Novel lipid compound and lipid nanoparticle composition containing the same

Abstract

The present invention relates to a novel lipid compound and a lipid nanoparticle composition containing the same. More specifically, the lipid nanoparticle composition contains an ionizable lipid, a helper lipid, a PEG-lipid, and an additive, and contains a biocompatible vitamin-based novel lipid compound and a helper lipid containing a neutral lipid, which can mitigate changes in the delivery mechanism and side effects, and can improve protein expression efficiency.

Description

[Technical Field]

[0001] The present invention relates to novel lipid compounds and lipid nanoparticle compositions containing the same. [Background technology]

[0002] Nucleic acid-based medicines, which began over 40 years ago as a method of injecting plasmid DNA into the human body to stimulate the production of missing proteins, have since been developed into a variety of drugs, including antigens, decoys, antisense, siRNA, and miRNA, which inhibit gene transcription and translation. Nucleic acid-based medicines target DNA or RNA rather than proteins and have attracted attention as personalized therapeutics by binding complementary to specific DNA or RNA sequences. Nucleic acid-based medicines are used not only as therapeutic preparations but also as preventative preparations that inject genes capable of expressing antigens against specific diseases. Gene-based vaccines are divided into DNA vaccines, RNA vaccines, and viral vector vaccines. Of these, RNA vaccines eliminate the risk of infection associated with viral vector-based vaccines and the potential risks of genetic mutation associated with DNA vaccines. They also offer the advantage of rapid development and have garnered attention as an effective response to the COVID-19 outbreak that emerged in 2019.

[0003] However, nucleic acid-based drugs are easily degraded by nucleases in the human body and are negatively charged macromolecules that cannot be easily delivered into cells. Therefore, a stable and efficient method for delivering them to the desired location is needed. Nucleic acid delivery systems based on a variety of materials, including lipids, polymers, dendrimers, and inorganic metal materials, have been reported. However, lipid nanoparticles were used in patisiran, the first siRNA new drug approved by the FDA in 2018, as well as the mRNA vaccine for COVID-19 approved for emergency use in 2020. Currently, lipid nanoparticles are generally formed by mixing four components in a set ratio: ionized lipid, phospholipid (helper lipid), cholesterol (structure-maintaining lipid), and PEG-lipid.

[0004] As new siRNA drugs and mRNA vaccines have been commercialized, lipid nanoparticle-based delivery vehicles have faced various challenges in the delivery field of traditional nucleic acid drugs, such as improving stability and cell permeability, as well as various other issues that must be resolved, such as ensuring targeting, ease of storage and distribution, mitigating side effects, reducing costs, and addressing breakthrough infections. Therefore, the nucleic acid drug market is in need of the development of lipid nanoparticles that can solve these new challenges. Summary of the Invention [Problem to be solved by the invention]

[0005] The object of the present invention is to provide novel vitamin-based ionizable lipids and helper lipid compounds, and lipid nanoparticle compositions containing the same that are biocompatible and can improve the efficiency and sustainability of protein expression. [Means for solving the problem]

[0006] To achieve the above object, the present invention provides a compound selected from the group consisting of a compound represented by the following chemical formula 1, a stereoisomer thereof, a racemate thereof, and a pharmaceutically acceptable salt thereof:

[0007] [ka] In the above Chemical Formula 1, R1 and R2 are the same or different and each represent hydrogen or a saturated hydrocarbon having 1 to 2 carbon atoms; X1 is O, NH, or S; m1, m2, and m3 are the same or different and each represent an integer of 1 to 3; and A is hydrogen or a compound represented by the following Chemical Formula 1-1:

[0008] [ka]

[0009] In the above Chemical Formula 1-1, Z is NR 1 R 2 a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, provided that R 1 and R 2 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocycle or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, the hydrocarbon may or may not contain an ester, ether, amide, carbamate, carbonate or disulfide bond, X2 is O or S, Y is CH2, NH or O, and n1 is an integer of 0 to 3;

[0010] B is a compound represented by the following chemical formula 1-2, or NR 1 'R 2 ', but the R 1 ' and R 2 ' are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R 1 ' and R 2 ' are linked to each other to form a heterocyclic or aromatic ring having 3 to 8 carbon atoms,

[0011] [ka]

[0012] In the formula 1-2, R3 and R4 are the same or different and are linear or branched saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, wherein the hydrocarbons may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond, and E is hydrogen or a compound represented by the following formula 1-3:

[0013] [ka]

[0014] In Formulas 1-3, X3 is O or S, n2 is an integer of 0 to 3, and R5 is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond and may be substituted or unsubstituted with thiolane or dithiolane.

[0015] The present invention provides a lipid nanoparticle composition comprising the compound.

[0016] The present invention provides a lipid nanoparticle composition comprising one or more compounds selected from the compound represented by Chemical Formula 1, a stereoisomer thereof, a racemate thereof, or a pharmaceutically acceptable salt thereof.

[0017] The present invention provides a composition for drug delivery comprising the lipid nanoparticle composition described above; and a therapeutic or prophylactic formulation.

[0018] The present invention also provides an immune enhancing composition comprising the lipid nanoparticle composition. [Effects of the Invention]

[0019] According to the present invention, lipid nanoparticles that stably express proteins can be constructed by incorporating a novel biocompatible vitamin-based lipid compound, and it is expected that the toxicity problem of lipid nanoparticles, which is often caused by ionized lipids, can also be resolved. [Brief explanation of the drawings]

[0020] [Figure 1] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 127, 130) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98, 126) into rat ears (id). [Figure 2] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 131, 132, 133) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 128) into rat ears (id). [Figure 3] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 136, 138, 139) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98, 128) into rat ears (id). [Figure 4] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of novel cholesterol-substitute lipid nanoparticles (LNP 109, 111, 114) containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98) into rat ears (id). [Figure 5] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of novel cholesterol-substituting lipid nanoparticles (LNP 110, 115, 117, 118) containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98) into rat ears (id). [Figure 6] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of novel cholesterol-substitute lipid nanoparticles (LNP 119, 123) containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98) into rat ears (id). [Figure 7] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of a novel cholesterol-substitute lipid nanoparticle (LNP 168) containing Renilla luciferase (R / L) mRNA and a control lipid nanoparticle (LNP 98) into the ears of rats (id). [Figure 8] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of a novel cholesterol-substitute lipid nanoparticle (LNP 200) containing Renilla luciferase (R / L) mRNA and a control lipid nanoparticle (LNP 126) into rat ears (id). [Figure 9] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 146, 147) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98) into rat ears (id). [Figure 10] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 152, 153) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98, 128) into rat ears (id). [Figure 11] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 159, 160) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98, 128) into rat ears (id). [Figure 12]The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 166, 167) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 98, 128) into rat ears (id). [Figure 13] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 178, 179, 181, 182) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 138, 139) into rat ears (id). [Figure 14] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 185, 187) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126) into rat ears (id). [Figure 15] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 192, 193) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 16] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 205, 208, 209, 210) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 187) into rat ears (id). [Figure 17] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 211, 212, 213, 214, 216, 217) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 166) into rat ears (id). [Figure 18]The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 224, 225, 226) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126) into rat ears (id). [Figure 19] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 227, 228, 229, 230, 231, 232) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126) into rat ears (id). [Figure 20] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 256, 257) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 21] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 278, 279, 280, 281) with a novel ionized lipid composition containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 22] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 291, 292, 293, 294, 295, 296) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 23]The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of novel cholesterol-substituting lipid nanoparticles (LNP 310, 311, 312, 313, 314, 315) containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 24] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 323, 324, 325, 326, 327, 328) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 25] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 329, 330, 331, 332, 333, 334) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 26] The amount of Renilla luciferase (R / L) expressed was analyzed 6 and 24 hours after injection of lipid nanoparticles (LNP 335, 336, 337, 338) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128) into rat ears (id). [Figure 27] The amount of Renilla luciferase (R / L) expressed was analyzed 6 hours after injection into rat ears (id) with lipid nanoparticles (LNP 412, 417, 422, 425) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA and control lipid nanoparticles (LNP 126, 128, 295, 334). [Figure 28]After formulating mRNA encoding the influenza virus surface antigen (LNP 126, 181, 187, 200), rats were immunized intramuscularly, and the HA-specific antibody response in the blood was confirmed through IgG1 (a, c) and IgG2a (b, d) levels two weeks after the primary immunization (a-b) and secondary immunization (c-d). [Figure 29] After formulating mRNA encoding the influenza virus surface antigen (LNP 126, 128, 230, 231, 232), rats were immunized intramuscularly, and the HA-specific antibody response in the blood was confirmed through IgG1 (a, c) and IgG2a (b, d) levels two weeks after the primary immunization (a-b) and secondary immunization (c-d). [Figure 30] After formulating mRNA encoding the influenza virus surface antigen (LNP 128, 295), rats were immunized intramuscularly, and the HA-specific antibody response in the blood was confirmed through IgG1 (a, c) and IgG2a (b, d) levels two weeks after the primary immunization (a-b) and secondary immunization (c-d). [Figure 31] After formulating mRNA encoding Renilla luciferase (R / L) (LNP 126, 159, 211, 224, 227, 230, 291, 294, 310, 313, 323, 325, 327, 329, 331, 333, 335, 337), it was administered ID to the ear of ICR rats, and MCP-1 concentrations were measured in blood collected 6 hours later. [Figure 32] After formulating mRNA encoding Renilla luciferase (R / L) (LNP 128, 160, 212, 225, 228, 231, 292, 295, 311, 314, 324, 326, 328, 330, 332, 334, 336, 338), it was administered ID to the ear of ICR rats, and MCP-1 concentrations were measured in blood collected 6 hours later. DETAILED DESCRIPTION OF THE INVENTION

[0021] The present invention will now be described in more detail.

[0022] The present invention provides a compound selected from the group consisting of a compound represented by the following chemical formula 1, a stereoisomer thereof, a racemate thereof, and a pharmaceutically acceptable salt thereof:

[0023] [ka] In the above Chemical Formula 1, R1 and R2 are the same or different and each represent hydrogen or a saturated hydrocarbon having 1 to 2 carbon atoms; X1 is O, NH, or S; m1, m2, and m3 are the same or different and each represent an integer of 1 to 3; and A is hydrogen or a compound represented by the following Chemical Formula 1-1:

[0024] [ka]

[0025] In the above Chemical Formula 1-1, Z is NR 1 R 2 a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, provided that R 1 and R 2 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocycle or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, the hydrocarbon may or may not contain an ester, ether, amide, carbamate, carbonate or disulfide bond, X2 is O or S, Y is CH2, NH or O, and n1 is an integer of 0 to 3;

[0026] B is a compound represented by the following chemical formula 1-2, or NR 1 'R 2 ', but the R 1 ' and R 2' are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R 1 ' and R 2 ' are linked to each other to form a heterocyclic or aromatic ring having 3 to 8 carbon atoms,

[0027] [ka]

[0028] In the formula 1-2, R3 and R4 are the same or different and are linear or branched saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, wherein the hydrocarbons may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond, and E is hydrogen or a compound represented by the following formula 1-3:

[0029] [ka]

[0030] In Formulas 1-3, X3 is O or S, n2 is an integer of 0 to 3, and R5 is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond and may be substituted or unsubstituted with thiolane or dithiolane.

