mg) 1.77 mmol, 1.2 equivalents, DIPEA (475.42 mg, 3.68 mmol, 640.73 μL, 2.5 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (579.93 mg, 1.77 mmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (21.81 mg, 147.14 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was concentrated under pressure to remove the solvent. The residue was diluted with H2O (30 mL) and extracted with DCM 90 mL (30 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O9-[2,2-bis[[9-[(Z)-nona-3-enoxy]-9-oxo-nonanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]nonanedioate (0.1 g, 75.20 μmol, yield 5.11%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.48(m,3H),5.36-5.33(m,3H),4.76(br s,1H),4.11(s,8H),4.07(t,J=7.0Hz,6H),3.34(br s,2H),2.75-2.55(m,4H),2.38(m,8H),2.30(m,12H),2.04(m,6H),1.90- 1.73(m,6H),1.70-1.58(m,16H),1.38-1.27(m,44H),0.91-0.85(m,12H).

[0391] Example 30 - Compound 23: O8-[2,2-bis[(8-nonoxy-8-oxo-octanoy))oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-nonyloctanedioate Step 1: [ka] To a solution of nonane-1-ol (20 g, 138.64 mmol, 1 equivalent) in DCM (600 mL) and THF (600 mL), octanedioic acid (120.76 g, 693.22 mmol, 5 equivalents), DIPEA (53.76 g, 415.93 mmol, 72.45 mL, 3 equivalents), EDCI (34.55 g, 180.24 mmol, 1.3 equivalents), and DMAP (3.39 g, 27.73 mmol, 0.2 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was poured into H2O (1000 mL) and extracted with DCM (300 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 3 / 1). Compound 8-nonoxy-8-oxo-octanoic acid (33 g, 109.84 mmol, yield 79.23%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.40(br s,1H),4.06(t,J=6.8Hz,2H),2.43-2.20(m,4H),1.73-1.55(m,6H),1.43-1.15(m,16H),0.96-0.81(m,3H).

[0392] Step 2: [ka] 8-nonoxy-8-oxo-octanoic acid (18.94 g, 63.06 mmol, 1 equivalent) and [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (5 g, 28.38 mmol, 0.45 equivalents) were dissolved in DCM (200 mL). DIPEA (20.37 g, 157.64 mmol, 27.46 mL, 2.5 equivalents), EDCI (14.51 g, 75.67 mmol, 1.2 equivalents), and DMAP (770.34 mg, 6.31 mmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was poured into H2O (50 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 10 / 1). Compound O8-[[2,2-dimethyl-5-[(8-nonoxy-8-oxo-octanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-nonyloctanedioate (17 g, 22.94 mmol, yield 72.65%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,4H),4.06(t,J=6.8Hz,4H),3.74(s,4H),2.31(m,8H),1.70-1.55(m,12H),1.42(s,6H),1.39-1.09(m,32H),0.94-0.80(m,6H).

[0393] Step 3: [ka] O8-[[2,2-dimethyl-5-[(8-nonoxy-8-oxo-octanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-nonyloctanedioate (17 g, 22.94 mmol, 1 equivalent) was dissolved in THF (170 mL) and HCl (3 M, 9.18 mL, 1.2 equivalents) was added at 0°C under N2. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with saturated NaHCO3 (100 mL) and extracted with HCl 300 mL (100 mL x 3). The combined organic layers were concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 3 / 1). Compound O8-[2,2-bis(hydroxymethyl)-3-(8-nonoxy-8-oxo-octanoyl)oxypropyl]O1-nonyloctanedioate (8.7 g, 12.41 mmol, yield 54.10%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.13(s,4H),4.05(t,J=6.8Hz,4H),3.58(br s,4H),2.86(br s,2H),2.40-2.16(m,8H),1.71-1.50(m,12H),1.41-1.13(m,32H),0.87(t,J=6.8Hz,6H).

[0394] Step 4: [ka] 8-nonoxy-8-oxo-octanoic acid (2.24 g, 7.45 mmol, 0.6 equivalents) and O8-[2,2-bis(hydroxymethyl)-3-(8-nonoxy-8-oxo-octanoyl)oxypropyl]O1-nonyloctanedioate (8.7 g, 12.41 mmol, 1 equivalent) were dissolved in DCM (87 mL) and DIPEA (4.01 g, 31.03 mmol, 5.40 mL, 2.5 equivalents), EDCI (2.86 g, 14.89 mmol, 1.2 equivalents), and DMAP (151.62 mg, 1.24 mmol, 0.1 equivalent) were added. The reaction mixture was stirred at 20 °C for 12 hours under an N2 atmosphere. The reaction mixture was poured into H2O (100 mL) and extracted with DCM (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1 to 5 / 1). Compound O8-[2-(hydroxymethyl)-3-(8-nonoxy-8-oxo-octanoyl)oxy-2-[(8-nonoxy-8-oxooctanoyl)oxymethyl]propyl]O1-nonyloctanedioate (3.4 g, 3.46 mmol, yield 27.85%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.10(s,6H),4.04(t,J=6.8Hz,6H),3.49(br d,J=6.2Hz,2H),2.65(br t,J=6.6Hz,1H),2.30(m,12H),1.52-1.68(m,18H),1.11-1.39(m,48H),0.77-0.90(m,9H),

[0395] Step 5: [ka] To a solution of O8-[2-(hydroxymethyl)-3-(8-nonoxy-8-oxo-octanoyl)oxy-2-[(8-nonoxy-8-oxo-octanoyl)-oxymethyl]propyl]O1-nonyloctanedioate (800 mg, 813.51 μmol, 1 equivalent) and 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (534.39 mg, 1.63 mmol, 2 equivalents) in DCM (8 mL), DIPEA (262.85 mg, 2.03 mmol, 354.25 μL, 2.5 equivalents), EDCI (187.14 mg, 976.21 μmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (12.06 mg, 81.35 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was poured into H2O (20 mL) and extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by Lamb chromatography (SiO2, dichloromethane / methanol = 50 / 1 to 5 / 1). Compound O8-[2,2-bis[(8-nonoxy-8-oxo-octanoy))oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-nonyloctanedioate (110 mg, 81.64 μmol, yield 10.06%, purity 96.02%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.19(br s,1H),4.75(br s,1H),4.11(s,8H),4.06(t,J=6.8Hz,6H),3.29(br d,J=5.4Hz,2H),3.29(br d,J=5.4Hz,2H),2.59(br t,J=5.8Hz,2H),2.52(br s,4H),2.42-2.34(m,2H),2.33-2.19(m,12H),1.89(br d,J=6.8Hz,1H),1.78(br s,5H),1.70(br s,2H),1.61(br d,J=3.2Hz,18H),1.43-1.22(m,56H),0.92-0.84(m,12H).

[0396] Example 31 - Compound 24: O7-[2,2-bis[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-nonylheptanediate Step 1: [ka] Nonane-1-ol (20 g, 138.64 mmol, 1 equivalent) was mixed with DCM (200 mL) and THF (200 mL) to which EDCI (34.55 g, 180.24 mmol, 1.3 equivalents), heptanediol (111.03 g, 693.22 mmol, 5 equivalents), DMAP (1.69 g, 13.86 mmol, 0.1 equivalent), and DIPEA (44.80 g, 346.61 mmol, 60.37 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1). Compound 7-nonoxy-7-oxo-heptanoic acid (30 g, 104.75 mmol, yield 75.55%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.78-10.36(br s,1H),4.06(t,J=6.8Hz,2H),2.49-2.24(m,4H),1.65(m,6H),1.50-1.49(m,14H),0.99-0.80(m,3H).

[0397] Step 2: [ka] To a solution of 7-nonoxy-7-oxoheptanoic acid (14.45 g, 50.44 mmol, 1 equivalent) in DCM (145 mL), EDCI (11.60 g, 60.53 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (4 g, 22.70 mmol, 0.45 equivalents), DMAP (616.27 mg, 5.04 mmol, 0.1 equivalent), and DIPEA (16.30 g, 126.11 mmol, 21.97 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (300 mL) and extracted with 800 mL (200 mL x 4) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1). Compound O7-[[2,2-dimethyl-5-[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-nonylheptanedioate (7.5 g, 10.52 mmol, yield 20.85%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,4H),4.06(t,J=6.8Hz,4H),3.74(s,4H),2.32(m,8H),1.72-1.55(m,12H),1.42(s,6H),1.40-1.09(m,28H),0.88(t,J=6.8Hz,6H).

[0398] Step 3: [ka] O7-[[2,2-dimethyl-5-[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-nonylheptanedioate (7 g, 9.82 mmol, 1 equivalent) was dissolved in THF (70 mL) and HCl (3 M, 3.60 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 7 hours under an N2 atmosphere. The reaction mixture was diluted with saturated NaHCO3 (50 mL) and extracted with 100 mL (50 mL x 2) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O7-[2,2-bis(hydroxymethyl)-3-(7-nonoxy-7-oxo-heptanoyl)oxypropyl]O1-nonylheptanedioate (2.5 g, 3.72 mmol, yield 37.84%) was obtained as a colorless oil.

[0399] Step 4: [ka] To a 25 mL solution of O7-[2,2-bis(hydroxymethyl)-3-(7-nonoxy-7-oxo-heptanoyl)oxypropyl]O1-nonylheptanediote (2.5 g, 3.72 mmol, 1 equivalent) in DCM (25 mL), EDCI (854.63 mg, 4.46 mmol, 1.2 equivalents), 7-nonoxy-7-oxo-heptanoic acid (638.42 mg, 2.23 mmol, 0.6 equivalents), DMAP (45.39 mg, 371.51 μmol, 0.1 equivalent), and DIPEA (1.20 g, 9.29 mmol, 1.62 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM 200 mL (50 mL x 4). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound O7-[2-(hydroxymethyl)-3-(7-nonoxy-7-oxo-heptanoyl)oxy-2-[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]propyl]O1-nonylheptanedioate (1.2 g, 1.27 mmol, yield 34.31%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,6H),4.05(t,J=13.6Hz,6H),3.51(d,J=6.4Hz,2H),2.66(t,J=6.8Hz,1H) ,2.36-2.29(m,12H),1.68-1.60(m,18H),1.39-1.27(m,42H),0.92-0.87(m,9H).

[0400] Step 5: [ka] O7-[2-(hydroxymethyl)-3-(7-nonoxy-7-oxo-heptanoyl)oxy-2-[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]propyl]O1-nonylheptanedioate (1.2g, 1.27 mmol, 1 equivalent) in DCM (12 mL) solution with EDCI (293.26 mg, 1.53 mmol, 1.2 equivalents) 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (251.22 mg, 764.89 μmol, 0.6 equivalents), 4-pyrrolidine-1-ylpyridine (18.89 mg, 127.48 μmol, 0.1 equivalent), and DIPEA (411.90 mg, 3.19 mmol, 555.12 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 50 mL (25 mL x 2) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O7-[2,2-bis[(7-nonoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-nonylheptanedioate (120 mg, 91.78 μmol, yield 7.20%, purity 95.74%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.20(s,1H),4.76-4.75(s,1H),4.11(s,8H),4.06(t,J=6.8Hz,6H),3.30-3.29(s,2H),2.57(d,J=30.8Hz,6H),2.4 0-2.29(m,14H),1.91-1.79(m,6H),1.68-1.56(m,22H),1.52-1.45(m,2H),1.39-1.28(m,46H),0.93-0.87(m,12H).

[0401] Example 32 - Compound 25: O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] To a solution of NaOH (14.77 g, 369.21 mmol, 1.05 equivalents) in H2O (250 mL), 5-butyltetrahydrofuran-2-one (50 g, 351.63 mmol, 50.97 mL, 1 equivalent) was added under an N2 atmosphere. The reaction mixture was stirred at 100 °C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. Compound 4-hydroxyoctanoyloxysodium (45 g, 246.99 mmol, yield 70.24%) was obtained as a white solid and was used directly in the next step.

[0402] Step 2: [ka] BnBr (8.45 g, 49.40 mmol, 5.87 mL, 1 equivalent) was added dropwise to a DMSO (90 mL) solution of 4-hydroxyoctanoyloxysodium (9 g, 49.40 mmol, 1 equivalent) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 5 minutes. The reaction mixture was diluted with saturated NaCl (100 mL) and extracted with siRNA (200 mL) (100 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound benzyl 4-hydroxyoctanoate (2.67 g, crude) was obtained as a pale yellow oil and was immediately used in the next step.

[0403] Step 3: [ka] To a solution of benzyl 4-hydroxyoctanoate (8 g, 31.96 mmol, 1 equivalent) in DCM (201 mL), (4-nitrophenyl)carbonochloride (12.88 g, 63.91 mmol, 2 equivalents) and Py. (5.06 g, 63.91 mmol, 5.16 mL, 2 equivalents) were slowly added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 1 hour under an N2 atmosphere. The reaction mixture was diluted with petroleum ether (100 mL), filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound benzyl 4-(4-nitrophenoxy)carbonyloxyoctanoate (5 g, 12.04 mmol, yield 37.66%) was obtained as a colorless oil.