[0031] The heterocyclic compound may be selected from the group consisting of thiophene, furan, pyrazole, pyridine, pyran, oxazine, thiazine, morpholine, pyrrolidine, piperidine, piperazine, pyrazole, pyridine, and dithiolane, each of which may be substituted or unsubstituted with an alkyl group having 1 to 4 carbon atoms.

[0032] The aromatic ring compound can also be benzene substituted or unsubstituted with a di(C1-C4) alkylamino(C1-2) alkyl group.

[0033] Specifically, the compound represented by Chemical Formula 1 is also a compound represented by Chemical Formula 2 below:

[0034] [ka]

[0035] In the above chemical formula 2, A1 is hydrogen or a compound represented by the following chemical formula 2-1:

[0036] [ka]

[0037] In the above Chemical Formula 2-1, Z1 is NR 3 R 4 or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, 3 and R 4 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocycle or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and n3 is an integer of 0 to 3;

[0038] E1 is hydrogen or a compound represented by the following chemical formula 2-2,

[0039] [ka]

[0040] In Formula 2-2, R8 is a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, wherein the hydrocarbon may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond, and may or may not be substituted with dithiolane;

[0041] R6 and R7 may be the same or different and may be linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms.

[0042] The compound represented by Chemical Formula 1 is also a compound represented by Chemical Formula 3 below:

[0043] [ka] In the above Chemical Formula 3, A2 is hydrogen or a compound represented by the following Chemical Formula 3-1:

[0044] [ka]

[0045] In the above Chemical Formula 3-1, Z2 is NR 5 R 6 or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, 5 and R 6 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocycle or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and n4 is an integer of 0 to 3;

[0046] R9 and R 10 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, wherein the hydrocarbons may or may not contain ester, ether, amide, carbamate, carbonate, or disulfide bonds, and o1 and o2 are the same or different and are integers of 2 to 10.

[0047] The compound represented by Chemical Formula 1 is also a compound represented by Chemical Formula 4:

[0048] [ka] In the above Chemical Formula 4, X1' is O, NH or S, m4 and m5 are the same or different and each represent an integer of 0 to 3, and A3 is a compound represented by the following Chemical Formula 4-1:

[0049] [ka]

[0050] In the above Chemical Formula 4-1, Z3 is NR 7 R 8 a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, provided that R 7 and R 8 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocycle or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and the hydrocarbon may or may not contain an ester, ether, amide, carbamate, carbonate or disulfide bond; n5 is an integer of 0 to 3;

[0051] B1 is a compound represented by the following chemical formula 4-2: NR 9 R 10 or a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, 9 and R 10 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R 9 and R 10are linked to each other to form a heterocycle or aromatic ring having 3 to 8 carbon atoms,

[0052] [ka]

[0053] In the above Chemical Formula 4-2, R 12 and R 13 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds;

[0054] R 11 is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds, and is substituted or unsubstituted with dithiolane.

[0055] More specifically, the compound represented by Chemical Formula 2 is also a compound represented by Chemical Formula 5 or Chemical Formula 6:

[0056] [ka] In the above formula 5, R 14 and R 15 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, which may or may not contain ester, ether, amide, carbamate, carbonate, or disulfide bonds.

[0057] [ka] In the above Chemical Formula 6, Z4 is NR 11 R 12 or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, 11 and R12 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocyclic or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl;

[0058] R 16 and R 17 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, which may or may not contain ester, ether, amide, carbamate, carbonate, or disulfide bonds.

[0059] The compound represented by Chemical Formula 3 is also a compound represented by Chemical Formula 7:

[0060] [ka] In the above formula 7, R 18 and R 19 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, wherein the hydrocarbons may or may not contain ester, ether, amide, carbamate, carbonate, or disulfide bonds, and o1' and o2' are the same or different and are integers of 3 to 8.

[0061] The compound represented by Chemical Formula 4 is also a compound represented by Chemical Formula 8 or Chemical Formula 9:

[0062] [ka] In the above Chemical Formula 8, m6 is an integer of 0 to 3, and R 20 and R 21 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; 20 and R 21 are linked to each other to form a heterocycle having 3 to 8 carbon atoms,

[0063] R 22 and R 23 are the same or different and are each hydrogen, a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without an ester, ether, amide, carbamate, carbonate, or disulfide bond; R 24 is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester or disulfide bond, and which may or may not be substituted with dithiolane.

[0064] [ka] In the above Chemical Formula 9, m7 and m8 are the same or different and each represent an integer of 0 to 3; R 25 is a linear or branched saturated or unsaturated hydrocarbon having 1 to 6 carbon atoms, wherein the hydrocarbon may or may not contain an ester bond or a disulfide bond; R 26 and R 27 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms; R 26 and R 27 are linked together to form a heterocycle or aromatic ring having 3 to 8 carbon atoms, and R 28 is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester or disulfide bond, and which may or may not be substituted with dithiolane.

[0065] For example, the compound represented by Chemical Formula 5 is also a helper lipid represented by Chemical Formula 10 or Chemical Formula 11 below:

[0066] [ka]

[0067] The compound represented by Chemical Formula 6 may also be an ionized lipid represented by any one selected from the group consisting of Chemical Formulas 12 to 23 below: [ka] [ka]

[0068] The compound represented by Chemical Formula 7 is also an ionized lipid represented by Chemical Formula 24: [ka]

[0069] The compound represented by Chemical Formula 8 may also be an ionized lipid represented by any one selected from the group consisting of Chemical Formulas 25 to 28 and Chemical Formulas 37 to 38: [ka]

[0070] The compound represented by Chemical Formula 9 may also be an ionized lipid represented by any one selected from the group consisting of Chemical Formulas 29 to 36 and Chemical Formulas 39 to 40: [ka] [ka]

[0071] The present invention also provides a lipid nanoparticle composition comprising a compound selected from the group consisting of a compound represented by the above-mentioned Chemical Formula 1, a stereoisomer thereof, a racemate thereof, or a pharmaceutically acceptable salt thereof.

[0072] The compound may also be an ionizable lipid or a helper lipid.

[0073] The corresponding features can be substituted in the above-mentioned parts.

[0074] The lipid nanoparticle composition comprises one or more selected from the group consisting of a helper lipid, a structure-retaining lipid, a PEG-lipid, and an additive.

[0075] The helper lipids include 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundeca Hemisuccinoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearoyl-phosphatidyl- and one or more selected from the group consisting of dimethicone, dimethicone diol, dimethicone diol (DSPE), dipalmitoyl-sn-glycero-3-phosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, and lysophosphatidylethanolamine (LPE).

[0076] The structure-retaining lipid may be one or more selected from the group consisting of cholesterol, bile acid derivatives including butyl lithocholate, cholanic acid derivatives, lithocholic acid derivatives, flavonoids, vitamin A and its derivatives, vitamin E, vitamin K, coenzyme Q10, and β-carotene.

[0077] The lithocholic acid derivative is also a compound represented by the following chemical formula 41:

[0078] [ka]

[0079] In the above formula 41, R 29 and R 30 are the same or different and each is a linear or branched, saturated or unsaturated hydrocarbon having 1 to 22 carbon atoms, 29 and R 30 At least one of the above is a saturated or unsaturated hydrocarbon having 2 to 20 carbon atoms.

[0080] Specifically, the compound represented by the formula 41 is R 29 is hydrogen and R 30 is also a compound represented by formula 42 having an alkyl group having four carbon atoms:

[0081] [ka]

[0082] The PEG-lipid may be one or more selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

[0083] The additive may also be a trehalose derivative of formula 43:

[0084] [ka]

[0085] In the above formula 43, R 31 and R 32 are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms.

[0086] Specifically, the compound represented by Chemical Formula 43 is also a compound represented by Chemical Formula 44:

[0087] [ka]

[0088] Specifically, the lipid nanoparticle composition contains an ionizable lipid, a helper lipid, a structure-retaining lipid, a PEG-lipid, and an additive, and may contain the ionizable lipid in a ratio of 20 to 60 mol %, 5 to 40 mol % of the helper lipid, 25 to 45 mol % of the structure-retaining lipid, 1 to 3 mol % of the PEG-lipid, and 0 to 25 mol % of the additive.

[0089] More specifically, the composition may contain 20 to 50 mol % of ionizable lipid, 5 to 15 mol % of helper lipid, 25 to 45 mol % of structure-retaining lipid, 1 to 3 mol % of PEG-lipid, and 0 to 25 mol % of additive.

[0090] More specifically, the composition may include a compound of any one of Formulas 12 to 40 as the ionizable lipid; DSPC, DOPE, or a compound of Formula 10 or 11 as the helper lipid; cholesterol, a lithocholic acid derivative, or a cholanic acid derivative as the structure-retaining lipid; myristoyl diglyceride (DMG)-PEG as the PEG-lipid; and 6,6'-trehalose dioleate as the additive.

[0091] The lipid nanoparticle compositions of the present invention may further comprise therapeutic and / or prophylactic agents.

[0092] Said therapeutic and / or prophylactic preparations may also be vaccines or compounds capable of inducing an immune response.

[0093] The therapeutic and / or prophylactic formulation may be selected from the group consisting of DNA, interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof.

[0094] The encapsulation efficiency of the therapeutic and / or prophylactic formulation is also independently at least 50% or greater.

[0095] The wt / wt ratio of lipid component to said therapeutic and / or prophylactic formulation may also be from about 10:1 to about 60:1.

[0096] The therapeutic and / or prophylactic formulations also have an N:P ratio of about 2:1 to about 30:1, which is the number of ionizable nitrogens in the ionizable lipid divided by the number of phosphate groups in the nucleic acid molecule.

[0097] The present invention also provides a composition for drug delivery comprising the lipid nanoparticle composition and a therapeutic and / or prophylactic formulation described above.

[0098] The corresponding features can be substituted in the above-mentioned parts.

[0099] The present invention also provides a method for delivering therapeutic and / or prophylactic agents to mammalian cells via the drug delivery composition.

[0100] The corresponding features can be substituted in the above-mentioned parts.

[0101] A method for delivering the therapeutic and / or prophylactic formulation to mammalian cells includes administering the lipid nanoparticle composition to a subject, wherein the administration brings the cells into contact with the nanoparticle composition, and the therapeutic and / or prophylactic formulation can be delivered to the cells.

[0102] The mammalian cell is from a mammal.

[0103] The mammal is a human.

[0104] The drug delivery composition may be administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. A dose of about 0.01 to about 10 mg / kg of the therapeutic and / or prophylactic formulation is administered to the mammal.

[0105] The present invention also provides a method for producing a polypeptide of interest in mammalian cells.

[0106] The method for producing a polypeptide of interest in the mammalian cell includes contacting the cell with a drug delivery composition comprising a lipid nanoparticle composition and a therapeutic and / or prophylactic agent, wherein the therapeutic and / or prophylactic agent is mRNA, the mRNA encoding the polypeptide of interest, such that the mRNA is translated in the cell to produce the polypeptide of interest.

[0107] The corresponding features can be substituted in the above-mentioned parts.

[0108] The present invention also provides an immune enhancing composition comprising the lipid nanoparticle composition described above.