[0404] Step 4: [ka] To a solution of benzyl 4-(4-nitrophenoxy)carbonyloxyoctanoate (5 g, 12.04 mmol, 1 equivalent) in DCM (77 mL), DIPEA (4.67 g, 36.11 mmol, 6.29 mL, 3 equivalents), 2-pyrrolidine-1-ylethaneamine (2.75 g, 24.07 mmol, 2 equivalents), and DMAP (147.03 mg, 1.20 mmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (100 mL) and extracted with 360 mL (120 mL x 3) of DCM. The combined organic layer was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). The compound benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoate (2 g, 5.12 mmol, yield 42.55%) was obtained as a colorless oil.

[0405] Step 5: [ka] To a suspension of Pd / C (6.54 g, 6.15 mmol, 10% purity, 1.2 equivalents) in THF (40 mL), benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoate (2 g, 5.12 mmol, 1 equivalent) was added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under H2 (15 Psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 50 / 1~3 / 1). Compound 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoic acid (0.5 g, 1.66 mmol, yield 32.50%) was obtained as a pale yellow oil. 1 H NMR(400MHz,CDCl3)δ ppm 6.56(d,J=2.8Hz,1H),4.75-4.68(m,1H),3.65-3.57(m,1H),3.22-3.16(m,1H),3.07-2.83(m,6H),2.34-2.2 3(m,2H),2.00-1.95(m,5H),1.83-1.76(m,1H),1.62-1.46(m,2H),1.33-1.29(m,4H),0.86(t,J=6.4Hz,3H).

[0406] Step 6: [ka] O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (0.5g, 511.59μmol, 1 equivalent) in a 5mL solution of DCM with EDCI (117.69m g (613.91 μmol, 1.2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoic acid (230.52 mg, 767.38 μmol, 1.5 equivalents), DIPEA (165.30 mg, 1.28 mmol, 222.77 μL, 2.5 equivalents), and 4-pyrrolidine-1-ylpyridine (7.58 mg, 51.16 μmol, 0.1 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (30 mL) and extracted with DCM 60 mL (20 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (105 mg, 83.35 μmol, yield 16.29%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.47(m,3H),5.37-5.33(m,3H),4.76(s,1H),4.11(s,8H),4.07(t,J=7.2Hz,6H),3.39(s,2H),2.78(s,4H),2.39-2.35(m,8H),2.33 -2.27(m,12H),2.07-1.99(m,6H),1.89-1.77(m,6H),1.67-1.57(m,14H),1.50-1.46(m,1H),1.39-1.23(m,36H),0.89(t,J=6.8Hz,12H).

[0407] Example 33 - Compound 26: O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate [ka] To a 10 mL solution of O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl] in DCM (196.77 mg, 1.03 mmol, 1.2 equivalents), DIPEA (276.38 mg, 2.14 mmol, 372.47 μL, 2.5 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoic acid (308.34 mg, 1.03 mmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (12.68 mg, 85.54 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)octanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.12 g, 98.55 μmol, yield 11.52%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.52-5.50(m,3H),5.35-5.33(m,3H),4.75(s,1H),4.11(s,8H),4.07(t,J=7.2Hz,6H),3.32(d,J=5.2Hz,2H),2.60(d,J=25.2Hz,6H),2 .40-2.29(m,21H),2.07-2.01(m,6H),2.07-1.99(m,6H),1.80(s,6H),1.68-1.59(m,14H),1.39-1.29(m,28H),0.90(t,J=6.8Hz,12H).

[0408] Example 34 - Compound 27: O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)dodecanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] 5-Octyltetrahydrofuran-2-one (50 g, 252.14 mmol, 1 equivalent) was dissolved in H2O (500 mL) and NaOH (10.59 g, 264.75 mmol, 1.05 equivalent) was added under an N2 atmosphere. The reaction mixture was stirred at 100 °C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. Compound 4-hydroxydodecanoyloxysodium (50 g, crude) was obtained as a white solid and was used directly in the next step.

[0409] Step 2: [ka] BnBr (7.18 g, 41.96 mmol, 4.98 mL, 1 equivalent) was added to a DMSO (100 mL) solution of 4-hydroxydodecanoyloxysodium (10 g, 41.96 mmol, 1 equivalent) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 0.5 hours. The reaction mixture was diluted with saturated NaCl (100 mL) and extracted with ELISA (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. Compound benzyl 4-hydroxydecanoate (30 g, crude) was obtained as a pale yellow oil and was immediately used in the next step.

[0410] Step 3: [ka] To a solution of benzyl 4-hydroxydodecanoate (10 g, 32.63 mmol, 1 equivalent) in DCM (100 mL), Py (5.16 g, 65.27 mmol, 5.27 mL, 2 equivalents) and (4-nitrophenyl)carbonochloride (7.89 g, 39.16 mmol, 1.2 equivalents) were added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 1 hour under an N2 atmosphere. The reaction mixture was diluted with petroleum ether (100 mL), filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). Compound benzyl 4-(4-nitrophenoxy)carbonyl oxide dodecanoate (9 g, 19.09 mmol, yield 81.82%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 8.28-8.24(m,2H),7.41-7.33(m,7H),5.14(s,2H),4.92-4.85(m,1H),2.52(t,J=7.6Hz,2H),2.11(m,1 H),2.04-1.94(m,1H),1.82-1.69(m,1H),1.68-1.60(m,1H),1.48-1.25(m,12H),0.89(t,J=6.8Hz,3H).

[0411] Step 4: [ka] 90 mL of DCM (Dihydrocarbonate) solution of benzyl 4-(4-nitrophenoxy)carbonyl oxide dodecanoate (9 g, 19.09 mmol, 1 equivalent) and 2-pyrrolidine-1-ylethaneamine (4.36 g, 38.17 mmol, 2 equivalents) was mixed with DIEA (7.40 g, 57.26 mmol, 9.97 mL, 3 equivalents) and DMAP (233.17 mg, 1.91 mmol, 0.1 equivalent) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25 °C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 200 mL (100 mL x 2) of DCM. The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). The compound benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy) dodecanoate (3 g, 6.72 mmol, yield 35.21%) was obtained as a yellow oil. 1 H NMR(400MHz,CDCl3)δ ppm 10.55(br s,1H),7.49-7.29(m,5H),5.29(br d,J=6.2Hz,1H),5.11(s,2H),4.74(br s,1H),3.42-3.16(m,2H),2.78-2.56(m,6H),2.38(br t,J=7.6Hz,2H),2.01-1.89(m,1H),1.83(br s,4H),1.55-1.37(m,2H),1.36-1.22(m,12H),0.87(t,J=6.8Hz,3H).

[0412] Step 5: [ka] Benzyl 4-(2-pyrrolidine-1-ylethylcarbamoyloxy) dodecanoate (3 g, 6.72 mmol, 1 equivalent) was added to a suspension of Pd / C (714.83 mg, 671.71 μmol, purity 10%, 0.1 equivalent) in THF (180 mL) under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under H2 (15 Psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 50 / 1~3 / 1). Compound 4-(2-pyrrolidine-1-ylethylcarbamoyloxy) dodecanoic acid (1.8 g, 5.05 mmol, yield 90.00%) was obtained as a yellow oil. 1 H NMR(400MHz,CDCl3)δ ppm 8.87(br s,1H),6.43(br d,J=3.38Hz,1H),4.73(br d,J=2.50Hz,1H),3.63-3.60(m,1H),3.24-3.13(m,1H),3.10-2.89(m,5H),2.85-2.83(m,1H),2.40-2.17(m,2H),2. 11-1.89(m,5H),1.87-1.70(m,1H),1.69-1.54(m,1H),1.51-1.46(m,1H),1.38-1.22(m,12H),0.87(t,J=6.82Hz,3H)

[0413] Step 6: [ka] O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanediolate (800 mg, 818.54 μmol, 1 equivalent) and 4-(2-pyrrolidine-1-ylethylcarbamate) To a 10 mL solution of yloxydodecanoic acid (437.71 mg, 1.23 mmol, 1.5 equivalents) in DCM, DIEA (264.48 mg, 2.05 mmol, 356.44 μL, 2.5 equivalents), EDCI (188.30 mg, 982.25 μmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (12.13 mg, 81.85 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was diluted with H2O (30 mL) and extracted with 90 mL (30 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)dodecanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (300 mg, 183.06 μmol, yield 22.30%, purity 80.29%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,3H),5.37-5.33(m,3H),4.75(br s,1H),4.18-4.02(m,14H),3.35(br d,J=4.8Hz,2H),2.81-2.48(m,6H),2.37(m,8H),2.04(m,6H),2.30(m,12H),1 .94-1.78(m,6H),1.71-1.58(m,12H),1.43-1.10(m,46H),0.91-0.84(m,12H).

[0414] Example 35 - Compound 28: O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)dodecanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanediate [ka] O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.8g, 855.37μmol, 1 equivalent) in DCM (110mL) solution with EDCI (1 96.77 mg, 1.03 mmol, 1.2 equivalents) of 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)dodecanoic acid (274.44 mg, 769.83 μmol, 0.9 equivalents), DMAP (261.24 mg, 2.14 mmol, 2.5 equivalents), and 4-pyrrolidine-1-ylpyridine (12.68 mg, 85.54 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 20°C for 12 hours under an N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (200 mL) and extracted with DCM 300 mL (100 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)dodecanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.12 g, 94.21 μmol, yield 11.01%) was obtained as a colorless oil. 1H NMR(400MHz,CDCl3)δ ppm 5.49-5.52(m,3H),5.35-5.32(m,3H),4.67(s,1H),4.03(s,8H),3.99(t,J=7.2Hz,6H),3.70-3.26(m,2H),2.60-2.46(m ,6H),2.33-2.21(m,20H),1.99-1.91(m,6H),1.75(s,6H),1.58-1.53(m,14H),1.31-1.19(m,36H),0.84-0.79(m,12H).

[0415] Example 36 - Compound 29: O8-[2-[4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoyloxymethyl]-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] A mixture of benzyl 4-(4-nitrophenoxy)carbonyl oxydecanoate (9 g, 20.29 mmol, 1 equivalent), DIPEA (7.87 g, 60.88 mmol, 10.60 mL, 3 equivalents), and DMAP (247.92 mg, 2.03 mmol, 0.1 equivalents) in DCM (90 mL) was mixed with N'-ethyl-N'-methylethane-1,2-diamine (5.18 g, 50.73 mmol, 2.5 equivalents) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25 °C for 12 hours. The reaction mixture was diluted with H2O (40 mL) and extracted with 120 mL of ethyl acetate (40 mL x 3). The combined organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). The compound benzyl 4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoate (5 g, 12.30 mmol, yield 60.60%) was obtained as a colorless oil. 1H NMR(400MHz,MeOD)δ ppm 7.31-7.28(m,5H),4.86(s,2H),4.77-4.71(m,1H),3.23(t,J=6.8Hz,2H),2.60-2.47(m,4H ),2.42(t,J=7.4Hz,2H),2.29(s,3H),1.92(m,1H),1.80(m,1H),1.64-1.48(m,2H),1.30(br d,J=5.2Hz,8H),1.08(t,J=7.2Hz,3H),0.95-0.82(m,3H).

[0416] Step 2: [ka] Benzyl 4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoate (5 g, 12.30 mmol, 1 equivalent) was added to a suspension of Pd / C (300 mg, 10% purity) in THF (100 mL) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under H2 (15 Psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 ~ DCM:MeOH = 0:1). Compound 4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoic acid (2.2 g, 6.95 mmol, yield 56.53%) was obtained as a pale yellow oil. 1 H NMR(400MHz,MeOD)δ ppm 4.84-4.70(m,1H),3.31-3.26(m,1H),3.23-3.01(m,5H),2.75(s,3H),2.27-2.20(m,1H),2.14-1.98(m ,2H),1.80-1.72(m,1H),1.61-1.57(m,1H),1.48-1.44(m,1H),1.30-1.27(m,10H),0.88-0.84(m,3H).

[0417] Step 3: [ka] O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (1.1g, 1.13 mmol, 1 equivalent), 4-[2-[ethyl(methyl)amino]ethylcarbamoyl A mixture of oxydecanoic acid (427.37 mg, 1.35 mmol, 1.2 equivalents), DIPEA (363.66 mg, 2.81 mmol, 490.10 μL, 2.5 equivalents), and EDCI (258.91 mg, 1.35 mmol, 1.2 equivalents) in 20 mL of DCM was mixed with 4-pyrrolidine-1-ylpyridine (16.68 mg, 112.55 μmol, 0.1 equivalent) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). O8-[2-[4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoyloxymethyl]-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxooctanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (100 mg, 78.38 μmol, yield 6.96%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.49(m,3H),5.35-5.33(m,3H),4.75(s,1H),4.11(s,8H),4.06(t,J=3.4Hz,6H),3.29(s,2H),2.53-2.37(m,4H),2.40-2.33(m,8H), 2.31-2.78(m,14H),2.07-2.01(m,6H),1.64-1.57(m,14H),1.50-1.4 0(m,2H),1.36-1.27(m,40H),1.10-1.05(m,3H),0.91-0.86(m,12H).