[0109] The corresponding features can be substituted in the above-mentioned parts.

[0110] The present invention will be described in detail below with reference to examples to aid in understanding the present invention. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

[0111] [Example 1] Synthesis of pantothenic acid-based helper lipid

[0112] [ka]

[0113] 1-1-1,4-(((4-Methoxybenzyl)oxy)methyl)-2,2-dimethyl-1,3-dioxolane (47)

[0114] Sodium hydride (0.50 g, 12 mmol, 60% dispersion in mineral oil) was stirred in anhydrous DMF (10 mL) and cooled to 0°C. Solketal (compound 45) (1 g, 7.55 mmol) was then added dropwise to the solution, which was then warmed to room temperature and stirred for 10 minutes under argon gas. The reaction mixture was then cooled to 0°C again, and p-methoxybenzyl chloride (compound 46) (1.75, 11 mmol) was added dropwise. The mixture was then vigorously stirred at room temperature for 12 hours. Upon completion of the reaction, the reaction mixture was cooled to 0°C, anhydrous methanol (1 mL) was slowly added, and the mixture was diluted with EtOAc (30 mL). The organic layer was washed sequentially with water (2 × 20 mL) and saturated aqueous NaCl (20 mL). The organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was concentrated using a rotary evaporator. It was then purified by flash column chromatography (SiO2, EtOAc / hexane 1:9->2:8) to give compound 47 (6.34 g, 93%) as a clear oil.

[0115] 1 H NMR (CDCl3, 400MHz): δ1.36(s, 3H), 1.43(s, 3H), 3.50-3.61(m, 1H), 3.60-3.72(m, 1H), 3.75-3.82(m, 1H) , 3.83(s, 3H), 3.91-4.05(m, 1H), 4.16-4.26(m, 1H), 4.56(s, 3H), 6.83(d, J=7Hz, 2H), 7.24(d, J=7Hz, 2H); 13 C NMR (CDCl3, 100MHz): δ25.4, 26.76, 55.2, 66.8, 70.7, 73.2, 74.7, 109.4, 113.8, 114.2, 130.2, 130.2, 129.4.

[0116] 1-1-2,3-((4-Methoxybenzyl)oxy)propane-1,2-diol [3-((4-Methoxybenzyl)oxy)propane-1,2-diol] (48)

[0117] The synthesized acetal compound 47 (2 g, 8 mmol) was dissolved in 80% aqueous AcOH (40 mL) and heated to 65 °C with vigorously stirring for 1.5 h. After confirming the completion of the reaction by TLC (SiO2, hexane / EtOAc 1:9, CAM stain), the reaction mixture was removed using a rotary evaporator with toluene (30 mL x 2) and purified by column chromatography (SiO2, EtOAc / hexane 8:2) to give compound 48 (6.34 g, 84%) as a clear oil.

[0118] 1 H NMR (CDCl3, 400MHz): δ7.24(d, J=8.8Hz), 6.88(d, J=8.8Hz), 4.47(s, 2H), 3.84-3.89(m, 1H), 3.7 8(s, 3H), 3.67(dd, J=11.5, 3.9Hz), 3.59(dd, J=11.6, 5.7Hz, 1H), 3.47-3.53(m, 2H), 2.69(s, 2H); 13 C NMR (CDCl3, 100MHz): δ159.4, 129.8, 129.5, 113.9, 73.1, 71.3, 70.5, 64.1, 55.3.

[0119] 1-1-3,3-((4-Methoxybenzyl)oxy)propane-1,2-diyl dioleate (49)

[0120] Oleic acid (665 mg, 2.5 equiv.) was placed in a reaction vessel and dissolved in DCM (40 mL). EDCI·HCl (580 mg, 2.5 equiv.) and 4-dimethylaminopyridine (DMAP; 55 mg, 0.3 equiv.) were added and the mixture was stirred vigorously under argon gas at 5°C for 10 minutes. Next, a solution of 3-((4-methoxybenzyl)oxy)propane-1,2-diol (200 mg, 1 equiv.) dissolved in DCM (10 mL) was added dropwise while maintaining the temperature at 5°C, and the mixture was stirred at room temperature for 24 hours. Once the reaction was confirmed to be complete by TLC (SiO2, EtOAc / hexane 4:6), DCM (50 mL) was added to the reaction mixture, and the organic layer was washed with saturated aqueous NaHCO3 (50 mL x 2) and saturated aqueous NaCl (20 mL). After drying over anhydrous MgSO4, the filtered filtrate was concentrated using a rotary evaporator. Compound 49 (594 mg, 85%) was then purified by column chromatography (SiO2, EtOAc / hexane 1:9) to give a clear solution.

[0121] 1 H NMR (CDCl3, 400MHz): δ0.86-0.89(t, J=5.6Hz, 6H), 1.26-1.29(m, 41H), 1. 59-1.60(m, 5H), 1.98-2.00(m, 8H), 2.01-2.33(m, 4H), 3.54-3.55(m, 2H), 3.56-3.80(s, 3H), 4.14-4.34(m, 2H), 4.47(dd, J=3.6, 4.8Hz, 2H), 5.21-5 .22(m, 2H), 5.32-5.35(m, 4H), 7.21(d, J=8.8Hz, 2H), 7.26(d, J=9Hz, 2H); 13C NMR (CDCl3, 100MHz): δ14.11, 22.69, 24.88, 24.96, 27.19, 27.24, 29.08, 29.11, 29.14, 29.21, 29.33, 29.54, 29.73, 29.78, 31.92, 34.11, 34.33, 55.26, 62.71, 67.93, 70.06, 72.98, 113.83, 129.30, 129.72, 129.80, 130.02, 159.34, 173.08, 173.38.

[0122] 1-1-4,3-Hydroxypropane-1,2-diyl dioleate [3-Hydroxypropane-1,2-diyl dioleate(1,2-diolein;1,2-dioleoyl-rac-glycerol)] (50)

[0123] Compound 49 (210 mg, 0.28 mmol, 1 equiv.) was added to a DCM / distilled water mixture (20:1, 20 mL) and stirred for 5 minutes. DDQ (96.48 mg, 0.368 mmol, 1.5 equiv.) was then added and stirred at room temperature for 2 hours. After the reaction was complete, the solid was filtered off, and NaHCO3 (200 mg) was added to the filtrate. The layers were separated. The aqueous layer was extracted with DCM (25 mL x 2). The combined organic layer was washed with saturated aqueous NaHCO3 (25 mL x 2) and saturated aqueous NaCl (25 mL) and then dried over anhydrous MgSO4. The filtered organic layer was then purified by flash column chromatography (SiO2, 1:9 EtOAc / hexane) to obtain compound 50 (125 mg, 71%) as a clear oil.

[0124] 1 H NMR (CDCl3, 400MHz): δ0.83-0.89(m, 6H), 1.25-1.37(m, 40H), 1.41-1.44(m, 4H), 1.99-2.07(m, 8H), 2.30-2 .36(m, 4H), 3.72-3.73(m, 2H), 4.21-4.25(m, 1H), 4.30-4.33(m, 2H), 5.07-5.09(m, 1H), 5.32-5.39(m, 4H); 13C NMR (CDCl3, 100MHz): δ27.31, 27.42, 29.21, 29.32, 29.43, 29.52, 29.61, 29.62, 29.84, 29.85, 29.87, 30.31, 31.56, 31 .71, 32.04, 32.05, 33.85, 33.95, 34.21, 34.42, 34.64, 61.71, 62.27, 72.35, 129.87, 130.14, 130.34, 173.51, 173.81.

[0125] [ka]

[0126] 1-2-1.(R)-3-(2,2,5,5-Tetramethyl-1,3-dioxane-4-carboxamido)propanoic acid (52)

[0127] D-pantothenic acid hemicalcium salt (compound 51, 5 g, 1 equiv.) and 2,2-dimethoxypropane (DMP; 60 mL) were stirred in a reaction vessel, while p-toluenesulfonic acid (PTSA, 3.97 g, 1.1 equiv.) was added in one portion. The reaction mixture was stirred at room temperature for 20 hours, and the resulting white solid was washed with acetone and filtered. The filtrate was distilled under reduced pressure, and the resulting yellow solid was washed with warm n-hexane to obtain compound 52 (5.45 g, 100%) as a white solid.

[0128] 1 H NMR (CDCl3, 400MHz): δ0.98(s, 3H), 1.04(s, 3H), 1.43(s, 3H), 1.46(s, 3H), 2.62(dt, J=6.0, 2.0, 2H), 3.29(d, J=1.5, 1H), 3.45-3.53(m, 1H), 3.56-3.64(m, 1H), 3.70(d, J=11.5, 1H), 4.11(s, 1H), 7.05(app bs, 1H); 13C NMR (CDCl3, 100MHz): δ18.7, 18.8, 22.0, 29.4, 30.9, 33.0, 33.9, 34.1, 71.4, 77.1, 99.1, 170.2.

[0129] 1-2-2,3-((3-(2,2,5,5-Tetramethyl-1,3-dioxane-4-carboxamido)propanoyl)oxy)propane-1,2-diyl dioleate] (53)

[0130] Carboxylic acid compound 52 (62 mg, 1.5 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (46 mg, 1.5 equiv.) and DMAP (4 mg, 0.2 equiv.) were added, and the reaction mixture was vigorously stirred under argon gas at 5°C for 10 min. Compound 50 (100 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5°C and stirred at room temperature for 12 h. After confirming completion of the reaction using TLC (SiO2, EtOAc / hexane 2:8), an additional 50 mL of DCM was added, and the organic layer was washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The organic layer was dried over anhydrous MgSO4 to remove water, and the filtered filtrate was distilled under reduced pressure to remove the solvent. The remaining mixture was separated by flash column chromatography (SiO2, EtOAc / hexane 3:7) to obtain compound 53 (100 mg, 72%) as a clear liquid.

[0131] 1 H NMR (CDCl3, 400MHz): δ0.87(t, J=7.1Hz, 6H), 0.95(s, 3H), 1.03(s, 3H), 1.25-1.30(m, 43H), 1.41(s, 3H), 1.45(s, 3H), 1.61(brs, 4H), 1.98-2.01(m, 8H), 2.28-2.32(m, 4H), 2.56-2.58(t , J=6.5Hz, 2H), 3.28(d, J=11.5Hz, 1H), 3.48-3.58(m, 2H), 3.66(d, J=11.5Hz, 1H), 4.07(s, 1 H), 4.11-4.17(m, 2H), 4.26-4.34(m, 2H), 5.24-5.31(m, 1H), 5.32-5.35(m, 4H), 7.25(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.02, 18.58, 18.59, 18.73, 21.99, 22.58, 24.73, 24.76, 2 7.73, 24.76, 29.22, 29.36, 29.42, 29.61, 29.66, 31.79, 31.80, 32.86, 33.78, 33.8 0, 33.91, 34.03, 34.05, 34.07, 61.88, 68.64, 71.37, 76.71, 76.93, 77.14, 98.92, 129.58, 126.60, 129.92, 129.93, 136.71, 171.55, 171.57, 172.76, 172.78, 173.13.

[0132] 1-2-3,3-((3-(2,4-dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl dioleate (Compound 10)

[0133] Compound 53 (100 mg) was placed in a reaction vessel and dissolved in a mixture of AcOH / H2O (2:1, 20 mL) and stirred at room temperature for 40 minutes. Saturated aqueous NaHCO3 (50 mL) was added to the reaction mixture, which was then extracted with EtOAc (50 mL x 1, 20 mL x 2). The combined EtOAc layers were washed with saturated aqueous NaCl (20 mL), then removed with anhydrous MgSO4. The filtrate was filtered and the solvent was removed by distillation under reduced pressure. The remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:1) to give compound 10 (36.22 mg, 38%) as a clear oil.

[0134] 1 H NMR (CDCl3, 400MHz): δ0.87(t, J=7.1Hz, 6H), 0.90(d, J=12Hz, 3H), 1.03(d, J=12H z, 3H), 1.23-1.34(m, 42H), 1.60(s, 4H), 1.98-2.01(m, 8H), 2.30-2.33(m, 4H), 2. 55-2.56(m, 2H), 3.15-3.21(m, 1H), 3.47-3.65(m, 3H), 3.75-3.78(m, 1H), 4.00-4 .06(m, 1H), 4.16-4.17(m, 1H), 4.20-4.39(m, 4H), 5.30-5.36(m, 5H), 7.22(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.01, 19.86, 20.11, 21.61, 21.73, 22.58, 24.71, 24.72, 24.77, 27 .07, 27.12, 28.92, 28.94, 28.98, 29.01, 29.02, 29.07, 23.13, 29.21, 29.23, 29.42, 29.60, 29.66, 31.80, 33.89, 33.93, 33.94, 34.09, 34.41, 39.27, 61.90, 61.95, 62.51, 62.60, 68.71, 68.75, 71.05, 71.09, 129.57, 129.58, 129.94, 129.95, 171.76, 173.09, 173.38, 173.56.

[0135] [ka]

[0136] 1-3-1,2,3-Dihydroxypropyl acetate (55)

[0137] Propane-1,2,3-triol (compound 54) (300 mg, 1 equiv.) and ethanoic anhydride (432 mg, 1.3 equiv.) were dissolved in anhydrous acetonitrile (20 mL). Tetra-n-butylammonium acetate (TBAAc; 687 mg, 0.7 equiv.) was added, and the reaction mixture was stirred at 50 °C for 7 h. After the reaction was completed, the reaction mixture was loaded by solid deposition and purified by flash column chromatography (SiO2, n-hexane / EtOAc 5:5) to give compound 55 (297 mg, 68%) as a colorless oil.

[0138] 1H NMR (CDCl3, 400MHz): δ2.11(s, 3H), 2.7-2.8(bs, 1H), 3.09-3.19(bs, 1H), 3.59(dd, 1H, J= 6, 11.6Hz), 3.69(dd, 1H, J=3.6, 11.6Hz), 3.93(pentet, 1H, J=4.8Hz), 4.22-4.12(m, 2H); 13 C NMR (CDCl3, 100MHz): δ20.83, 63.31, 65.29, 70.12, 171.56.

[0139] 1-3-2,3-Acetoxypropane-1,2-diyl distearate (56)

[0140] Stearic acid (1060 mg, 2.5 equiv.) was dissolved in DCM (40 mL) in a reaction vessel, and EDCI·HCl (580 mg, 2.5 equiv.) and DMAP (55 mg, 0.3 equiv.) were added. The reaction mixture was vigorously stirred under argon gas at 5°C for 10 minutes, followed by dropwise addition of compound 55 (200 mg, 1 equiv.) in DCM (20 mL) at 5°C and stirring at room temperature for 22 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 4:6), DCM (50 mL) was added to the reaction mixture, which was then placed in a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The organic layer was combined and dehydrated using anhydrous MgSO4. The filtrate was then evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 2:8) to give compound 56 (846 mg, 85%) as a clear liquid.

[0141] 1 H NMR (CDCl3, 400MHz): δ0.87(t, J=6Hz, 6H), 1.01(m, 9H), 1.21(m, 56H), 1.62(m, 4H), 2.08(s, 1H), 2.25(m, 4H), 3.74(m, 2H), 4.28(dd, J=5, 12Hz, 1H), 4.41(dd, J=4, 12Hz, 1H), 5.14(m, 1H), 7.31-7.41(m, 6H), 7.52-7.65(m, 4 H);13 C NMR (CDCl3, 100MHz): δ173.12, 172.22, 170.11, 135.89, 135.63, 133.09, 129.82, 127.35, 72.04, 62.8 5, 34.97, 34.76, 32.55, 30.37, 30.14, 30.04, 29.93, 29.76, 29.53, 27.46, 25.49, 25.36, 19.35, 14.41.

[0142] 1-3-3,3-Hydroxypropane-1,2-diyl distearate (57)

[0143] Compound 56 (protected glycerol; 200 mg, 1 equiv.) was placed in a reaction vessel, which was then filled with nitrogen gas and sealed with a rubber stopper. Methanol (30 mL) was then added via cannula to dissolve the compound, followed by K2CO3 (83 mg, 2 equiv.) and stirring under nitrogen gas at room temperature for 4 hours. Upon completion of the reaction, the reaction mixture was distilled under reduced pressure to remove the solvent. The resulting mixture was dissolved in DCM (50 mL) and distilled water (50 mL) and then placed in a separatory funnel to separate the organic and aqueous layers. The aqueous layer was extracted with DCM (2 × 20 mL), and both organic layers were combined and washed with saturated aqueous NaCl (20 mL). The separated organic layer was removed with anhydrous MgSO4 and filtered. The filtrate was distilled under reduced pressure to remove DCM, and the remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 2:8->5:5) to give compound 57 (131 mg, 70%) as a clear liquid.

[0144] 1 H NMR (CDCl3, 400MHz): δ0.88(t, J=6Hz, 6H), 1.05(m, 9H), 1.26(m, 56H), 1.63(m, 4H), 2.25(m, 4H), 3.77(m, 2H), 4.22(dd, J=5, 12Hz, 1H), 4.42(dd, J=4, 12Hz, 1H), 5.18(m, 1H), 7.31-7.42(m, 6H), 7.65-7.68(m, 4H); 13C NMR (CDCl3, 100MHz): δ173.25, 172.77, 135.83, 135.63, 133.09, 129.82, 127.35, 72.04, 62.85, 34 .97, 34.76, 32.55, 30.37, 30.14, 30.04, 29.93, 29.76, 29.53, 27.46, 25.49, 25.36, 19.35, 14.41.

[0145] [ka]

[0146] 1-4-1,3-((3-((R)-2,2,5,5-Tetramethyl-1,3-dioxane-4-carboxamido)propanoyl)oxy)propane-1,2-diyl distearate (58)

[0147] Compound 52 (186 mg, 1.5 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (111 mg, 1.5 equiv.) and DMAP (12 mg, 0.2 equiv.) were added, and the reaction mixture was vigorously stirred under argon at 5 °C for 10 min. Compound 57 (300 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and stirred at room temperature for 9 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 3:7), the reaction mixture was further dissolved in DCM (50 mL) and transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 x 50 mL) and saturated aqueous NaCl (30 mL). The organic layer was filtered with anhydrous MgSO4 to remove water, and the filtrate was distilled under reduced pressure and purified by flash column chromatography (SiO2, EtOAc / hexane 4:6) to give compound 58 (306 mg, 72%) as a clear oil.

[0148] 1H NMR (CDCl3, 400MHz): δ0.86(t, J=11.1Hz, 6H), 0.95(s, 3H), 1.02(s, 3H), 1.24-1 .31(m, 58H), 1.45(s, 3H), 1.56(s, 3H), 1.57-1.69(m, 4H), 2.28-2.31(q, J=6.2H z, 4H), 2.57(t, J=6.5Hz, 2H), 3.27(d, J=11.5Hz, 1H), 3.44-3.57(m, 2H), 3.68(d , J=11.5Hz, 1H), 4.11(s, 1H), 4.12-4.33(m, 4H), 5.23-5.26(m, 1H), 7.27(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.01, 18.57, 18.58, 18.73, 21.99, 22.58, 24.75, 24.78, 28.97, 29.02, 29.17, 29.26, 29.35, 29.38, 29.39, 29.52, 29.56, 2 9.60, 31.81, 39.82, 32.86, 33.79, 33.81, 33.93, 34.03, 34.07, 61.87, 62 .51, 68.63, 68.64, 71.37, 77.03, 98.92, 169.71, 171.54, 172.79, 173.14.

[0149] 1-4-2,3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl distearate (Compound 11)

[0150] Compound 58 (200 mg, 1 equiv.) and DL-1,4-dithiothreitol (DTT, 71 mg, 2 equiv.) were dissolved in DCM in a reaction vessel by stirring, and then p-toluenesulfonic acid (20 mg, 0.5 equiv.) was added at room temperature and stirred for 1 hour. After confirming completion of the reaction by TLC, the mixture was transferred to a separatory funnel, extracted with EtOAc (30 mL x 2), and washed with saturated aqueous NaCl. The organic layer was removed with anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure. It was then purified by flash column chromatography (SiO2, EtOAc / hexane 2:8->6:4) to give compound 11 (105 mg, 55%) as a yellow oil.

[0151] 1H NMR (CDCl3, 400MHz): δ0.87 (t, J=10.2Hz, 6H), 0.90 (s, 3H), 1.03 (s, 3H), 1.2 5-1.31(m, 59H), 1.60-1.62(m, 4H), 2.30-2.35(q, J=6.3Hz, 4H), 2.54-2.62( m, 2H), 3.48-3.49(m, 1H), 3.52-3.55(m, 3H), 3.59-3.62(m, 1H), 3.91-4.01( m, 1H), 4.04-4.06 (m, 1H), 4.14-4.41 (m, 4H), 5.27-5.33 (m, 1H), 7.27 (s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.11, 20.00, 20.23, 21.64, 21.77, 22.69, 24.85, 24.9 0, 29.06, 29.12, 29.27, 29.36, 29.49, 29.64, 29.67, 29.71, 31.93, 33.99, 34. 02, 34.06, 34.22, 34.50, 39.36, 62.01, 62.07, 62.64, 62.71, 68.82, 68.86, 71.15, 71.19, 77.72, 171.79, 171.85, 173.22, 173.33, 173.48, 173.56, 173.68.

[0152] [Example 2] Synthesis of パントテン acid-based lipids

[0153]

change

[0154] 2-1,9-Hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl dioleate [9-Hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl dioleate] (Compound 12)

[0155] 3-(dimethylamino)propanoic acid (compound 59; 26 mg, 1.2 equiv.) was dissolved in DCM (20 mL), and EDCI·HCl (35 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were added. The mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 10 (100 mg, 1 equiv.) in DCM (5 mL) was then added dropwise at 5 °C and stirred at room temperature for 16 h. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1), an additional 50 mL of DCM was added to the reaction mixture, and the organic layer was washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure and purified by column chromatography (SiO2, EtOAc / MeOH 1:0->9:1) to give compound 12 (37 mg, 37%) as a clear oil.