[0418] Example 37 - Compound 30: O7-[2-[4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoyloxymethyl]-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanediate [ka] O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.8g, 855.37μmol, 1 equivalent), 4-[2-[ethyl(methyl)amino]ethylcarbamate A mixture of yloxydecanoic acid (324.80 mg, 1.03 mmol, 1.2 equivalents), DIPEA (276.38 mg, 2.14 mmol, 372.47 μL, 2.5 equivalents), and EDCI (196.77 mg, 1.03 mmol, 1.2 equivalents) in 10 mL of DCM was mixed with 4-pyrrolidine-1-ylpyridine (12.68 mg, 85.54 μmol, 0.1 equivalent) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). O7-[2-[4-[2-[ethyl(methyl)amino]ethylcarbamoyloxy]decanoyloxymethyl]-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (0.1 g, 81.06 μmol, yield 9.48%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.50(m,3H),5.36-5.33(m,3H),4.75(s,1H),4.11(s,8H),4.06(t,J=3.4Hz,6H),3.37(s,2 H),2.65-2.37(m,3H),2.40-2.34(m,22H),1.68-1.60(m,22H),1.38-1.24(m,32H),0.89(m,12H).

[0419] Example 38 - Compound 31: O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate Step 1: [ka] To a solution of benzyl 4-(4-nitrophenoxy)carbonyl oxydecanoate (2 g, 4.51 mmol, 1 equivalent) in ACN (28 mL), Py. (713.43 mg, 9.02 mmol, 727.99 μL, 2 equivalents), 3-pyrrolidine-1-ylpropan-1-ol (873.98 mg, 6.76 mmol, 1.5 equivalents) and DMAP (55.09 mg, 450.97 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with H2O (100 mL) and extracted with 400 mL (100 mL x 4) of siRNA. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound benzyl 4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoate (1 g, 2.31 mmol, yield 51.14%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 7.37-7.27(m,5H),5.12(s,2H),4.75-4.72(m,1H),4.20-4.16(m,2H),2.59-2.55(m,2H),2.64-2.49(m,6H),1 .99(m,1H),1.95-1.56(m,3H),1.89-1.81(m,4H),1.70-1.48(m,2H),1.38-1.25(m,8H),0.88(t,J=6.6Hz,3H).

[0420] Step 2: [ka] Benzyl 4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoate (1 g, 2.31 mmol, 1 equivalent) was added to a suspension of Pd / C (333.33 mg, 313.22 μmol, 10% purity, 1.36 e-1 equivalent) in THF (20 mL) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under H2 (15 Psi). The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 100 / 1~0 / 1). Compound 4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoic acid (0.4 g, 1.16 mmol, yield 50.50%) was obtained as a pale yellow oil. 1 H NMR(400MHz,CDCl3)δ ppm 8.24-8.17(m,1H),4.84-4.82(m,1H),4.34-4.32(m,1H),4.02(m,1H),3.32-3.05(m,3H),2.98-2.87(m,2H),2.26-2.73(m,1H), 2.39-2.20(m,2H),2.15-2.03(m,1H),2.01-1.83(m,7H),1.78-1.56(m,1H),1.52(m,1H),1.38-1.07(m,8H),0.95-0.76(m,3H).

[0421] Step 3: [ka] O8-[2-(hydroxymethyl)-3-[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxy-2-[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]octanedioate (1g, 1.02 mmol, 1 equivalent) in DCM (1.13 mL) with EDCI (235.37 mg) added. 1.23 mmol, 1.2 equivalents, 4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoic acid (316.27 mg, 920.86 μmol, 0.9 equivalents), 4-pyrrolidine-1-ylpyridine (15.16 mg, 102.32 μmol, 0.1 equivalent), and DIPEA (330.60 mg, 2.56 mmol, 445.55 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O8-[2,2-bis[[8-[(Z)-nona-3-enoxy]-8-oxo-octanoyl]oxymethyl]-3-[4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]octanedioate (0.1 g, 76.76 μmol, yield 7.50%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.54-5.48(m,3H),5.37-5.31(m,3H),4.72-4.70(m,1H),4.21-4.20(m,2H),4.11(d,J=4.0Hz,8H),4.06(t,J=7.2Hz,6H),2.63(s,4H),2. 40-2.35(m,8H),2.33-2.28(m,12H),2.05-2.01(m,10H),1.88-1.84(m,4H),1.64-1.59(m,16H),1.34-1.28(m,38H),0.91-0.87(m,12H).

[0422] Example 39 - Compound 32: [ka] O7-[2-(hydroxymethyl)-3-[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxy-2-[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (1.5g, 1.60 mmol, 1 equivalent) in DCM (20 mL) solution with EDCI (3 68.94 mg, 1.92 mmol, 1.2 equivalents) of 4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoic acid (495.76 mg, 1.44 mmol, 0.9 equivalents), DMAP (19.59 mg, 160.38 μmol, 0.1 equivalent), and DIPEA (518.20 mg, 4.01 mmol, 698.39 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with DCM 200 (100 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound O7-[2,2-bis[[7-[(Z)-nona-3-enoxy]-7-oxo-heptanoyl]oxymethyl]-3-[4-(3-pyrrolidine-1-ylpropoxycarbonyloxy)decanoyloxy]propyl]O1-[(Z)-nona-3-enyl]heptanedioate (100 mg, 78.88 μmol, yield 4.92%, purity 99.45%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.52-5.50(m,3H),5.35-5.33(m,3H),4.74-4.70(m,1H),4.22(t,J=6Hz,2H),4.11(s,8H),4.06(t,J=7.2Hz,6H),3.04-2.88(m,4H),2.38 (t,J=6.4Hz,7H),2.35-2.27(m,12H),2.20-2.18(m,2H),2.07-2.01(m,10H),1.66-1.60(m,20H),1.37-1.28(m,30H),0.91-0.87(m,12H).

[0423] Example 40 - Compound 33: O8-[2,2-bis[(8-octoxy-8-oxo-octanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-octyloctanedioate Step 1: [ka] To a mixture of octan-1-ol (20 g, 153.58 mmol, 24.27 mL, 1 equivalent), DIPEA (59.55 g, 460.73 mmol, 80.25 mL, 3 equivalents), DMAP (3.75 g, 30.72 mmol, 0.2 equivalents), and EDCI (38.27 g, 199.65 mmol, 1.3 equivalents) in DCM (800 mL) and THF (800 mL), octaneoic acid (133.76 g, 767.89 mmol, 5 equivalents) was added under an N2 atmosphere, and the reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 8-octoxy-8-oxo-octanoic acid (25 g, 87.29 mmol, yield 56.84%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.23(br s,1H),4.06(t,J=6.8Hz,2H),2.42-2.22(m,4H),1.73-1.55(m,6H),1.46-1.26(m,14H),0.89(t,J=6.8Hz,3H).

[0424] Step 2: [ka] A mixture of 8-octoxy-8-oxo-octanoic acid (14.45 g, 50.44 mmol, 1 equivalent), DIPEA (16.30 g, 126.11 mmol, 21.97 mL, 2.5 equivalents), EDCI (11.60 g, 60.53 mmol, 1.2 equivalents), and DMAP (616.27 mg, 5.04 mmol, 0.1 equivalent) in 200 mL of DCM was mixed with [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (4 g, 22.70 mmol, 0.45 equivalents) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[[2,2-dimethyl-5-[(8-octoxy-8-oxo-octanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-octyloctanedioate (6.5 g, 9.12 mmol, yield 18.07%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.10(s,4H),4.06(t,J=6.8Hz,4H),3.74(s,4H),2.30(m,8H),1.69-1.59(m,12H),1.42(s,6H),1.36-1.24(m,28H),0.94-0.83(m,6H).

[0425] Step 3: [ka] O8-[[2,2-dimethyl-5-[(8-octoxy-8-oxo-octanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-octyloctanedioate (6.5 g, 9.12 mmol, 1 equivalent) was dissolved in THF (65 mL) and HCl (3 M, 3.75 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with NaHCO3 aqueous solution (200 mL) and extracted with ELISA 250 mL (50 mL x 5). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). Compound O8-[2,2-bis(hydroxymethyl)-3-(8-octoxy-8-oxo-octanoyl)oxypropyl]O1-octyloctanedioate (2.5 g, 3.72 mmol, yield 40.75%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.15(s,4H),3.98(t,J=6.8Hz,4H),3.57(s,4H),2.86(br s,2H),2.34-2.16(m,8H),1.59-1.46(m,12H),1.31-1.17(m,28H),0.90-0.72(m,6H).

[0426] Step 4: [ka] To a mixture of O8-[2,2-bis(hydroxymethyl)-3-(8-octoxy-8-oxo-octanoyl)oxypropyl]O1-octyloctanedioate (1.3 g, 1.93 mmol, 1 equivalent), EDCI (444.41 mg, 2.32 mmol, 1.2 equivalents), DMAP (23.60 mg, 193.19 μmol, 0.1 equivalent), and DIPEA (624.20 mg, 4.83 mmol, 841.24 μL, 2.5 equivalents) in 150 mL of DCM, 8-octoxy-8-oxo-octanoic acid (331.98 mg, 1.16 mmol, 0.6 equivalents) was added under an N2 atmosphere, and the reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2-(hydroxymethyl)-3-(8-octoxy-8-oxo-octanoyl)oxy-2-[(8-octoxy-8-oxo-octanoyl)oxymethyl]propyl]O1-octyloctanedioate (500 mg, 531.17 μmol, yield 27.50%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,6H),4.06(t,J=6.8Hz,6H),3.50(d,J=6.8Hz,2H),2.60(t,J=6.8Hz,1 H),2.38-2.27(m,12H),1.62(m,18H),1.36-1.23(m,42H),0.93-0.82(m,9H).

[0427] Step 5: [ka] O8-[2-(hydroxymethyl)-3-(8-octoxy-8-oxo-octanoyl)oxy-2-[(8-octoxy-8-oxo-octanoyl)oxymethyl]propyl]O1-octyloctanedioate (1.1g, 1.17 mmol, 1 equivalent), DIPEA (377.58mg, 2.92 mmol, 508.86μL, 2.5 equivalents), EDCI (268.82mg) To a mixture of (1.40 mmol, 1.2 equivalents) and 4-pyrrolidine-1-ylpyridine (17.32 mg, 116.86 μmol, 0.1 equivalents) in DCM (15 mL), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (498.96 mg, 1.52 mmol, 1.3 equivalents) was added under an N2 atmosphere, and the reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2,2-bis[(8-octoxy-8-oxo-octanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-octyloctanedioate (150 mg, 119.83 μmol, yield 10.25%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.18(s,1H),4.75(s,1H),4.14-4.11(m,8H),4.05(t,J=3.4Hz,2H),3.29(d,J=2.6Hz,2H),2.60-2.52(m,6H),2.3 3-2.29(m,14H),1.90(d,J=3.0Hz,1H),1.18(s,4H),1.62-1.61(m,20H),1.35-1.25(m,52H),0.90-0.86(m,12H).

[0428] Example 41 - Compound 34: O7-[2,2-bis[(7-octoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-octylheptanedioate Step 1: [ka] A solution of heptanediol (122.99 g, 767.89 mmol, 5 equivalents) in DCM (615 mL) and THF (615 mL) was added to EDCI (38.27 g, 199.65 mmol, 1.3 equivalents), DMAP (1.88 g, 15.36 mmol, 0.1 equivalent), DIPEA (49.62 g, 383.94 mmol, 66.88 mL, 2.5 equivalents), and octan-1-ol (20 g, 153.58 mmol, 24.27 mL, 1 equivalent), and stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (300 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 7-octoxy-7-oxo-heptanoic acid (30 g, 110.14 mmol, yield 71.72%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.02(br s,1H),4.06(t,J=6.8Hz,2H),2.43-2.24(m,4H),1.74-1.56(m,6H),1.45-1.16(m,12H),0.94-0.80(m,3H).

[0429] Step 2: [ka] To a solution of 7-octoxy-7-oxo-heptanoic acid (13.74 g, 50.44 mmol, 1 equivalent) in DCM (130 mL), EDCI (11.60 g, 60.53 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (4 g, 22.70 mmol, 0.45 equivalents), DMAP (616.27 mg, 5.04 mmol, 0.1 equivalent), and DIPEA (16.30 g, 126.11 mmol, 21.97 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O7-[[2,2-dimethyl-5-[(7-octoxy-7-oxo-heptanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-octylheptanedioate (7g, 10.22 mmol, yield 20.26%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,4H),4.06(t,J=6.8Hz,4H),3.74(s,4H),2.32(m,8H),1.70-1.59(m,12H),1.42(s,6H),1.40-1.25(m,24H),0.95-0.81(m,6H).