[0156] 1 H NMR (CDCl3, 400MHz): δ0.87-0.89(m, 9H), 1.18(s, 3H), 1.30-1.41(m, 40H), 1.60-1.61(m, 4H), 1.99-2.03(m, 8H), 2.31(s, 6H) ), 2.32-2.34(m, 4H), 2.52-2.72(m, 7H), 3.52-3.61(m, 3H), 3.90(s, 1H), 4.14-4.33(m, 5H), 5.26-5.39(m, 5H), 7.28(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.06, 14.10, 19.49, 22.29, 22.67, 24.82, 24.85, 27.16, 27.21, 29.97, 29.08, 29.11, 29.17, 29.30, 29.48, 29.51, 29.58, 29.64, 29.70, 29.75, 31.89, 32 .55, 32.59, 33.23, 34.01, 34.15, 34.47, 34.49, 38.44, 38.47, 45.03, 55.66, 62.01, 62.55, 62.59, 68.76, 71.37, 74.49, 74.55, 129.68, 130.01, 130.15, 171.57, 172.09, 172.69.

[0157] [ka]

[0158] 2-2,3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(pyrrolidin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl dioleate] (Compound 13)

[0159] 3-(pyrrolidin-1-yl)propanoic acid (compound 60, 21 mg, 1.2 equiv.) was dissolved in DCM (20 mL) and EDCI·HCl (28 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were added to a reaction vessel. The mixture was vigorously stirred under argon at 5 °C for 10 min. Compound 10 (100 mg, 1 equiv.) in DCM (5 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 12 h. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1), an additional 50 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure and purified by column chromatography (SiO2, EtOAc / MeOH 1:0->9:1) to give compound 13 (47 mg, 41%) as a clear oil.

[0160] 11H NMR (CDCl3, 400 MHz): δ 0.77 - 0.82 (m, 9H), 1.15 (s, 3H), 1.23 - 1.41 (m, 40H), 1.52 - 1.55 (m, 4H), 1.88 (m, 4H), 1.89 - 1.94 (m, 8H), 2.22 - 2.26 (m, 4H), 2.24 - 2.53 (m, 10H), 2.64 - 2.66 (m, 1H), 3.42 - 3.46 (m, 3H), 3.83 (s, 1H), 4.05 - 4.09 (m, 2H), 4.20 - 4.24 (m, 2H), 4.31 - 4.33 (m, 1H), 5.25 - 5.31 (m, 5H), 7.53 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.10, 14.10, 19.04, 22.67, 22.83, 23.12, 24.83, 24.83, 24.85, 27.16, 27.21, 28.97, 29.03, 29.08, 29.11, 29.17, 29.31, 29.48, 29.51, 29.58, 29.65, 29.70, 29.75, 31.89, 32.55, 32.60, 34.01, 34.15, 34.32, 34.40, 38.47, 38.49, 52.77, 52.81, 54.11, 62.01, 62.54, 62.60, 68.75,​​​​​​​​​​​​​​​(9Z,12Z)-octadeca-9,12-dienoic acid (710 mg, 2.5 equiv.) was dissolved in DCM (40 mL) and then EDCI·HCl (578 mg, 2.5 equiv.) and DMAP (36 mg, 0.2 equiv.) were added to a reaction vessel. The mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 55 (200 mg, 1 equiv.) was then added dropwise to DCM (20 mL) at 5 °C and stirred at room temperature for 20 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 1:1), an additional 50 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure and purified by column chromatography (SiO2, EtOAc / hexane 2:8) to give glycerol compound 61 (845 mg, 86%) as a clear oil.

[0164] 1 H NMR (CDCl3, 400MHz): δ0.89(t, J=6.9Hz, 6H), 1.40-1.26(m, 28H), 1.63(m, 4H), 2.05(dt, J=6.6, 6.9Hz, 8H), 2.11(s, 3H), 2.33(t, J=7.8Hz, 2H), 2.36(t, J=7.8Hz, 2H), 2.79(t, J=6 .5Hz, 4H), 3.74(s, 3H), 4.23(dd, J=5.8, 11.7Hz, 1H), 4.33(dd, J=4.7, 11.8Hz, 1H), 4.44( d, J=11.8Hz, 1H), 4.48(d, J=11.8Hz, 1H), 5.06(tt, J=5.6, 5.6Hz, 1H), 5.39-5.31(m, 8H); 13C NMR (CDCl3, 100MHz): δ14.08(2C), 20.83, 22.58(2C), 24.87(2C), 24.92(2C), 25.62(2C), 27.18(4C), 29.06(2C), 29.13(2C), 29.18(2C), 2 9.36(2C), 29.61(2C), 31.53(2C), 34.06(2C), 34.25(2C), 61.54, 61. 99, 72.11, 127.89, 128.08, 130.01, 130.21, 173.39, 173.76, 175.87.

[0165] 2-3-2,3-Hydroxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) [3-Hydroxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate)] (62)

[0166] Glycerol compound 61 (845, 1 equiv.) was placed in a reaction vessel, sealed with a rubber stopper, and then flushed with nitrogen gas until the reaction was complete. CHOH (70 mL) was then added via cannula, followed by KCO (354 mg, 2 equiv.) and stirring at room temperature for 7 hours. The reaction mixture was distilled under reduced pressure to remove the solvent, followed by the addition of DCM (50 mL) and distilled water (50 mL). The resulting solution was transferred to a separatory funnel. The aqueous layer was extracted with DCM (2 × 20 mL), and both organic layers were combined and washed with saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO and filtered. The filtrate was distilled under reduced pressure and purified by column chromatography (SiO, 1:9->5:5 EtOAc / hexane) to give compound 62 (561 mg, 71%) as a clear oil.

[0167] 11H NMR (CDCl3, 400 MHz): δ 0.89 (t, J = 6.9 Hz, 6H), 1.26 - 1.40 (m, 28H), 1.63 (m, 4H), 2.05 (dt, J = 6.6, 6.9 Hz, 8H), 2.33 (t, J = 7.8 Hz, 2H), 2.36 (t, J = 7.8 Hz, 2H), 2.79 (t, J = 6.5 Hz, 4H), 3.74 (s, 3H), 4.23 (dd, J = 5.8, 11.7 Hz, 1H), 4.33 (dd, J = 4.7, 11.8 Hz, 1H), 4.44 (d, J = 11.8 Hz, 1H), 4.48 (d, J = 11.8 Hz, 1H), 5.06 (tt, J = 5.6, 5.6 Hz, 1H), 5.31 - 5.39 (m, 8H); 13 13C NMR (CDCl3, 100 MHz): δ 14.08 (2C), 22.58 (2C), 24.87 (2C), 24.92 (2C), 25.62 (2C), 27.18 (4C), 29.06 (2C), 29.13 (2C), 29.18 (2C), 29.36 (2C), 29.61 (2C), 31.53 (2C), 34.06 (2C), 34.25 (2C), 61.54, 61.99, 72.11, 127.89, 128.08, 130.01, 130.21, 173.39, 173.76。

[0168]

Chem.

[0171] 1 H NMR (CDCl3, 400MHz): δ0.90(t, J=6.8Hz, 6H), 0.98(s, 3H), 1.05(s, 3H), 1.28 -1.35(m, 29H), 1.43(s, 3H), 1.47(s, 3H), 1.61-1.64(m, 4H), 2.07(dd, J=6.4 , 10.1Hz, 8H), 2.41(m, 4H), 2.61(t, J=6.7, 3H), 2.78(t, J=5.8Hz, 3H), 3.27- 3.71(m, 4H), 4.09(s, 1H), 4.17-4.34(m, 4H), 5.27-5.41(m, 9H), 7.01(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.06, 18.69, 18.83, 22.10, 22.56, 24.82, 24.85, 25.63, 27.19, 29.03, 29.08, 29 .11, 29.17, 29.34, 29.45, 29.60, 31.52, 32.52, 33.90, 34.00, 34.14, 37.18, 61.98, 62.55, 62.60, 68.76. 71.47, 99.02, 127.89, 128.08, 129.97, 130.21, 169.81, 171.63, 172.81, 173.81.

[0172] 2-4-2,3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) [3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate)] (64)

[0173] Compound 63 (580 mg, 1 equiv.) and DTT (208 mg, 2 equiv.) were mixed in a reaction vessel with DCM, followed by the addition of p-toluenesulfonic acid (78 mg, 0.5 equiv.) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes, and after confirming completion of the reaction by TLC, it was extracted with EtOAc (30 mL x 2) and the organic layer was washed with saturated aqueous NaCl. The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 3:7->7:3) to give the diol compound 64 (408 mg, 55%) as a pale yellow oil.

[0174] 1 H NMR (CDCl3, 400MHz): δ0.91(t, J=6.8Hz, 6H), 0.94(s, 3H), 1.05(s, 3H), 1.2 7-1.41(m, 29H), 1.61-1.63(m, 4H), 2.07(dd, J=6.5, 10.0Hz, 8H), 2.37-2.43 (m, 4H), 2.58-2.77(m, 2H), 2.79-2.80(t, J=5.7Hz, 4H), 3.25(m, 2H), 3.50-3 .55(m, 4H), 4.03(s, 1H), 4.04-4.31(m, 4H), 5.31-5.42(m, 9H), 7.92(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.07, 20.00, 20.23, 21.63, 21.75, 22.57, 24.82, 24.87, 25.64, 27.16, 27.20, 29.06, 29.08, 29.12, 29.17, 29.34, 29.61, 31.35, 33.99, 34.99, 34.19, 34.52, 39.35, 3 9.36, 62.01, 62.06, 62.62, 62.70, 68.83, 68.68, 71.16, 71.19, 127.89, 128.10, 128.11, 129.97, 129.98, 130.24, 171.77, 171.83, 173.83, 173.16, 173.32, 173.43, 173.46, 173.50, 173.62.

[0175] 2-4-3. (9R)-9-Hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) [(9R)-9-Hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate)] (Compound 14)

[0176] Compound 59 (34 mg, 1.2 equiv.) was dissolved in DCM (20 mL) and EDCI·HCl (210 mg, 1.5 equiv.) and DMAP (22 mg, 0.2 equiv.) were added to the reaction vessel. The mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 64 (200 mg, 1 equiv.) was then dissolved in DCM (5 mL) and added dropwise at 5 °C. The mixture was then stirred at room temperature for 14 h. After TLC (SiO2, EtOAc / MeOH 9:1) confirmed the reaction was complete, an additional 50 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 1:0->9:1) to give compound 14 (241 mg, 29%) as a clear oil.