[0430] Step 3: [ka] O7-[[2,2-dimethyl-5-[(7-octoxy-7-oxo-heptanoyl)oxymethyl]-1,3-dioxan-5-yl]methyl]O1-octylheptanedioate (7g, 10.22 mmol, 1 equivalent) was dissolved in THF (70 mL) and HCl (3M, 3.75 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with NaHCO3 aqueous solution (200 mL) and extracted with HCl 250 mL (50 mL x 5). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O7-[2,2-bis(hydroxymethyl)-3-(7-octoxy-7-oxo-heptanoyl)oxypropyl]O1-octylheptanedioate (2.5 g, 3.88 mmol, yield 37.93%) was obtained as a colorless oil.

[0431] Step 4: [ka] To a 10 mL solution of O7-[2,2-bis(hydroxymethyl)-3-(7-octoxy-7-oxo-heptanoyl)oxypropyl]O1-octylheptanedioate (1 g, 1.55 mmol, 1 equivalent) in DCM (10 mL), EDCI (356.72 mg, 1.86 mmol, 1.2 equivalents), 7-octoxy-7-oxo-heptanoic acid (253.43 mg, 930.41 μmol, 0.6 equivalents), DMAP (18.94 mg, 155.07 μmol, 0.1 equivalent), and DIPEA (501.04 mg, 3.88 mmol, 675.26 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound O7-[2-(hydroxymethyl)-3-(7-octoxy-7-oxo-heptanoyl)oxy-2-[(7-octoxy-7-oxo-heptanoyl)oxymethyl]propyl]O1-octylheptanedioate (0.4 g, 444.82 μmol, yield 28.69%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.16(s,6H),4.06(t,J=6.4Hz,6H),3.52(d,J=6.8Hz,2H),2.66(t,J=6.8Hz,1H),2 .36-2.29(m,12H),1.69-1.59(m,19H),1.40-1.27(m,34H),0.89(t,J=13.6Hz,9H).

[0432] Step 5: [ka] To a 5 mL solution of O7-[2-(hydroxymethyl)-3-(7-octoxy-7-oxo-heptanoyl)oxy-2-[(7-octoxy-7-oxo-heptanoyl)oxymethyl]propyl]O1-octylheptanedioate (0.4 g, 444.82 μmol, 1 equivalent) in DCM (5 mL), EDCI (102.33 mg, 533.79 μmol, 1.2 equivalents), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (131.49 mg, 400.34 μmol, 0.9 equivalents), DIPEA (143.72 mg, 1.11 mmol, 193.70 μL, 2.5 equivalents) and DMAP (5.43 mg, 44.48 μmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with 50 mL (25 mL x 2) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O7-[2,2-bis[(7-octoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-octylheptanedioate was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.19(s,1H),4.76(d,J=2.0Hz,1H),4.12(s,8H),4.06(t,J=6.8Hz,6H),3.30(d,J=5.2Hz,1H),2.61(t,J=12Hz,2H) ,2.52(s,4H),2.40-2.29(m,14H),1.95-1.78(m,6H),1.68-1.60(m,22H),1.39-1.28(m,42H),0.90-0.87(m,12H).

[0433] Example 42 - Compound 35: O8-[2,2-bis[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-decyloctanedioate Step 1: [ka] A mixture of decane-1-ol (20 g, 126.36 mmol, 24.13 mL, 1 equivalent), DMAP (1.54 g, 12.64 mmol, 0.1 equivalent), EDCI (29.07 g, 151.63 mmol, 1.2 equivalents), and DIPEA (40.83 g, 315.90 mmol, 55.02 mL, 2.5 equivalents) in DCM (65 mL) and THF (65 mL) was mixed with octanedioic acid (110.05 g, 631.79 mmol, 5 equivalents) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25 °C for 12 hours. The reaction mixture was diluted with H2O (600 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 8-decoxy-8-oxo-octanoic acid (25 g, 79.50 mmol, yield 62.92%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.71-10.55(m,1H),4.06(t,J=3.2Hz,2H),2.37-2.28(m,4H),1.69-1.58(m,6H),1.43-1.27(m,18H),0.89(t,J=3.4Hz,3H).

[0434] Step 2: [ka] [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (2g, 11.35 mmol, 0.45 equivalents) was added to a mixture of 8-decoxy-8-oxo-octanoic acid (7.93g, 25.22 mmol, 1 equivalent), DIPEA (8.15g, 63.06 mmol, 10.98 mL, 2.5 equivalents), EDCI (5.80g, 30.27 mmol, 1.2 equivalents), and DMAP (616.27 mg, 5.04 mmol, 0.2 equivalents) in DCM (300 mL) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[[5-[(8-decoxy-8-oxo-octanoyl)oxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methyl]O1-decyloctanedioate (4 g, 5.20 mmol, yield 20.62%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.10(s,4H),4.05(t,J=6.8Hz,4H),3.74(s,4H),2.30-2.90(m,8H),1.6 3-1.60(m,12H),1.42(s,6H),1.22-1.36(m,36H),0.88(t,J=6.8Hz,6H).

[0435] Step 3: [ka] O8-[[5-[(8-decoxy-8-oxo-octanoyl)oxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methyl]O1-decyloctanedioate (4 g, 5.20 mmol, 1 equivalent) was dissolved in THF (40 mL) and HCl (2 M, 2.86 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). Compound O8-[2-[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-hydroxy-2-(hydroxymethyl)propyl]O1-decyloctanedioate (1.5 g, 2.06 mmol, yield 39.56%) was obtained as a colorless oil.

[0436] Step 4: [ka] 8-decoxy-8-oxo-octanoic acid (388.20 mg, 1.23 mmol, 0.6 equivalents) was added to a mixture in 15 mL of DCM containing O8-[2-[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-hydroxy-2-(hydroxymethyl)propyl]O1-decyloctanedioate (1.5 g, 2.06 mmol, 1 equivalent), EDCI (473.32 mg, 2.47 mmol, 1.2 equivalents), DMAP (25.14 mg, 205.75 μmol, 0.1 equivalent), and DIPEA (664.80 mg, 5.14 mmol, 895.96 μL, 2.5 equivalents). The mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2,2-bis[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-hydroxypropyl]O1-decyloctanedioate (800 mg, 780.12 μmol, yield 37.92%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,6H),4.05(t,J=6.8Hz,6H),3.50(d,J=6.8Hz,1H),3.56-3.43(m,1H),2.61(t,J=6 .6Hz,1H),2.37-2.27(m,12H),1.67-1.58(m,18H),1.37-1.20(m,54H),0.97-0.73(m,9H).

[0437] Step 5: [ka] To a 20 mL solution of O8-[2,2-bis[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-hydroxy-propyl]O1-decyloctanedioate (1.5 g, 1.46 mmol, 1 equivalent) in DCM (20 mL), 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (576.51 mg, 1.76 mmol, 1.2 equivalents), DIPEA (472.62 mg, 3.66 mmol, 636.95 μL, 2.5 equivalents), 4-pyrrolidine-1-ylpyridine (216.78 mg, 1.46 mmol, 1 equivalent), and EDCI (336.49 mg, 1.76 mmol, 1.2 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL of ethyl acetate (20 mL x 3). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O8-[2,2-bis[(8-decoxy-8-oxo-octanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-decyloctanedioate (110 mg, 82.34 μmol, yield 5.63%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.18(br s,1H),4.75(br s,1H),4.14-4.09(m,8H),4.06(t,J=6.8Hz,6H),3.29(d,J=5.2Hz,2H),2.59(t,J=5.9Hz,2H),2.52(br s,4H),2.40-2.34(m,2H),2.33-2.27(m,12H),1.94-1.84(m,1H),1.78(br s,4H),1.61(m,18H),1.44-1.11(m,66H),0.96-0.81(m,12H).

[0438] Example 43 - Compound 36: O7-[2,2-bis[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-decylheptanediate Step 1: [ka] A mixture of decane-1-ol (20 g, 126.36 mmol, 24.13 mL, 1 equivalent), EDCI (29.07 g, 151.63 mmol, 1.2 equivalents), DMAP (1.54 g, 12.64 mmol, 0.1 equivalent), and DIPEA (40.83 g, 315.90 mmol, 55.02 mL, 2.5 equivalents) in DCM (600 mL) and THF (600 mL) was mixed with heptanediic acid (101.19 g, 631.79 mmol, 5 equivalents) under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 7-decoxy-7-oxo-heptanoic acid (30 g, 99.86 mmol, yield 79.03%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 11.27(br s,1H),4.06(t,J=6.8Hz,2H),2.45-2.29(m,4H),1.71-1.56(m,6H),1.45-1.24(m,16H),0.96-0.83(m,3H)

[0439] Step 2: [ka] A mixture of [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (3g, 17.03 mmol, 0.45 equivalents), DMAP (462.20 mg, 3.78 mmol, 0.1 equivalent), EDCI (8.70 g, 45.40 mmol, 1.2 equivalents), and DIPEA (12.22 g, 94.58 mmol, 16.47 mL, 2.5 equivalents) in DCM (150 mL) was mixed with 7-decoxy-7-oxoheptanoic acid (11.37 g, 37.83 mmol, 1 equivalent) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (200 mL) and extracted with 600 mL (200 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[[5-[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methyl]O1-decylheptanedioate (3.5 g, 4.72 mmol, yield 12.48%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,4H),4.06(t,J=6.8Hz,4H),3.74(s,4H),2.32(m,8H),1.73-1.59(m,12H),1.42(s,6H),1.39-1.23(m,32H),0.90-0.86(m,6H).

[0440] Step 3: [ka] A mixture of O7-[[5-[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-2,2-dimethyl-1,3-dioxan-5-yl]methyl]O1-decylheptanedioate (3.5 g, 4.72 mmol, 1 equivalent) in THF (35 mL) was mixed with HCl (3 M, 1.73 mL, 1.1 equivalents) at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with aqueous NaHCO3 solution (200 mL) and extracted with 600 mL of ethyl acetate (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1~0 / 1). Compound O7-[2-[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-hydroxy-2-(hydroxymethyl)propyl]O1-decylheptanedioate (1.2 g, 1.71 mmol, yield 36.25%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.15(s,4H),4.06(t,J=6.8Hz,4H),3.59(br d,J=4.2Hz,4H),2.78(br s,2H),2.45-2.25(m,8H),1.69-1.59(m,12H),1.40-1.24(m,32H),0.97-0.82(m,6H).

[0441] Step 4: [ka] 7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-hydroxy-2-(hydroxymethyl)propyl]O1-decylheptanedioate (1 g, 1.43 mmol, 1 equivalent), DIPEA (460.94 mg, 3.57 mmol, 621.21 μL, 2.5 equivalents), EDCI (328.17 mg, 1.71 mmol, 1.2 equivalents), and DMAP (17.43 mg, 142.66 μmol, 0.1 equivalent) were mixed in DCM (15 mL) to which 7-decoxy-7-oxo-heptanoic acid (257.15 mg, 855.94 μmol, 0.6 equivalents) was added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25 °C for 12 hours. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2,2-bis[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-hydroxy-propyl]O1-decylheptanedioate (0.4 g, 406.75 μmol, yield 28.51%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.11(s,6H),4.05(t,J=6.8Hz,6H),3.51(br d,J=6.4Hz,2H),2.68(t,J=6.6Hz,1H),2.32(m,12H),1.68-1.59(m,18H),1.40-1.24(m,48H),0.88(t,J=6.6Hz,9H)

[0442] Step 5: [ka] 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (280.55 mg, 854.18 μmol, 1.2 equivalents) was added to a mixture of O7-[2,2-bis[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-hydroxy-propyl]O1-decylheptanedioate (0.7 g, 711.82 μmol, 1 equivalent), DIPEA (229.99 mg, 1.78 mmol, 309.96 μL, 2.5 equivalents), EDCI (163.75 mg, 854.18 μmol, 1.2 equivalents), and 4-pyrrolidine-1-ylpyridine (10.55 mg, 71.18 μmol, 0.1 equivalent) in DCM (10 mL) under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (20 mL) and extracted with 60 mL (20 mL x 3) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O7-[2,2-bis[(7-decoxy-7-oxo-heptanoyl)oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O1-decylheptanedioate (130 mg, 100.48 μmol, yield 14.12%, purity 100%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.21(br s,1H),4.75(br s,1H),4.11(s,8H),4.06(t,J=6.8Hz,6H),3.30(br d,J=4.8Hz,2H),2.60(br s,2H),2.53(br s,4H),2.42-2.36(m,2H),2.31(d,J=7.4Hz,10H),1.90(m,1H),1.78(br s,6H),1.71-1.59(m,20H),1.46-1.24(m,58H),0.98-0.82(m,12H).