[0177] 11H NMR (CDCl3, 400 MHz): δ 0.87 - 0.90 (m, 9H), 0.71 (s, 3H), 1.25 - 1.37 (m, 30H), 1.59 - 1.62 (m, 4H), 2.05 (dd, J = 6.5, 10.0 Hz, 8H), 2.26 (s, 6H), 2.30 - 2.34 (m, 4H), 2.52 - 2.65 (m, 6H), 2.76 (t, J = 5.7 Hz, 4H), 3.52 - 3.61 (m, 3H), 3.90 (s, 1H), 4.12 - 4.31 (m, 6H), 5.30 - 5.39 (m, 9H), 7.41 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.05, 19.47, 22.22, 22.55, 24.81, 24.83, 25.61, 27.17, 29.01, 29.06, 20.10, 19.16, 29.32, 29.59, 31.50, 33.09, 33.99, 34.13, 34.47, 38.42, 38.44, 44.95, 55.50, 62.01, 62.54, 62.85, 68.76, 71.33, 74.43, 74.48, 127.87, 124.06, 129.97, 130.19, 171,55, 171.55, 172.04, 172.78, 172.89, 172.94, 173.24, 173.26。

[0178] [Chemical formula]

[0179] 2-5-1,3-Acetoxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate) [3-Acetoxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate)] (65) <000105o> (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid (linolenic acid; 520 mg, 2.5 equiv.) was dissolved in DCM (40 mL) in a reaction vessel, followed by the addition of EDCI·HCl (290 mg, 2.5 equiv.) and DMAP (18 mg, 0.2 equiv.) and vigorously stirring under argon gas at 5°C for 10 minutes. Compound 55 (100 mg, 1 equiv.) dissolved in DCM (20 mL) was then added dropwise at 5°C and the reaction mixture was stirred at room temperature for 25 hours. After confirming the completion of the reaction by TLC (SiO2, EtOAc / hexane 1:1), additional DCM (50 mL) was added and washed with saturated aqueous NaHCO3 (2 x 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 3:7) to give compound 65 (391 mg, 80%) as a clear oil.

[0181] 1 H NMR (CDCl3, 400MHz): δ0.91(t, J=6.3Hz, 6H), 1.25-1.41(m, 20H), 1.60-1.62(m, 5H), 2.04-2.14(m, 8H), 2.11(s, 3 H), 2.31-2.37(m, 4H), 2.84-2.88(m, 8H), 4.12(s, 1H), 4.32-4.38(m, 4H), 5.27-5.33(m, 1H), 5.34-5.43(m, 12H); 13 C NMR (CDCl3, 100MHz): δ14.06(2C), 21.57(2C), 24.88(2C), 24.97(2C), 26.62(2C), 27.16(4C), 29.06(2C), 30.13(2C), 29.11(2C), 29.3 5(2C), 29.61(2C), 31.55(2C), 34.06(2C), 34.27(2C), 61.53, 61.98, 72.12, 127.90, 128.08, 130.02, 130.22, 173.40, 173.77, 175.88.

[0182] 2-5-2,3-Hydroxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate) [3-Hydroxypropane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate)] (66)

[0183] Compound 65 (390, 1 equiv.) was placed in a reaction vessel, sealed with a rubber stopper, and then filled with nitrogen gas until the reaction was complete. CHOH (70 mL) was then added via cannula, followed by KCO (164 mg, 2 equiv.) and stirred at room temperature for 9 hours. DCM (50 mL) and distilled water (50 mL) were added to the reaction mixture, and the resulting solution was transferred to a separatory funnel. The aqueous layer was extracted with DCM (2 × 20 mL), and both organic layers were combined and washed with saturated aqueous NaCl (20 mL). The resulting organic layer was dried over anhydrous MgSO and filtered. The filtrate was evaporated under reduced pressure and purified by flash column chromatography (SiO, EtOAc / hexane 2:8->5:5) to give compound 66 (222 mg, 61%) as a clear oil.

[0184] 1 H NMR (CDCl3, 400MHz): δ0.90(t, J=6.9Hz, 6H), 1.26-1.41(m, 20H), 1.61-1.63(m, 5H), 2.04-2.13(m, 8H), 2. 31-2.35(m, 4H), 2.84-2.89(m, 8H), 4.10(s, 1H), 4.32-4.39(m, 4H), 5.27-5.32(m, 1H), 5.34-5.42(m, 12H); 13 C NMR (CDCl3, 100MHz): δ14.06(2C), 21.57(2C), 24.88(2C), 24.97(2C), 26.62(2C), 27.16(4C), 29.06(2C), 30.13(2C), 29.11(2C), 29.35(2C), 29.61(2C), 31.55(2C), 34.06(2C), 34.27(2C), 61.53, 61.98, 72.12, 127.90, 128.08, 130.02, 130.22, 173.40, 173.77.

[0185] [ka]

[0186] 2-6-1,3-((3-((R)-2,2,5,5-Tetramethyl-1,3-dioxane-4-carboxamido)propanoyl)oxy)propane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate) (67)

[0187] Compound 55 (141 mg, 1.5 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (82 mg, 1.5 equiv.) and DMAP (9 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 66 (222 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 12 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 3:7), additional DCM (50 mL) was added, the mixture was transferred to a separatory funnel, and washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 2:8) to give compound 67 (232 mg, 75%) as a clear oil.

[0188] 1 H NMR (CDCl3, 400MHz): δ0.97-1.01(m, 9H), 1.06(m, 3H), 1.32-1.33(m, 20H), 1.44(m, 3H), 1.4 8(m, 3H), 1.61-1.63(m, 5H), 1.73(s, 1H), 2.04-2.13(m, 8H), 2.31-2.35(m, 4H), 2.58-2.61(t , J=6.9Hz, 2H), 2.81-2.84(m, 8H), 3.32(d, J=11.6Hz, 1H), 3.41-3.68(m, 2H), 3.75(d, J=11.8 Hz, 1H), 4.10(s, 1H), 4.15-4.34(m, 4H), 5.27-5.32(m, 1H), 5.34-5.42(m, 12H), 7.29(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.12, 18.84, 20.55, 22.10, 24.82, 24.84, 25.52, 25.61, 27. 20, 29.03, 29.08, 29.11, 29.17, 29.46, 29.59, 32.97, 33.91, 34.00, 34.14, 34.18, 61 .98, 61.55, 62.60, 68.75, 71.47, 76.72, 77.04, 77.15, 77.36, 99.03, 127.11, 127.76, 127.77, 128.23, 129.30, 130.20, 130.21, 131.95, 168.82, 171.64, 172.82, 173.20.

[0189] 2-6-2,3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate) (68)

[0190] Compound 67 (230 mg, 1 equiv.) and DTT (83 mg, 2 equiv.) were dissolved in DCM, followed by the addition of p-toluenesulfonic acid (23 mg, 0.5 equiv.) at room temperature and stirring for 30 minutes. After the reaction was completed, the reaction mixture was extracted with EtOAc (30 mL x 2). The combined organic layers were washed with saturated aqueous NaCl. The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 3:7->5:5) to give compound 68 (121 mg, 55%) as a pale yellow oil.

[0191] 1 H NMR (CDCl3, 400MHz): δ0.98-1.02(m, 9H), 1.03(m, 3H), 1.32-1.34(m, 20H), 1.61-1. 65(m, 5H), 1.72(s, 1H), 2.04-2.15(m, 8H), 2.34-2.37(m, 4H), 2.59-2.65(t, J=6.9H z, 2H), 2.81-2.84(m, 8H), 3.32-3.33(m, 1H), 3.41-3.69(m, 2H), 3.75-3.78(m, 1H), 4.10(s, 1H), 4.16-4.34(m, 4H), 5.28-5.32(m, 1H), 5.34-5.42(m, 12H), 7.30(s, 1H);13 C NMR (CDCl3, 100MHz): δ14.13, 18.85, 24.83, 24.85, 25.53, 25.63, 27.23, 29.04, 29.07, 29.12, 29.18, 29.45, 29.60, 32.98, 33.91, 34.11, 34.12, 34.12, 61.98, 61 .56, 62.61, 68.76, 71.48, 76.71, 77.03, 77.16, 77.36, 99.04, 127.12, 127.74, 127.77, 128.24, 129.30, 130.21, 130.25, 131.94, 168.82, 171.64, 172.82, 173.20.

[0192] 2-6-3. (9R)-9-hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate) [(9R)-9-hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl(9Z,9'Z,12Z,12'Z,15Z,15'Z)-bis(octadeca-9,12,15-trienoate)] (Compound 15)

[0193] Compound 59 (31 mg, 1.2 equiv.) was dissolved in DCM (20 mL) in a reaction vessel. EDCI·HCl (34 mg, 1.5 equiv.) and DMAP (3.6 mg, 0.2 equiv.) were added to the reaction mixture and vigorously stirred at 5 °C for 10 min under argon gas. Compound 68 (120 mg, 1 equiv.) in DCM (5 mL) was then added dropwise at 5 °C and the reaction mixture was stirred at room temperature for 18 h. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1), additional DCM (50 mL) was added, the mixture was transferred to a separatory funnel, and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 1:0->9:1) to give compound 15 (48 mg, 36%) as a clear oil.

[0194] 11H NMR (CDCl3, 400 MHz): δ 0.89 (s, 3H), 0.97 (t, J = 7.6 Hz, 6H), 1.09 (s, 3H), 1.25 - 1.30 (m, 20H), 1.60 (s, 5H), 2.02 - 2.11 (m, 8H), 2.29 - 2.34 (m, 4H), 2.57 (s, 2H), 2.79 - 3.01 (m, 16H), 3.35 - 3.36 (m, 4H), 3.73 - 3.76 (m, 1H), 4.05 (s, 1H), 4.11 - 4.17 (m, 3H), 4.25 - 4.31 (m, 3H), 5.27 (m, 1H), 5.30 - 5.36 (m, 12H), 7.41 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.26, 19.48, 20.53, 21.71, 24.80, 24.84, 24.71, 24.80, 24.84, 25.51, 25.60, 27.18, 29.02, 29.06, 29.10, 29.16, 29.54, 30.27, 34.00, 34.15, 34.49, 38.17, 43.79, 53.76, 62.05, 62.56, 68.78, 71.62, 73.65, 73.73, 127.09, 127.73, 128.21, 128.27, 130.20, 131.93, 169.91, 171.57, 173.01, 173.32。

[0195]

Chemistry

[0196] 2-7-1,3-Acetoxypropane-1,2-diyl bis(2-hexyldecanoate) [3-Acetoxypropane-1,2-diyl bis(2-hexyldecanoate)] (69)

[0197] 2-Hexyldecanoic acid (956 mg, 2.5 equiv.) was placed in a reaction vessel and dissolved in DCM (40 mL). EDCI·HCl (580 mg, 2.5 equiv.) and DMAP (55 mg, 0.3 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 55 (200 mg, 1 equiv.) in DCM (20 mL) was then added dropwise to the reaction mixture at 5 °C and stirred at room temperature for 24 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 4:6), an additional 50 mL of DCM was added, transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9) to give compound 69 (801 mg, 88%) as a clear liquid.

[0198] 1 H NMR (CDCl3, 400MHz): δ0.81-0.82(t, J=5.2Hz, 12H), 1.18-1.25(m, 42H), 1. 36-1.41(m, 5H), 1.49-1.55(m, 5H), 2.11(s, 3H), 2.25-2.31(m, 2H), 3.54-3 .57(dd, J=3.6, 3.9Hz, 1H), 3.60-3.64(dd, J=3.6, 3.9Hz, 1H), 4.10-4.15(d d, J=3.4, 4.3Hz, 1H), 4.28-4.32(dd, J=3.4, 4.3Hz, 1H), 5.12-5.16(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.04, 14.08, 20.83, 22.61, 22.66, 27.31, 27.34, 27.31, 27.42, 27.46, 29.20, 29.23, 29.2 8, 29.45, 29.53, 29.56, 31.68, 32.31, 32.34, 32.19, 42.32, 45.65, 45.74, 62.09, 70.17, 171.56, 175.46, 175.87.

[0199] 2-7-2,3-Hydroxypropane-1,2-diylbis(2-hexyldecanoate) [3-Hydroxypropane-1,2-diyl bis(2-hexyldecanoate)] (70)

[0200] Glycerol compound 69 (700 mg, 1 equiv.) was placed in a reaction vessel, sealed with a rubber stopper, and then filled with nitrogen gas until the reaction was complete. CHOH (70 mL) was then added via cannula, followed by KCO (316 mg, 2 equiv.) and stirred at room temperature for 6 hours. The reaction mixture was evaporated under reduced pressure to remove the solvent, and then DCM (50 mL) and distilled water (50 mL) were added and transferred to a separatory funnel. The aqueous layer was extracted with DCM (2 × 20 mL). Both organic layers were combined and washed with saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO and filtered. The filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO, EtOAc / hexane 2:8->3:6) to give compound 70 (502 mg, 77%) as a clear oil.

[0201] 1 H NMR (CDCl3, 400MHz): δ0.80-0.82(t, J=5.1Hz, 12H), 1.18-1.24(m, 42H), 1.36-1.41(m, 5H), 1.49-1.57(m, 5H), 2.25-2.31(m, 2H), 3.54-3.58(d d, J=3.6, 3.9Hz, 1H), 3.60-3.64(dd, J=3.6, 3.9Hz, 1H), 4.10-4.14(dd, J=3.4, 4.3Hz, 1H), 4.28-4.32(dd, J=3.4, 4.3Hz, 1H), 5.12-5.16(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.04, 14.08, 22.60, 22.66, 27.30, 27.34, 27.37, 27.41, 27.45, 29.19, 29.23, 29. 28, 29.44, 29.53, 29.58, 31.68, 32.30, 32.33, 32.18, 42.32, 45.64, 45.73, 62.08, 70.16, 175.46, 175.86.

[0202] [ka]

[0203] 2-8-1,3-((3-((R)-2,2,5,5-Tetramethyl-1,3-dioxane-4-carboxamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) (71)

[0204] Acetal-protected pantothenic acid compound 52 (376 mg, 1.5 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (225 mg, 1.5 equiv.) and DMAP (24 mg, 0.2 equiv.) were then added and vigorously stirred under argon gas at 5 °C for 10 min. Compound 70 (550 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the reaction mixture was stirred at room temperature for 11 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 2:8), additional DCM (50 mL) was added, the mixture was transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was distilled under reduced pressure to remove the solvent, and the resulting mixture was purified by column chromatography (SiO2, EtOAc / hexane 3:7) to obtain compound 71 (440 mg, 77%) as a clear oil.

[0205] 1 H NMR (CDCl3, 400MHz): δ0.61-0.63(t, J=5.1Hz, 12H), 0.73(s, 3H), 0.80(s, 3H), 1.0 1-1.07(m, 41H), 1.18-1.22(m, 11H), 1.32-1.37(m, 5H), 2.08-2.11(m, 2H), 2.30-2. 33(t, J=4.1Hz, 2H), 3.02-3.05(d, J=6.1Hz, 1H), 3.20-3.42(m, 2H), 3.43-3.45(d, J =6.1Hz, 1H), 3.84-3.94(m, 3H), 4.06-4.31(m, 2H), 5.04-5.06(m, 1H), 7.04(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.03, 14.07, 18.66, 22.08, 22.58, 22.64, 27.26, 27.32, 27.37, 27.42, 29.18, 29.21, 29.27, 29.42, 29.54, 29.57, 31.65, 3 1.84, 32.21, 32.27, 32.95, 33.83, 33.85, 34.09, 32.12, 45.57, 45.63, 61 .95, 62.72, 62.79, 68.64, 71.46, 99.00, 169.78, 171.55, 175.49, 175.87.

[0206] 2-8-2,3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2,4-Dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate)] (72)

[0207] Compound 71 (420 mg, 1 equiv.) and DTT (160 mg, 2 equiv.) were placed in a reaction vessel and dissolved in DCM. p-Toluenesulfonic acid (45 mg, 0.5 equiv.) was added at room temperature and stirred for 1 hour. Upon completion of the reaction, the mixture was extracted with EtOAc (30 mL x 2), and the combined organic layers were washed with saturated aqueous NaCl. The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 3:7->6:4) to give compound 72 (243 mg, 61%) as a pale yellow oil.

[0208] 1 H NMR (CDCl3, 400MHz): δ0.79-0.82(t, J=5.1Hz, 12H), 0.95(s, 3H), 1.00(s, 3H), 1.18-1.24(m, 45H), 1.38(m, 5H), 1.48-1.52(m, 5H), 2.25 -2.29(m, 2H), 2.43-2.47(m, 2H), 3.31-3.43(m, 1H), 3.59-3.62(m, 1H), 4.02-4.06(m, 3H), 4.15-4.28(m, 4H), 4.38-4.42(m, 1H), 5.32-. 5.34(m, 1H), 7.02-7.20(m, 1H); 13C NMR (CDCl3, 100MHz): δ14.03, 14.07, 19.91, 19.98, 20.84, 22.59, 22.65, 27.23, 27.28, 2 7.33, 27.38, 27.44, 29.11, 29.20, 29.27, 29.43, 29.56, 31.62, 31.65, 31.85, 32.21, 32.2 6, 32.28, 32.32, 32.43, 33.88, 34.41, 34.61, 38.29, 38.52, 45.62, 45.74, 61.95, 62.08, 62.80, 63.03, 68.70, 68.76, 73.27, 73.32, 74.70, 74.96, 169.66, 172.05, 173.54, 173.71.

[0209] 2-8-3. (9R)-9-Hydroxy-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazaoctadecane-17,18-diyl bis(2-hexyldecanoate) (Compound 16)

[0210] Compound 59 (37 mg, 1.2 equiv.) was dissolved in DCM (20 mL) in a reaction vessel. EDCI·HCl (60 mg, 1.5 equiv.) and DMAP (6.3 mg, 0.2 equiv.) were added and vigorously stirred under argon gas at 5 °C for 10 min. Compound 72 (200 mg, 1 equiv.) in DCM (5 mL) was then added dropwise at 5 °C and the reaction mixture was stirred at room temperature for 15 h. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1), additional DCM (50 mL) was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 1:0->9:1, with 28% aqueous NH3) to give compound 16 (104 mg, 46%) as a clear oil.

[0211] 11H NMR (CDCl3, 400 MHz): δ 0.79 - 0.80 (m, 12H), 0.81 (s, 3H), 1.11 (s, 3H), 1.18 - 1.22 (m, 42H), 1.36 - 1.39 (m, 4H), 1.48 - 1.51 (m, 4H), 2.21 - 2.26 (m, 8H), 2.43 - 2.55 (m, 4H), 2.56 - 2.67 (m, 2H), 3.43 - 3.54 (m, 3H), 3.83 (s, 1H), 3.99 - 4.05 (m, 2H), 4.22 - 4.30 (m, 3H), 5.21 - 5.23 (m, 1H), 7.39 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.03, 14.07, 19.91, 19.98, 20.84, 22.59, 22.65, 27.23, 27.28, 27.33, 27.38, 27.44, 29.11, 29.20, 29.27, 29.43, 29.56, 31.62, 31.65, 31.85, 32.21, 32.26, 32.28, 32.32, 32.43, 33.88, 34.41, 34.61, 38.29, 38.52, 45.62, 45.74, 61.95, 62.08, 62.80, 63.03, 68.70, 68.76, 73.27, 73.32, 74.70, 74.96, 169.66, 172.05, 173.54, 173.71。

[0212] [Chemical formula]

[0213] 2-9. 3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(pyrrolidin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(pyrrolidin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 17)

[0214] Compound 60 (41 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DMF (20 mL). EDCI·HCl (75 mg, 1.5 equiv.) and DMAP (6.3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes under argon gas. Compound 72 (200 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DMF (10 mL). The mixture was cooled to 5 °C and the mixture of compound 60 was added dropwise at 5 °C for 30 minutes, followed by stirring at 100 °C for 27 minutes. After TLC (SiO2, EtOAc / MeOH 1:1 and 9:1) confirmed the reaction was complete, the reaction mixture was evaporated under reduced pressure to remove the solvent, and DCM (50 mL) was added. The mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, DCM / MeOH 10:0->9:1, with 28% aqueous NH3) to give compound 17 (151 mg, 65%) as a clear oil.

[0215] 1 H NMR (CDCl3, 400MHz): δ0.77-0.82(m, 16H), 1.15-1.22(m, 46H), 1.34-1.3 9(m, 4H), 1.48-1.51(m, 4H), 1.72(s, 4H), 2.25-2.27(m, 2H), 2.44-2.57( m, 8H), 2.67-2.68(m, 1H), 2.81-2.91(m, 1H), 3.43-3.48(m, 3H), 3.83(s, 1H), 4.01-4.06(m, 2H), 4.22-4.34(m, 3H), 5.20-5.23(m, 1H), 7.3(m, 1H); 13C NMR (CDCl3, 100MHz): δ14.05, 14.09, 19.00, 22.60, 22.65, 22.85, 23.12, 27.26, 27.33, 27.35 ,27.38,27.43,29.19,29.23,29.29,29.44,29.55,29.58,31.66,31.85,32.21,32.27,33.96, 34.28, 38.45, 38.48, 45.58, 45.63, 45.65, 52.72, 52.77, 54.12, 62.01, 62.69, 62.77, 68.64, 71.51, 71.55, 74.45, 74.51, 171.51, 171.55, 172.79, 175.53, 175.63, 175.65, 175.95, 175.98.

[0216]

change

[0217] 2-10. 3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(piperidin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(piperidin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 18)

[0218] 3-(piperidin-1-yl)propanoic acid (compound 73, 40.40 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (67 mg, 1.5 equiv.) and DMAP (5.7 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes. Compound 72 (180 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL) and cooled to 5°C. Compound 73 was then added dropwise at 5°C for 30 minutes and the mixture was stirred at room temperature for 9 hours. After TLC (SiO2, EtOAc / MeOH 1:1 and 9:1) confirmed the reaction was complete, DCM (50 mL) was added, transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash column chromatography (SiO2, DCM / MeOH 10:0 → 8:1, with 28% aqueous NH3) to give compound 18 (130 mg, 61%) as a clear liquid.

[0219] 1 H NMR (CDCl3, 400MHz): δ0.77-0.82(m, 15H), 0.14-1.24(m, 43H), 1.34-1.39(m, 6H), 1.50(s, 8H), 2.24-2.52(m, 10H), 2.71(m, 1H) , 3.42-3.49(m, 3H), 3.83(s, 1H), 4.02-4.06(m, 2H), 4.24-4.35(m, 2H), 4.37-4.38(m, 1H), 5.20-5.22(s, 1H), 7.30-7.34(m, 1H); 13C NMR (CDCl3, 100MHz): δ14.04, 14.08, 18.95, 19.98, 22.60, 22.65, 22.80, 22.84, 23.96, 25.00, 27.26, 2 7.30, 27.32, 27.35, 27.38, 27.43, 29.18, 29.22, 29.28, 29.43, 29.55, 29.58, 31.66, 31.85, 32.51, 32. 27, 32.40, 32.97, 34.23, 38.48, 38.52, 45.58, 45.62, 45.64, 54.44, 55.23, 55.28, 62.00, 62.69, 62.44, 68.63, 71.27, 71.32, 74.20, 74.29, 171.52, 171.57, 172.24, 172.70, 175.56, 175.66, 175.94, 175.98.