[0443] Example 44 - Compound 37: O1-[2,2-bis[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O8-(1-methyloctyl)octanedioate Step 1: [ka] To a 600 mL solution of octanedioic acid (60.38 g, 346.61 mmol, 5 equivalents) in DCM, EDCI (15.95 g, 83.19 mmol, 1.2 equivalents), DMAP (846.89 mg, 6.93 mmol, 0.1 equivalent), and DIPEA (22.40 g, 173.31 mmol, 30.19 mL, 2.5 equivalents) were added under an N2 atmosphere. Then, nonan-2-ol (10 g, 69.32 mmol, 1 equivalent) in 100 mL of DCM was added to the above reaction mixture, and the reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with 150 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 8-(1-methyloctoxy)-8-oxo-octanoic acid (6 g, 19.97 mmol, yield 28.81%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 10.42(br s,1H),4.80-5.00(m,1H),2.36(t,J=7.4Hz,2H),2.28(t,J=7.6Hz,2H),1.71-1.41(m,5H),1.49 -1.41(m,1H),1.40-1.33(m,4H),1.33-1.22(m,10H),1.20(d,J=6.2Hz,3H),0.96-0.80(m,3H).

[0444] Step 2: [ka] To a solution of 8-(1-methyloctoxy)-8-oxo-octanoic acid (15.16 g, 50.44 mmol, 1 equivalent) in DCM (200 mL), EDCI (11.60 g, 60.53 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (4 g, 22.70 mmol, 0.45 equivalents), DMAP (616.27 mg, 5.04 mmol, 0.1 equivalent), and DIPEA (16.30 g, 126.11 mmol, 21.97 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 800 mL (200 mL x 4) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[[2,2-dimethyl-5-[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O8-(1-methyloctyl)octanedioate (8g, 10.80 mmol, yield 21.40%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.87(m,2H),4.11(s,4H),3.74(s,4H),2.30(m,8H),1.71-1.56(m,10H),1.48(m,2H),1 .42(s,6H),1.39-1.31(m,10H),1.31-1.25(m,18H),1.20(d,J=6.2Hz,6H),0.93-0.83(m,6H).

[0445] Step 3: [ka] O1-[[2,2-dimethyl-5-[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O8-(1-methyloctyl)octanedioate (7g, 9.45 mmol, 1 equivalent) was dissolved in THF (70 mL) and HCl (3M, 3.46 mL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with NaHCO3 aqueous solution (200 mL) and extracted with HCl 600 mL (200 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis(hydroxymethyl)-3-[8-(1-methyloctoxy)-8-oxo-octanoyl]oxypropyl]O8-(1-methyloctyl)octanedioate (4g, 5.71 mmol, yield 60.41%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,2H),4.15(s,4H),3.59(s,4H),2.78(br s,2H),2.36(t,J=7.6Hz,4H),2.28(t,J=7.4Hz,4H),1.63(m,10H),1.52-1 .42(m,2H),1.37-1.23(m,28H),1.20(d,J=6.2Hz,6H),0.96-0.80(m,6H).

[0446] Step 4: [ka] To a solution of O1-[2,2-bis(hydroxymethyl)-3-[8-(1-methyloctoxy)-8-oxo-octanoyl]oxypropyl]O8-(1-methyloctyl)octanedioate (3 g, 4.28 mmol, 1 equivalent) in DCM (37 mL), EDCI (984.51 mg, 5.14 mmol, 1.2 equivalents), 8-(1-methyloctoxy)-8-oxo-octanoic acid (771.46 mg, 2.57 mmol, 0.6 equivalents), DMAP (52.28 mg, 427.97 μmol, 0.1 equivalent), and DIPEA (1.38 g, 10.70 mmol, 1.86 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 400 mL (100 mL x 4) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1). Compound O1-[2-(hydroxymethyl)-3-[8-(1-methyloctoxy)-8-oxo-octanoyl]oxy-2-[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]propyl]O8-(1-methyloctyl)octanedioate (1.5 g, 1.53 mmol, yield 35.64%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.86(m,3H),4.12(s,6H),3.51(d,J=9.6Hz,2H),2.61(t,J=6.8Hz,1H),2.35-2.26(m,12H),1.64-1.61(m, 12H),1.49-1.43(m,4H),1.36-1.32(m,14H),1.30-1.28(m,30H),1.20(t,J=6.4Hz,9H),0.88(t,J=13.6Hz,9H).

[0447] Step 5: [ka] O1-[2-(hydroxymethyl)-3-[8-(1-methyloctoxy)-8-oxo-octanoyl]oxy-2-[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]propyl]O8-(1-methyloctyl)octanedioate (1.8g, 1.83 mmol, 1 equivalent) in DCM (19 mL) solution with EDCI (421.07 mg, 2. 20 mmol, 1.2 equivalents) of 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (541.07 mg, 1.65 mmol, 0.9 equivalents), 4-pyrrolidine-1-ylpyridine (27.13 mg, 183.04 μmol, 0.1 equivalent), and DIPEA (591.41 mg, 4.58 mmol, 797.05 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 400 ml (100 mL x 4) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound O1-[2,2-bis[[8-(1-methyloctoxy)-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O8-(1-methyloctyl)octanedioate (0.1 g, 61.83 μmol, yield 3.38%, purity 80%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.40-5.35(m,1H),4.94-4.74(m,3H),4.75(s,1H),4.11-4.09(s,8H),3.37(s,2H),2.72-2.60(s,6H),2.40-2.25(m,14H),1 .86-1.77(m,6H),1.64-1.55(m,18H),1.49-1.45(m,4H),1.35-1.27(m,48H),1.20(d,J=6.4Hz,9H),0.88(t,J=13.6Hz,12H).

[0448] Example 45 - Compound 38: O1-[2,2-bis[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O7-(1-methyloctyl)heptanedioate Step 1: [ka] To a solution of nonan-2-ol (20 g, 138.64 mmol, 1 equivalent) in DCM (1310 mL), EDCI (34.55 g, 180.24 mmol, 1.3 equivalents), heptaneoic acid (111.03 g, 693.22 mmol, 5 equivalents), DMAP (1.69 g, 13.86 mmol, 0.1 equivalent), and DIPEA (44.80 g, 346.61 mmol, 60.37 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 7-(1-methyloctoxy)-7-oxo-heptanoic acid (30 g, 104.75 mmol, yield 75.55%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.93-4.88(m,1H),2.37(t,J=7.6Hz,2H),2.29(t,J=7.6Hz,2H),1.74-1.61(m,4H),1.61-1. 52(m,1H),1.52-1.34(m,3H),1.33-1.23(m,10H),1.20(d,J=6.2Hz,3H),0.96-0.82(m,3H).

[0449] Step 2: [ka] To a solution of 7-(1-methyloctoxy)-7-oxo-heptanoic acid (3.61 g, 12.61 mmol, 1 equivalent) in DCM (46 mL), EDCI (2.90 g, 15.13 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (1 g, 5.68 mmol, 0.45 equivalents), DMAP (154.07 mg, 1.26 mmol, 0.1 equivalent), and DIPEA (4.07 g, 31.53 mmol, 5.49 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound O1-[[2,2-dimethyl-5-[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O7-(1-methyloctyl)heptanedioate (1.5 g, 2.10 mmol, yield 16.68%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,2H),4.11(s,4H),3.75(s,4H),2.37-2.13(m,8H),1.69-1.60(m,8H),1.47(br d,J=8.0Hz,2H),1.56(s,2H),1.42(s,6H),1.39-1.22(m,24H),1.20(d,J=6.2Hz,6H),0.98-0.71(m,6H).

[0450] Step 3: [ka] O1-[[2,2-dimethyl-5-[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O7-(1-methyloctyl)heptanedioate (1.5 g, 2.10 mmol, 1 equivalent) was dissolved in THF (15 mL) and HCl (3 M, 771.40 μL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with NaHCO3 aqueous solution (50 mL) and extracted with EtOAC 100 mL (50 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis(hydroxymethyl)-3-[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxypropyl]O7-(1-methyloctyl)heptanedioate (0.6 g, 891.63 μmol, yield 42.38%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.87(m,2H),4.15(s,4H),3.59(s,4H),2.37(t,J=7.2Hz,4H),2.29(t,J=7.6Hz,4H),1.7 0-1.55(m,12H),1.50-1.32(m,8H),1.28(s,18H)1.20(d,J=6.0Hz,6H),0.89(t,J=6.4Hz,6H).

[0451] Step 4: [ka] To a solution of O1-[2,2-bis(hydroxymethyl)-3-[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxypropyl]O7-(1-methyloctyl)heptanedioate (0.6 g, 891.63 μmol, 1 equivalent) in DCM (1 mL), EDCI (205.11 mg, 1.07 mmol, 1.2 equivalents), 7-(1-methyloctoxy)-7-oxo-heptanoic acid (153.22 mg, 534.98 μmol, 0.6 equivalents), DMAP (10.89 mg, 89.16 μmol, 0.1 equivalent), and DIPEA (288.09 mg, 2.23 mmol, 388.26 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (50 mL) and extracted with 100 mL (50 mL x 2) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1). Compound O1-[2-(hydroxymethyl)-3-[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxy-2-[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]propyl]O7-(1-methyloctyl)heptanedioate (0.3 g, 318.70 μmol, yield 35.74%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.87(m,3H),4.11(s,6H),3.51(d,J=6.4Hz,2H),2.69(t,J=13.6Hz,1H),2.36-2.26(m,12H),1.69-1. 62(m,12H),1.58-1.44(m,6H),1.40-1.32(m,8H),1.27(s,28H),1.20(d,J=6Hz,9H),0.88(t,J=13.6Hz,9H).

[0452] Step 5: [ka] O1-[2-(hydroxymethyl)-3-[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxy-2-[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]propyl]O7-(1-methyloctyl)heptanedioate (0.9g, 956.11μmol, 1 equivalent) in DCM (10mL) solution with EDCI (219.94mg, 1 0.15 mmol, 1.2 equivalents) of 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (282.63 mg, 860.50 μmol, 0.9 equivalents), 4-pyrrolidine-1-ylpyridine (14.17 mg, 95.61 μmol, 0.1 equivalent), and DIPEA (308.93 mg, 2.39 mmol, 416.34 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis[[7-(1-methyloctoxy)-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O7-(1-methyloctyl)heptanedioate (0.1 g, 79.82 μmol, yield 8.35%, purity 99.92%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.35-5.32(s,1H),4.92-4.87(m,3H),4.75(s,1H),4.11(s,8H),3.34(d,J=4.0Hz,2H),2.65(d,J=18.4Hz,6H),2.39 -2.26(m,14H),1.83(s,6H),1.67-1.46(m,22H),1.39-1.27(m,42H),1.20(d,J=6.4Hz,9H),0.88(t,J=6.4Hz,12H).

[0453] Example 46 - Compound 39: O1-[2,2-bis[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O8-(1-methylnonyl)octanedioate Step 1: [ka] To a solution of decane-2-ol (20 g, 126.36 mmol, 1 equivalent) in DCM (1300 mL), EDCI (31.49 g, 164.27 mmol, 1.3 equivalents), octanedioic acid (110.05 g, 631.79 mmol, 5 equivalents), DMAP (1.54 g, 12.64 mmol, 0.1 equivalent), and DIPEA (40.83 g, 315.90 mmol, 55.02 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 8-(1-methylnonoxy)-8-oxo-octanoic acid (30 g, 95.40 mmol, yield 75.50%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.93-4.88(m,1H),2.36(t,J=7.6Hz,2H),2.28(t,J=7.6Hz,2H),1.74-1.52(m,6H) ,1.51-1.42(m,1H),1.38-1.22(m,16H),1.20(d,J=6.2Hz,3H),0.94-0.81(m,3H).

[0454] Step 2: [ka] To a 50 mL solution of 8-(1-methylnonoxy)-8-oxo-octanoic acid (3.97 g, 12.61 mmol, 1 equivalent) in DCM, EDCI (2.90 g, 15.13 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (1 g, 5.68 mmol, 0.45 equivalents), DMAP (154.07 mg, 1.26 mmol, 0.1 equivalent), and DIPEA (4.07 g, 31.53 mmol, 5.49 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 150 mL (50 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O1-[[2,2-dimethyl-5-[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O8-(1-methylnonyl)octanedioate (1.5 g, 1.95 mmol, yield 15.47%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,2H),4.11(s,4H),3.75(s,4H),2.30(m,8H),1.68-1.56(m,10H),1.48 (m,2H),1.42(s,6H),1.36-1.23(m,32H),1.20(d,J=6.2Hz,6H),0.96-0.81(m,6H),

[0455] Step 3: [ka] O1-[[2,2-dimethyl-5-[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O8-(1-methylnonyl)octanedioate (1.5 g, 1.95 mmol, 1 equivalent) was dissolved in THF (15 mL) and HCl (3 M, 715.12 μL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with saturated NaHCO3 (100 mL) and extracted with ELISA 200 mL (50 mL x 4). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis(hydroxymethyl)-3-[8-(1-methylnonoxy)-8-oxo-octanoyl]oxypropyl]O8-(1-methylnonyl)octanediate (0.8 g, 1.10 mmol, yield 56.26%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,2H),4.15(s,4H),3.58(br s,4H),2.79(br s,2H),2.35(t,J=7.4Hz,4H),2.27(t,J=7.4Hz,4H),1.66-1.53(m,10H),1.4 9-1.41(m,2H),1.39-1.2(m,32H),1.20(d,J=6.25Hz,6),0.97-0.82(m,6H).

[0456] Step 4: [ka] To a 10 mL solution of O1-[2,2-bis(hydroxymethyl)-3-[8-(1-methylnonoxy)-8-oxo-octanoyl]oxypropyl]O8-(1-methylnonyl)octanediate (0.8 g, 1.10 mmol, 1 equivalent) in DCM (10 mL), EDCI (252.43 mg, 1.32 mmol, 1.2 equivalents), 8-(1-methylnonoxy)-8-oxo-octanoic acid (207.04 mg, 658.41 μmol, 0.6 equivalents), DMAP (13.41 mg, 109.73 μmol, 0.1 equivalent), and DIPEA (354.56 mg, 2.74 mmol, 477.84 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O1-[2-(hydroxymethyl)-3-[8-(1-methylnonoxy)-8-oxo-octanoyl]oxy-2-[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]propyl]O8-(1-methylnonyl)octanedioate (0.3 g, 292.55 μmol, yield 26.66%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.87(m,3H),4.12(s,6H),3.51(d,J=2.8Hz,2H),2.62(s,1H),2.33(t,J=7.6Hz,6H),2.27(t,J=7.2Hz,6H),1. 64-1.61(m,10H),1.49-1.42(m,6H),1.36-1.32(m,14H),1.27(s,34H),1.20(d,J=6.4Hz,9H),0.88(t,J=6.4Hz,9H).

[0457] Step 5: [ka] O1-[2-(hydroxymethyl)-3-[8-(1-methylnonoxy)-8-oxo-octanoyl]oxy-2-[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]propyl]O8-(1-methylnonyl)octanediate (1 g, 975.16 μmol, 1 equivalent) in a DCM (13 mL) solution with EDCI (224.33 mg, 1.17 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (288.26 mg, 877.64 μmol, 0.9 equivalents), 4-pyrrolidine-1-ylpyridine (14.45 mg, 97.52 μmol, 0.1 equivalent), and DIPEA (315.08 mg, 2.44 mmol, 424.64 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with 100 mL (25 mL x 4) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound O1-[2,2-bis[[8-(1-methylnonoxy)-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O8-(1-methylnonyl)octanedioate (0.1 g, 74.86 μmol, yield 7.68%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.31-5.29(s,1H),4.93-4.86(m,3H),4.75(s,1H),4.11(s,8H),3.33(d,J=4Hz,2H),2.61(d,J=25.6Hz,6H),2.40-2.25(m,14H),1.90 -1.81(m,6H),1.64-1.55(m,18H),1.49-1.42(m,4H),1.35-1.32(m,14H),1.27(s,40H),1.20(d,J=6.4Hz,9H),0.88(t,J=6.4Hz,12H).

[0458] Example 47 - Compound 40: O1-[2,2-bis[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O7-(1-methylnonyl)heptanediate Step 1: [ka] To a solution of decane-2-ol (20 g, 126.36 mmol, 1 equivalent) in DCM (1210 mL), EDCI (29.07 g, 151.63 mmol, 1.2 equivalents), heptanediol (101.19 g, 631.79 mmol, 5 equivalents), DMAP (1.54 g, 12.64 mmol, 0.1 equivalent), and DIPEA (40.83 g, 315.90 mmol, 55.02 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (500 mL) and extracted with 1500 mL (500 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Compound 7-(1-methylnonoxy)-7-oxo-heptanoic acid (30 g, 99.86 mmol, yield 79.03%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,1H),2.36(t,J=7.47Hz,2H),2.29(t,J=7.47Hz,2H),1.74-1.60(m,4H),1.59-1.36(m,4H),1.27(br s,12H),1.20(d,J=6.27Hz,3H),0.93-0.82(m,3H).

[0459] Step 2: [ka] To a 50 mL solution of 7-(1-methylnonoxy)-7-oxoheptanoic acid (3.79 g, 12.61 mmol, 1 equivalent) in DCM (50 mL), EDCI (2.90 g, 15.13 mmol, 1.2 equivalents), [5-(hydroxymethyl)-2,2-dimethyl-1,3-dioxan-5-yl]methanol (1 g, 5.68 mmol, 0.45 equivalents), DMAP (154.07 mg, 1.26 mmol, 0.1 equivalent), and DIPEA (4.07 g, 31.53 mmol, 5.49 mL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25 °C for 12 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with 300 mL (100 mL x 3) of DCM. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[[2,2-dimethyl-5-[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O7-(1-methylnonyl)heptanedioate (1.5 g, 2.02 mmol, yield 16.05%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.88(m,2H),4.11(s,4H),3.74(s,4H),2.42-2.21(m,8H),1.71-1.57(m,10H),1.47(br d,J=8.0Hz,2H),1.42(s,6H),1.38-1.21(m,28H),1.20(d,J=6.2Hz,6H),0.97-0.77(m,6H).

[0460] Step 3: [ka] O1-[[2,2-dimethyl-5-[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]-1,3-dioxan-5-yl]methyl]O7-(1-methylnonyl)heptanedioate (1.5 g, 2.02 mmol, 1 equivalent) was dissolved in THF (15 mL) and HCl (3 M, 742.20 μL, 1.1 equivalents) was added at 0°C under an N2 atmosphere. The reaction mixture was stirred at 25°C for 2 hours under an N2 atmosphere. The reaction mixture was diluted with saturated NaHCO3 (100 mL) and extracted with 200 mL (100 mL x 2) of ethyl acetate. The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis(hydroxymethyl)-3-[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxypropyl]O7-(1-methylnonyl)heptanedioate (0.8 g, 1.14 mmol, yield 56.38%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.92-4.87(m,2H),4.15(s,4H),3.59(br d,J=4.50Hz,4H),2.80(br s,2H),2.37(t,J=7.44Hz,4H),2.29(t,J=7.38Hz,4H),1.69-1.60(m,10H),1.48-1.34(m,6H),1.27(br s,24H),1.20(d,J=6.25Hz,6H),0.89(t,J=6.75Hz,6H).

[0461] Step 4: [ka] To a 10 mL solution of O1-[2,2-bis(hydroxymethyl)-3-[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxypropyl]O7-(1-methylnonyl)heptanedioate (0.8 g, 1.14 mmol, 1 equivalent) in DCM (10 mL), EDCI (262.54 mg, 1.37 mmol, 1.2 equivalents), 7-(1-methylnonoxy)-7-oxo-heptanoic acid (205.72 mg, 684.76 μmol, 0.6 equivalents), DMAP (13.94 mg, 114.13 μmol, 0.1 equivalent), and DIPEA (368.75 mg, 2.85 mmol, 496.97 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred at 25 °C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (100 mL) and extracted with 400 mL (100 mL x 4) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 1 / 1). Compound O1-[2-(hydroxymethyl)-3-[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxy-2-[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]propyl]O7-(1-methylnonyl)heptanedioate (0.3 g, 305.06 μmol, yield 26.73%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 4.93-4.86(m,3H),4.11(s,6H),3.52(d,J=4.8Hz,2H),2.68(s,1H),2.34(t,J=7.6Hz,6H),2.90(t,J=7.6Hz,6 H),1.68-1.60(m,14H),1.49-1.42(m,4H),1.40-1.27(m,42H),1.20(d,J=6.0Hz,9H),0.88(t,J=13.6Hz,9H).

[0462] Step 5: [ka] O1-[2-(hydroxymethyl)-3-[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxy-2-[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]propyl]O7-(1-methylnonyl)heptanedioate (1g, 1.02 mmol, 1 equivalent) in DCM (13 mL) solution with EDCI (233.93 mg, 1.22 mL) 4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoic acid (300.59 mg, 915.19 μmol, 0.9 equivalents), 4-pyrrolidine-1-ylpyridine (15.07 mg, 101.69 μmol, 0.1 equivalent), and DIPEA (328.56 mg, 2.54 mmol, 442.81 μL, 2.5 equivalents) were added under an N2 atmosphere. The reaction mixture was stirred under an N2 atmosphere at 25°C for 12 hours. The reaction mixture was diluted with H2O (50 mL) and extracted with DCM 50 mL (25 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0~0 / 1). Compound O1-[2,2-bis[[7-(1-methylnonoxy)-7-oxo-heptanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O7-(1-methylnonyl)heptanedioate (0.1 g, 77.29 μmol, yield 7.60%) was obtained as a colorless oil. 1 H NMR(400MHz,CDCl3)δ ppm 5.39-5.28(s,1H),4.92-4.87(m,3H),4.75-4.74(s,1H),4.11(s,8H),3.34(d,J=3.6Hz,2H),2.63(d,J=1.6Hz,6H),2.39- 2.26(m,14H),1.91-1.77(m,6H),1.67-1.44(m,26H),1.38-1.27(m,44H),1.20(d,J=6.4Hz,9H),0.88(t,J=13.6Hz,12H).

[0463] Example 48 - Compound 41: O1-[2,2-bis[[8-(1-methyldecoxy)-8-oxo-octanoyl]oxymethyl]-3-[4-(2-pyrrolidine-1-ylethylcarbamoyloxy)decanoyloxy]propyl]O8-(1-methyldecyl)octanedioate Step 1: [ka] To a solution of undecane-2-ol (15 g, 87.05 mmol, 1 equivalent) in DCM (450 mL) and THF (450 mL), EDCI (20.03 g, 104.46 mmol, 1.2 equivalents), octanodioic acid (75.82 g, 435.27 mmol, 5 equivalents), DIPEA (28.13 g, 217.64 mmol, 37.91 mL, 2.5 equivalents), and DMAP (1.06 g, 8.71 mmol, 0.1 equivalent) were added under an N2 atmosphere. The reaction mixture was stirred at 25°C for 12 hours under an N2 atmosphere. The reaction mixture was diluted with H2O (1000 mL) and extracted with 2100 mL (700 mL x 3) of DCM. The combined organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1). Compound 8-(1-methyldecoxy)-8-oxo-octanoic acid (15 g, 4...

Claims

1. The following formula: 【Chemistry 1】 (In the formula, R １ and R １’ each of which is independently (C １ -C ９ alkyl)-R ５ , (C ２ -C ９ alkenyl)-R ５ , (C ２ -C ９ alkynyl)-R ５ or (C １ -C ８ alkoxy)-R ５ and is R １’’ (C １ ~C ９ Alkyl)-R ５ , (C ２ ~C ９ Alkenil)-R ５ , (C ２ ~C ９ Alkinyl)-R ５ , (C １ ~C ８ Alkyl(-R) ５ or R １２ -R １３ And, Each R ５ These are, independently, hydrogen and C １ ~C １２ Alkyl, C ２ ~C １２ Alkenil, C ２ ~C １２ Alkinyl, C ２ ~C １２ Alkoxy, optionally substituted C ３ ~C １２ Cycloalkyl, optionally substituted C ５ ~C ６ Aryl, 1-adamantyl, 2-adamantyl, sterolyl, C(O)O-R ６ OC(O)-R ６ OC(O)O-R ６ CH(R ７ ) R ８ , C(O)O-CH-(R ７ ) R ８ C(O)O-C １ ~C ４ Alkyl-(R ９ ) R １０ OC(O)-C １ ~C ４ Alkyl-(R ９ ) R １０ or OC(O)CH(R ９ ) R １０ And, Each R ６ Independently, C ７ ~C １２ Alkyl, C ７ ~C １２ Alkenil, C ７ ~C １２ Alkinyl, C ７ ~C １２ Alkoxy, optionally substituted C ３ ~C １２ Cycloalkyl, optionally substituted C ５ ~C ６ It is aryl, 1-adamantyl, 2-adamantyl, or sterolyl. Each R ７ and R ８ Independently, C ７ ~C １２ Alkyl, C ７ ~C １２ Alkenil, C ７ ~C １２ Alkinyl or C ７ ~C １２ It is an alkoxy, Each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ It is alkenyl, X １ is O, NH or CHR １４ wherein X ２ is O, NH, or CHR １１ And, R ２ is C １ to C １２ alkyl, C ２ to C １２ alkenyl, C ２ to C １２ alkynyl, C １ to C １２ alkoxy, (C １ to C ４ alkyl)-(C １ to C ４ alkoxy), optionally substituted C ３ to C １２ cycloalkyl, (C １ to C ４ alkyl)-(optionally substituted C ３ to C １２ cycloalkyl), optionally substituted C ３ to C ６ heterocycle, (C １ to C ４ alkyl)-(optionally substituted C ３ to C ６ heterocycle), optionally substituted C ５ to C ６ aryl or (C １ to C ４ alkyl)-(optionally substituted C ５ to C ６ aryl), and R ２’ is hydrogen, C １ ~C １２ Alkyl, alkenyl, or alkynyl, C １ ~C １２ Alkoxy, (C １ ~C ４ (Alkyl)-(C １ ~C ４ It is an alkoxy, R ２ and R ２’ These are combined and optionally substituted C ４ ~C ６ Cycloalkyl, optionally substituted C ３ ~C ６ Cycloalkyl or optionally substituted C ３ ~C ６ It can form a heterogeneous ring, R ３ and R ４ These are, independently, hydrogen and C １ ~C ６ Alkyl, C １ ~C ６ Alkoxy or C １ ~C ６ It is hydroxyalkyl, or R ３ and R ４ They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R １１ is hydrogen or C １ ~C ６ Alkyl or R １１ and R ４ They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R １２ This refers to branched or unbranched C that are joined or optionally substituted. １ ~C ６ Alkyl, C １ ~C ６ Alkenil or C １ ~C ６ It is alkinyl, R １３ C is a hydrogen atom that is optionally substituted with other carbon atoms. ３ ~C １２ Cycloalkyl or optionally substituted C ５ ~C ６ It is Ariel, R １４ is hydrogen or C １ ~C ６ Alkyl or R １４ and R ２ These are joined together and optionally replaced by C ５ ~C ８ Forming a cycloalkyl group, m is 1 to 4, p is 0 to 4, and n is between 1 and 5. A compound of or a pharmaceutically acceptable salt thereof.

2. The following formula: 【Chemistry 2】 (In the formula, R １ and R １’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ , (C ２ ~C ９ Alkenil)-R ５ , (C ２ ~C ９ Alkinyl)-R ５ or (C １ ~C ８ Alkyl(-R) ５ And, R １’’ (C １ ~C ９ Alkyl)-R ５ , (C ２ ~C ９ Alkenil)-R ５ , (C ２ ~C ９ Alkinyl)-R ５ , (C １ ~C ８ Alkyl(-R) ５ or R １２ -R １３ And, Each R ５ These are, independently, hydrogen and C １ ~C １２ Alkyl, C ２ ~C １２ Alkenil, C ２ ~C １２ Alkinyl, C ２ ~C １２ Alkoxy, optionally substituted C ３ ~C １２ Cycloalkyl, optionally substituted C ５ ~C ６ Aryl, 1-adamantyl, 2-adamantyl, sterolyl, C(O)O-R ６ OC(O)-R ６ OC(O)O-R ６ CH(R ７ ) R ８ , C(O)O-CH(R ７ ) R ８ C(O)O-C １ ~C ４ Alkyl-(R ９ ) R １０ OC(O)-C １ ~C ４ Alkyl-(R ９ ) R １０ or OC(O)CH(R ９ ) R １０ And, Each R ６ Independently, C ７ ~C １２ Alkyl, C ７ ~C １２ Alkenil, C ７ ~C １２ Alkinyl, C ７ ~C １２ Alkoxy, optionally substituted C ３ ~C １２ Cycloalkyl, optionally substituted C ５ ~C ６ It is aryl, 1-adamantyl, 2-adamantyl, or sterolyl. Each R ７ and R ８ Independently, C ７ ~C １２ Alkyl, C ７ ~C １２ Alkenil, C ７ ~C １２ Alkinyl or C ７ ~C １２ It is an alkoxy, Each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ It is alkenyl, X １ is O, NH, or CHR １４ And, X ２ is O, NH, or CHR １１ And, R ２ C １ ~C １２ Alkyl, C ２ ~C １２ Alkenyl or C ２ ~C １２ Alkinyl, C １ ~C １２ Alkoxy, (C １ ~C ４ (Alkyl)-(C １ ~C ４ Alkoxy), optionally substituted C ３ ~C １２ Cycloalkyl, (C １ ~C ４ Alkyl)-(Optionally substituted C) ３ ~C １２ Cycloalkyl), optionally substituted C ３ ~C ６ Complex algebra, (C １ ~C ４ Alkyl)-(Optionally substituted C) ３ ~C ６ (Hybrid algebras), C with arbitrary substitution ５ ~C ６ Aryl or (C １ ~C ４ Alkyl)-(Optionally substituted C) ５ ~C ６ (Aryl) R ３ and R ４ These are, independently, hydrogen and C １ ~C ６ Alkyl, C １ ~C ６ Alkoxy or C １ ~C ６ It is hydroxyalkyl, or R ３ and R ４ They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R １２ This refers to branched or unbranched C that are joined or optionally substituted. １ ~C ６ Alkyl, C １ ~C ６ Alkenil or C １ ~C ６ It is alkinyl, R １３ C is a hydrogen atom that is optionally substituted with other carbon atoms. ３ ~C １２ Cycloalkyl or optionally substituted C ５ ~C ６ It is Ariel, R １１ is hydrogen or C １ ~C ６ Alkyl or R １１ and R ４ They bond together to form a heterocyclic ring containing one or more N, O, or S heteroatoms. R １４ is hydrogen or C １ ~C ６ Alkyl or R １４ and R ２ These are joined together and optionally replaced by C ５ ~C ８ Forming a cycloalkyl group, n is 1 to 5, and m is between 1 and 4. A compound of or a pharmaceutically acceptable salt thereof.

3. R １ , R １’ and R １’’ These two are independent of each other, (C １ ~C ９ Alkyl)-R ５ and each R ５ Independently, C ２ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

4. R １ , R １’ and R １’’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ and each R ５ Independently, C ２ ~C １２ The compound according to claim 3, which is an alkenyl.

5. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ C ６ ~C １０ It is a cycloalkyl, and optionally, the R ５’ The compound according to claim 3, wherein is 1-adamantyl or 2-adamantyl.

6. R １ , R １’ and R １’’ One of them is CH(R ７ ) R ８ And each R ７ and R ８ Independently, C ７ ~C １２ The compound according to claim 3, which is an alkoxy.

7. R １ , R １’ and R １’’ These two are independent of each other, (C １ ~C ９ Alkyl)-R ５ And each R ５ It is independently C(O)O-R ６ and each R ６ Independently, C ７ ~C １２ Alkyl or C ７ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

8. R １ , R １’ and R １’’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ And each R ５ It is independently C(O)O-R ６ and each R ６ Independently, C ７ ~C １２ Alkyl or C ７ ~C １２ The compound according to claim 7, which is an alkenyl.

9. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ C ６ ~C １０ It is a cycloalkyl, and optionally, the R ５’ The compound according to claim 6, wherein is 1-adamantyl or 2-adamantyl.

10. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ CH(R ７ ) R ８ And each R ７ and R ８ Independently, C ７ ~C １２ The compound according to claim 7, which is an alkoxy.

11. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ is OC(O)CH(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 7, which is an alkenyl.

12. R １ and R １’ is, (C １ ~C ９ Alkyl)-R ５’ And R ５’ C(O)O-R ６ and each R ６ Independently, C ７ ~C １２ Alkyl or C ７ ~C １２ It is an alkenyl and R １’ R １２ -R １３ And R １２ This refers to branched or unbranched C that are joined or optionally substituted. １ ~C ６ Alkyl, C １ ~C ６ Alkenil or C １ ~C ６ It is an alkynyl and R １３ C is a hydrogen atom that is optionally substituted with other carbon atoms. ３ ~C １２ Cycloalkyl or optionally substituted C ５ ~C ６ The compound according to claim 7, wherein it is an aryl compound.

13. R １２ The compound according to claim 12, wherein the bond is a combination.

14. R １２ C １ ~C ６ Alkyl, optionally C １ The compound according to claim 12, wherein it is alkyl.

15. R １２ This is a branched or unbranched C which is optionally replaced. １ ~C ６ Alkyl, C １ ~C ６ Alkenyl or C １ ~C ６ It is an alkynyl, and optionally R １２ The compound according to claim 12, wherein is selected from pentyl, 1-methylpentyl, 4-methylpentyl, 5,5,5-trifluoropentyl, 4,4,5,5,5-pentafluoropentyl, and penta-4-inyl.

16. R １３ The compound according to any one of claims 12 to 14, wherein is hydrogen.

17. R １３ This is C, which is optionally substituted. ３ ~C １２ It is a cycloalkyl group, and optionally R １３ The compound according to claim 13 or 14, wherein is selected from optionally substituted cyclopropane, optionally substituted cyclobutane, and optionally substituted cyclohexane, for example, 4-pentylcyclohexyl.

18. R １３ This is a condensed C that has been optionally substituted. ３ ~C １２ Cycloalkyl, optionally substituted crosslinked C ３ ~C １２ Cycloalkyl or optionally substituted spiroC ３ ~C １２ The compound described in 17, which is a cycloalkyl compound.

19. R １３ This is C, which is optionally substituted. ５ ~C ６ The compound according to claim 13 or 14, wherein the compound is an aryl, optionally, optionally substituted phenyl, such as 4-pentylphenyl or 3,5-di-tert-butylphenyl.

20. R １３ is a bicyclo[2.2.2]pentane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[2.2.2]pentane, 1-(trifluoromethyl)bicyclo[1.1.1]pentane, or 1-methylbicyclo[1.1.1]pentane.

21. R １３ is a bicyclo[2.1.0]pentane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[2.1.0]pentane.

22. R １３ is a bicyclo[3.1.0]hexane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is 6,6-difluorobicyclo[3.1.0]hexane.

23. R １３ is a bicyclo[2.1.1]hexane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[2.1.1]hexane or 1-fluorobicyclo[2.1.1]hexane.

24. R １３ The compound according to claim 13 or 14, wherein is optionally substituted spiro[2.3]hexane, optionally unsubstituted spiro[2.3]hexane, or 1,1-difluorospiro[2.3]hexane.

25. R １３ The compound according to claim 13 or 14, wherein is optionally substituted 1,1'-bi(cyclohexane).

26. R １３ The compound according to claim 13 or 14, wherein decahydronaphthalene is optionally substituted.

27. R １３ is a bicyclo[2.2.1]heptane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[2.2.1]heptane or 7,7-dimethylbicyclo[2.2.1]heptane.

28. R １３ is a bicyclo[4.1.0]heptane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[4.1.0]heptane or 7,7-difluorobicyclo[4.1.0]heptane.

29. R １３ is a bicyclo[3.2.0]heptane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is an unsubstituted bicyclo[3.2.0]heptane.

30. R １３ is a spiro[3.3]heptane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is an unsubstituted spiro[3.3]heptane or 2,2-difluorospiro[3.3]heptane.

31. R １３ is a bicyclo[2.2.2]octane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[2.2.2]octane or 1-methylbicyclo[2.2.2]octane.

32. R １３ is a bicyclo[3.2.1]octane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted bicyclo[3.2.1]octane or 8-oxabicyclo[3.2.1]octane.

33. R １３ is a spiro[2.5]octane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is unsubstituted spiro[2.5]octane or 1,1-difluorospiro[2.5]octane.

34. R １３ is a bicyclo[3.2.2]nonane that is optionally substituted, and R is optionally substituted. １３ The compound according to claim 13 or 14, wherein is an unsubstituted bicyclo[3.2.2]nonane or 1-fluorobicyclo[3.2.2]nonane.

35. R １３ is an optionally substituted 1-bicyclo[3.3.1]nonane, and optionally R １３ The compound according to claim 13 or 14, wherein is an unsubstituted bicyclo[3.3.1]nonane or 1-methylbicyclo[3.3.1]nonane.

36. R １３ The compound according to claim 13 or 14, wherein is adamantane.

37. R １ , R １’ and R １’’ These two are independent of each other, (C １ ~C ９ Alkyl)-R ５ And each R ５ Independently, OC(O)CH(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

38. R １ , R １’ and R １’’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ And R ５ is OC(O)CH(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 37, which is an alkenyl.

39. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ is C(O)O-C １ ~C ４ Alkyl-(R ９ ) R １０ or OC(O)-C １ ~C ４ Alkyl-(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

40. R １ , R １’ and R １’’ These two are independent of each other, (C １ ~C ９ Alkyl)-R ５ And each R ５ Independently, C(O)O-C １ ~C ４ Alkyl-(R ９ ) R １０ or OC(O)-C １ ~C ４ Alkyl-(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

41. R １ , R １’ and R １’’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ And R ５ is C(O)O-C １ ~C ４ Alkyl-(R ９ ) R １０ or OC(O)-C １ ~C ４ Alkyl-(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 1, wherein the compound is an alkenyl.

42. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ C ６ ~C １０ It is a cycloalkyl group, and optionally R ５’ The compound according to claim 37, wherein is 1-adamantyl or 2-adamantyl.

43. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ C(O)O-R ６ And R ６ C ７ ~C １２ Alkyl or C ７ ~C １２ The compound according to claim 37, which is an alkenyl.

44. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ CH(R ７ ) R ８ And each R ７ and R ８ Independently, C ７ ~C １２ The compound according to claim 37, which is an alkoxy.

45. R １ , R １’ and R １’’ These two are independent of each other, (C １ ~C ９ Alkyl)-R ５ And each R ５ CH(R) ７ ) R ８ And each R ７ and R ８ Independently, C ７ ~C １２ The compound according to claim 1, wherein it is an alkoxy.

46. R １ , R １’ and R １’’ Each of them is independent of (C １ ~C ９ Alkyl)-R ５ And each R ５ CH(R) ７ ) R ８ And each R ７ and R ８ Independently, C ７ ~C １２ The compound according to claim 45, which is an alkoxy.

47. R １ , R １’ and R １’’ One of them is (C １ ~C ９ Alkyl)-R ５’ And R ５’ is OC(O)CH(R ９ ) R １０ And each R ９ and R １０ Independently, C １ ~C １２ Alkyl or C ２ ~C １２ The compound according to claim 42, which is an alkenyl.

48. R ２ C １ ~C １２ Alkyl, C ２ ~C １２ Alkenyl or C ２ ~C １２ The compound according to any one of claims 1 to 47, wherein it is an alkynyl compound.

49. R ２ C ４ ~C ８ A compound according to any one of claims 1 to 47, wherein it is alkyl.

50. R ２ These are methyl, ethyl, propyl, isopropyl, butyl, 1-isobutyl, 2-isobutyl, tert-butyl, C ５ Alkyl, C ６ Alkyl, C ８ Alkyl or C １０ A compound according to any one of claims 1 to 47, wherein it is alkyl.

51. R ２ C １ ~C １２ Alkoxy or (C １ ~C ４ (Alkyl)-(C １ ~C ４ It is an alkoxy, and optionally, R ２ The compound according to any one of claims 1 to 47, wherein is methoxy, ethoxy, methoxymethyl, or ethoxyethyl.

52. R ２ This is C, which is optionally substituted. ３ ~C １２ Cycloalkyl, optionally substituted C ３ ~C ６ A heteroalgebra or a C with arbitrary substitutions ５ ~C ６ It is aryl, and optionally, R ２ The compound according to any one of claims 1 to 47, wherein is optionally substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, for example, 4-pentylcyclohexyl.

53. R ２ is a phenyl that has been optionally substituted, and optionally R ２ The compound according to any one of claims 1 to 47, wherein is 4-pentylphenyl.

54. R ２ (C) is optionally replaced １ ~C ４ Alkyl)-(Optionally substituted C) ３ ~C １２ Cycloalkyl), (C １ ~C ４ Alkyl)-(Optionally substituted C) ３ ~C ６ (Heterogene) or (C １ ~C ４ Alkyl)-(Optionally substituted C) ５ ~C ６ aryl) and optionally, R ２ is, -CH ２ -Cyclopropyl, -(CH ２ ) ２ -Cyclopropyl, -CH ２ -Cyclohexyl, -(CH ２ ) ２ -Cyclohexyl, -(CH ２ ) ２ -(4-pentylcyclohexyl),-CH ２ -Phenyl or -(CH ２ ) ２ - A compound according to any one of claims 1 to 47, wherein the compound is phenyl.

55. R ２’ The compound according to any one of claims 48 to 54, wherein is hydrogen.

56. R ２’ C １ ~C １２ Alkyl, C ２ ~C １２ Alkenyl or C ２ ~C １２ It is an alkynyl, and optionally R ２ The compound according to any one of claims 48 to 54, wherein is methoxy, ethoxy, methoxymethyl, or ethoxyethyl.

57. R ２ and R ２’ These are combined and optionally substituted C ４ ~C ６ Cycloalkyl or C ４ ~C ６ Form a complex algebra, and optionally, R ２ and R ２’ The compound according to any one of claims 1 to 47, wherein the compounds combine to form cyclohexane or pyran.

58. X １ CH ２ The compound according to any one of claims 1 to 57.

59. X １ CHR １４ And R １４ and R ２ These are joined together and optionally replaced by C ５ ~C ８ Form a cycloalkyl group, and optionally, R １４ and R ２ These are joined together and optionally replaced by C ５ Cycloalkyl or optionally substituted C ６ A compound according to any one of claims 1 to 57, which forms a cycloalkyl group.

60. X ２ The compound according to any one of claims 1 to 59, wherein is NH.

61. X ２ The compound according to any one of claims 1 to 59, wherein is O.

62. R ３ and R ４ Each of them is independent of C １ ~C ６ A compound according to any one of claims 1 to 61, wherein it is alkyl.

63. R ３ and R ４ They are bonded together to form a heterocyclic ring containing a nitrogen heteroatom, and optionally, R ３ and R ４ They are bound together to form pyrrolidine, or R ３ and R ４ The compound according to any one of claims 1 to 61, wherein the compound is bonded together with the aforementioned alkyl group to form quinuclidine.

64. The compound according to any one of claims 1 to 63, wherein n is 1, 2, 3, or 4.

65. The compound according to any one of claims 1 to 64, wherein m is 1, 2, 3, or 4.

66. The compound according to any one of claims 1 to 65, wherein p is 0, 1, 2, 3, or 4.

67. R １ , R １’ and R １’’ These two are independent of each other, (C ２ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

68. R １ , R １’ and R １’’ These two are independent of each other, (C ３ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

69. R １ , R １’ and R １’’ These two are independent of each other, (C ４ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

70. R １ , R １’ and R １’’ These two are independent of each other, (C ５ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

71. R １ , R １’ and R １’’ These two are independent of each other, (C ６ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

72. R １ , R １’ and R １’’ These two are independent of each other, (C ７ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

73. R １ , R １’ and R １’’ These two are independent of each other, (C ８ Alkyl)-R ５ The compound according to any one of claims 1 to 66.

74. The following formula: 【Transformation 3】 The compound according to any one of claims 1 to 66, which is the same as or a pharmaceutically acceptable salt thereof.

75. The following formula: 【Chemistry 4】 The compound according to any one of claims 1 to 66, which is the same as or a pharmaceutically acceptable salt thereof.

76. The following formula: 【Transformation 5】 The compound according to any one of claims 1 to 66, which is the same as or a pharmaceutically acceptable salt thereof.

77. The compound according to claim 1, which is one of compounds 1 to 209 or a pharmaceutically acceptable salt thereof.

78. The compound according to claim 1, which is one of compounds 7, 8, 10, 13, 14, 26, 33, 38, 39, 40, 48, 60, 61, 89, 103, or 109, or a pharmaceutically acceptable salt thereof.

79. Lipid nanoparticles comprising the compound described in any one of claims 1 to 78.

80. Lipid nanoparticles comprising the compound, phospholipid, cholesterol, and polyethylene glycol lipid described in any one of claims 1 to 78.

81. Lipid nanoparticles comprising about 20 to 80 mol% of the compound according to any one of claims 1 to 78, about 7.5 to 40 mol% of phospholipids, about 6 to 45 mol% of cholesterol, and about 1 to 4 mol% of PEG lipids.

82. Lipid nanoparticles comprising about 45 to 65 mol% of the compound according to any one of claims 1 to 78, about 10 mol% of phospholipids, about 25 to 45 mol% of cholesterol, and about 1 to 4 mol% of PEG lipids.

83. Lipid nanoparticles comprising approximately 45-50 mol% of the compound according to any one of claims 1 to 78, approximately 10 mol% of phospholipids, approximately 38-42 mol% of cholesterol, and approximately 2-3 mol% of PEG lipids.

84. Lipid nanoparticles according to claim 83, comprising approximately 47.5 mol% of the compound according to any one of claims 1 to 78, approximately 40 mol% of cholesterol, and approximately 2.5 mol% of PEG lipid.

85. Lipid nanoparticles comprising approximately 47.5 to 52.5 mol% of the compound according to any one of claims 1 to 78, approximately 10 mol% of phospholipids, approximately 37 to 40 mol% of cholesterol, and approximately 1 to 2 mol% of PEG lipids.

86. Lipid nanoparticles according to claim 85, comprising about 50 mol% of the compound according to any one of claims 1 to 78, about 38.5 mol% of cholesterol, and about 1.5 mol% of PEG lipid.

87. Lipid nanoparticles comprising approximately 57.5 to 62.5 mol% of the compound according to any one of claims 1 to 78, approximately 10 mol% of phospholipids, approximately 26 to 29 mol% of cholesterol, and approximately 2 to 3 mol% of PEG lipids.

88. Lipid nanoparticles according to claim 87, comprising about 60 mol% of the compound according to any one of claims 1 to 78, about 27.5 mol% of cholesterol, and about 2.5 mol% of PEG lipid.

89. Lipid nanoparticles comprising approximately 45 to 50 mol% of the compound according to any one of claims 1 to 78, approximately 10 mol% of phospholipids, approximately 37.5 to 40.5 mol% of cholesterol, and approximately 3 to 4 mol% of PEG lipids.

90. Lipid nanoparticles according to claim 89, comprising approximately 47.5 mol% of the compound according to any one of claims 1 to 38, approximately 39 mol% of cholesterol, and approximately 3.5 mol% of PEG lipid.

91. Lipid nanoparticles according to any one of claims 80 to 90, further comprising a targeted component.

92. The lipid nanoparticle according to claim 91, wherein the targeted component is a targeted lipid.

93. The lipid nanoparticles according to claim 91, wherein the targeting component is an active targeting component.

94. The lipid nanoparticle according to claim 91, wherein the active targeting component is a protein, peptide, small molecule, antibody, or antigen-binding fragment thereof.

95. The lipid nanoparticle according to any one of claims 79 to 94, further comprising one or more polynucleotides encapsulated within the lipid nanoparticle.

96. The lipid nanoparticles according to claim 95, wherein the one or more polynucleotides include RNA.

97. The lipid nanoparticles according to claim 95, wherein the one or more polynucleotides include DNA.

98. The lipid nanoparticles according to claim 95, wherein the one or more polynucleotides include DNA and RNA.

99. A pharmaceutical composition comprising lipid nanoparticles according to any one of claims 79 to 98 and a pharmaceutically acceptable excipient.

100. A pharmaceutical composition comprising lipid nanoparticles according to any one of claims 95 to 98 and a pharmaceutically acceptable excipient.

101. A method for delivering polynucleotides to target cells or tissues, comprising administering an effective amount of lipid nanoparticles according to any one of claims 79 to 98 or the pharmaceutical composition according to claim 100 to the target.

102. The method according to claim 101, wherein the cells or tissues include extrahepatic cells or tissues.

103. The method according to claim 101, wherein the cells or tissue include brain cells or tissue.

104. The method according to claim 101, wherein the cells or tissue include lung cells or tissue.

105. The method according to claim 101, wherein the cells or tissue include bone marrow cells or tissue.

106. The method according to claim 101, wherein the cells or tissue include spleen cells or tissue.

107. The method according to claim 101, wherein the cells or tissues include muscle cells or tissues.

108. The method according to claim 101, wherein the cells or tissue include kidney cells or tissue.

109. The method according to claim 101, wherein the cells or tissues include cardiac cells or tissues.

110. The method according to claim 101, wherein the cells or tissue include pancreatic cells or tissue.

111. The method according to claim 101, wherein the cells or tissues include immune cells or tissues.

112. A method for treating a target disease, comprising administering a therapeutically effective amount of the pharmaceutical composition according to claim 99 or 100 to the target.

113. A method for producing a therapeutic composition, comprising encapsulating an active agent within lipid nanoparticles, wherein the lipid nanoparticles contain a compound according to any one of claims 1 to 78.

114. A method for producing a vaccine or prophylactic composition, comprising encapsulating an active agent within lipid nanoparticles, wherein the lipid nanoparticles contain a compound according to any one of claims 1 to 78.

115. The method according to claim 113 or 114, wherein the active agent comprises DNA.

116. The method according to claim 113 or 114, wherein the active agent comprises RNA.

117. The method according to claim 113 or 114, wherein the active agent comprises DNA and RNA.

118. Lipid nanoparticles according to any one of claims 79 to 98 or the pharmaceutical composition according to claim 99 or 100 for use in the delivery of polynucleotides to target extrahepatic cells or tissues.

119. Use of lipid nanoparticles according to any one of claims 79 to 98 or the pharmaceutical composition according to claim 99 or 100 in the manufacture of a pharmaceutical for delivering polynucleotides to target extrahepatic cells or tissues.

120. Lipid nanoparticles according to any one of claims 79 to 98 or a pharmaceutical composition according to claim 99 or 100 for use in the treatment of a target disease.

121. Use of lipid nanoparticles according to any one of claims 79 to 98 or the pharmaceutical composition according to claim 99 or 100 in the manufacture of a pharmaceutical for the treatment of a target disease.

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