[0220]

change

[0221] 2-11. 3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(4-methylpiperazin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(4-methylpiperazin-1-yl)propanoyl)oxy)butanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 19)

[0222] 3-(4-methylpiperazin-1-yl)propanoic acid (compound 74, 50 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (75 mg, 1.5 equiv.) and DMAP (6.3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes. Compound 72 (200 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C and the mixture was added dropwise with compound 74 at 5°C for 30 minutes. The mixture was then stirred at room temperature for 8 hours. After TLC (SiO2, EtOAc / MeOH 1:1 and 9:1, PMA stain) confirmed the reaction was complete, the mixture was transferred to a separatory funnel with additional DCM (50 mL) and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, DCM / MeOH 10:0 → 9:1, with 28% aqueous NH3) to give compound 19 (60 mg, 67%) as a clear oil.

[0223] 1 H NMR (CDCl3, 400MHz): δ0.86-0.90 (m, 15H), 1.75 (d, J=1.4Hz, 3H), 1.26-1. 44(m, 46H), 1.45-1.47(m, 4H), 1.56-1.59(m, 4H), 2.31-2.35(m, 5H), 2.49- 2.71(m, 13H), 2.78-2.83(m, 1H), 3.54-3.62(m, 3H), 3.94(s, 1H), 4.07-4. 17(m, 2H), 4.30-4.38(m, 3H), 4.71(s, 1H), 5.28-5.31(m, 1H), 7.31(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.05, 14.09, 19.09, 19.12, 22.34, 22.60, 22.65, 27.26, 27.32, 27.35, 27 .38, 27.43, 29.18, 29.22, 29.28, 29.43, 29.54, 29.58, 31.66, 31.85, 32.21, 32.31, 32.42, 33.96, 34.30, 38.55, 38.61, 45.58, 45.65, 52.46, 54.20, 54.24, 54.36, 54.39, 62.00, 62.74, 62.80, 68.63, 71.03, 71.08, 74.12, 74.52, 171.58, 171.64, 172.41, 172.61, 175.59, 175.74, 175.98, 176.02.

[0224]

change

[0225] 2-12. 3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-morpholinopropanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-morpholinopropanoyl)oxy)butanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 20)

[0226] 3-morpholinopropanoic acid (compound 75, 45.42 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (75 mg, 1.5 equiv.) and DMAP (6.3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes under argon gas. Compound 72 (200 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C, and the mixture was added dropwise with compound 75 at 5°C for 30 minutes. The mixture was then stirred at room temperature for 10 hours. After TLC (SiO2, EtOAc / MeOH 1:1 and 9:1, PMA stain) confirmed the reaction was complete, DCM (50 mL) was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, DCM-MeOH 10:0 → 10:0.2, with 28% aqueous NH3) to give compound 20 (151 mg, 64%) as a clear liquid.

[0227] 1 H NMR (CDCl3, 400MHz): δ0.79-0.82(m, 15H), 1.09(d, J=2.48Hz, 3H), 1.18-1.24(m, 43H), 1.34-1.39(m, 4H), 1.48-1.51(m, 4H), 2.24-2. 28(m, 2H), 2.41-2.52(m, 10H), 3.48-3.63(m, 7H), 3.85(d, J=6Hz, 1H), 4.00-4.04(m, 2H), 4.23-4.37(m, 4H), 5.23(m, 1H), 7.19(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.06, 14.10, 19.22, 19.27, 22.21, 22.61, 22.66, 27.34, 27.39 ,24.44,29.19,29.24,29.30,29.45,29.59,31.68,31.87,32.22,32.27,33.93,34.33, 34.37, 38.57, 38.65, 45.60, 45.66, 53.45, 54.62, 62.03, 62.80, 62.86, 66.22, 68.65, 71.04, 71.10, 74.42, 74.58, 171.64, 171.72, 172.17, 175.66, 175.84, 176.13, 176.08.

[0228]

change

[0229] 2-13. 3-((3-((R)-4-((3-(1H-Pyrazol-1-yl)propanoyl)oxy)-2-hydroxy-3,3-dimethyl butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-4-((3-(1H-Pyrazol-1-yl)propanoyl)oxy)-2-hydroxy-3,3-dimethyl butanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 21)

[0230] 3-(1H-pyrazol-1-yl)propanoic acid (compound 76, 36.75 mg, 1.01 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (79.65 mg, 1.6 equiv.) and DMAP (6.34 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes under argon gas. Compound 72 (200 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C, and the mixture was added dropwise with compound 76 at 5°C for 30 minutes. The mixture was then stirred at room temperature for 14 hours. After confirming the completion of the reaction by TLC (SiO2, EtOAc / MeOH 1:1 and 9:1, PMA stain), the reaction mixture was evaporated under reduced pressure to remove the solvent, then DCM (50 mL) was added and transferred to a separatory funnel. The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by column chromatography (SiO2, DCM / MeOH 10:0->9:1, with 28% NH3 aqueous solution) to give compound 21 (127 mg, 55%) as a clear oil.

[0231] 1 H NMR (CDCl3, 400MHz): δ0.78-0.80(m, 15H), 0.82(s, 3H), 1.05-1.18(m, 42H), 1.22-1.34(m, 4H), 1.36-1.39(m, 4H), 2.25-2.28(m, 4H), 2.4 1-2.56(m, 3H), 3.44-3.48(m, 3H), 3.84(s, 1H), 4.01-4.06(m, 2H), 4.22-4.35(m, 3H), 5.20-5.23(m, 1H), 7.40(m, 1H), 7.42-7.75(m, 3H); 13C NMR (CDCl3, 100MHz): δ14.05, 14.09, 19.16, 19.23, 20.80, 21.26, 21.87, 22.60, 22.65, 27.26, 27.33, 27.38, 27.43, 29.12, 29.28, 29.44, 29.54, 29.58, 31.66, 31.86, 32.26, 33.62, 33.63, 34.35, 34.40, 34.87, 35.18, 35.88, 35.93, 37.35, 37.36, 3 8.38, 38.46, 47.15, 47.21, 47.77, 62.06, 62.75, 62.80, 68.65, 69.69, 71.24, 74.07, 74.25, 76.73, 77.04, 77.16, 77.36, 105.45, 105.81, 106.84, 129.64, 129.70, 129.73, 139.70, 129.73, 139.70, 167.69, 170.76, 171.61, 172.34, 175.61, 175.92.

[0232]

change

[0233] 2-14. 3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(pyridin-4-yl)propanoyl)oxy)butanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-2-Hydroxy-3,3-dimethyl-4-((3-(pyridin-4-yl)propanoyl)oxy)butanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 22)

[0234] 3-(pyridin-4-yl)propanoic acid (compound 77, 32.38 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (60 mg, 1.6 equiv.) and DMAP (4.75 mg, 0.2 equiv.) were added and stirred at room temperature for 20 minutes under argon gas. Compound 72 (150 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C, and the mixture with compound 77 was added dropwise at 5°C for 30 minutes. The mixture was then stirred at room temperature for 10 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / MeOH 1:1 and 9:1, PMA stain), the reaction mixture was evaporated under reduced pressure to remove the solvent, followed by the addition of DCM (50 mL). The mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by column chromatography (SiO2, DCM / MeOH 10:0 -> 9.5:0.1, with 28% aqueous NH3) to give compound 22 (121 mg, 69%) as a clear oil.

[0235] 1 H NMR (CDCl3, 400MHz): δ0.78-0.84(m, 15H), 0.94(s, 3H)1.18-1.23(m, 42H), 1 .36-1.37(m, 4H), 1.49-1.50(m, 4H), 2.24-2.28(m, 2H), 2.45-2.61(m, 4H), 2 .85-2.89(t, J=6Hz, 2H), 3.40-3.52(m, 2H), 3.77-3.82(m, 2H), 3.95-4.12(m , 4H), 4.21-4.32(m, 2H), 5.20-5.23(m, 1H), 7.00-7.20(m, 3H), 8.42(m, 2H); 13C NMR (CDCl3, 100MHz): δ14.05, 14.09, 19.77, 19.83, 21.23, 22.60, 22.65, 27.25, 27.33, 27.38, 27.43, 29.18, 29.22, 29.28, 29.44, 29.55, 29.58, 30.06, 31.66, 32.21, 32.26, 32.37, 33.92, 34.31, 34.33, 34 .39, 34.50, 38.40, 38.54, 45.60, 45.68, 61.97, 62.03, 62.80, 62.88, 68.66, 68.69, 70.59, 70.62, 74.68, 74.94, 123.69, 149.41, 149.80, 171.71, 171.84, 172.05, 172.09, 172.49, 175.75, 176.01, 176.22.

[0236]

change

[0237] 2-15. 3-((3-((R)-4-((3-(4-((Dimethylamino)methyl)phenyl)propanoyl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanoyl)oxy)propane-1,2-diyl bis(2-hexyldecanoate) [3-((3-((R)-4-((3-(4-((Dimethylamino)methyl)phenyl)propanoyl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanoyl)oxy]propane-1,2-diyl bis(2-hexyldecanoate)] (Compound 23)

[0238] 3-(4-((dimethylamino)methyl)phenyl)propanoic acid (compound 78), 42.38 mg, 1.05 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (59.73 mg, 1.6 equiv.) and DMAP (4.75 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature under argon gas for 20 minutes. Compound 72 (150 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C, and the mixture was added dropwise with compound 78 at 5°C for 30 minutes. The mixture was then stirred at room temperature for 13 hours. After TLC (SiO2, EtOAc / MeOH 1:1 and 1:0, PMA stain) confirmed the reaction was complete, DCM (50 mL) was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, 100% DCM, 28% aqueous NH3) to give compound 23 (127 mg, 68%) as a clear oil.

[0239] 1 H NMR (CDCl3, 400MHz): δ0.78-0.82(m, 15H), 0.91(s, 3H), 1.18-1.22(m, 41H), 1.36-1.3 7(m, 4H), 1.38-1.49(m, 4H), 2.16(s, 6H), 2.21-2.27(m, 2H), 2.44-2.57(m, 3H), 2.91(t , J=6Hz, 2H), 3.27-3.34(m, 2H), 3.43-3.56(m, 2H), 3.66(d, J=6Hz, 1H), 3.71-3.81(t, J =8Hz, 1H), 3.96-4.34(m, 5H), 5.21-5.26(m, 1H), 6.97-6.99(m, 1H), 7.06-7.19(m, 4H); 13C NMR (CDCl3, 100MHz): δ14.07, 14.11, 19.75, 19.78, 21.61, 12.20, 22.61, 22.66, 27.32, 27.34, 27.39, 27.45 , 29.20, 29.23, 29.29, 29.45, 29.59, 30.77, 31.67, 31.87, 32.23, 32.23, 32.37, 33.92, 34.37, 34.45, 35.95, 38.37, 38.50, 45.30, 45.61, 45.86, 61.98, 62.03, 62.81, 62.89, 63.98, 68.67, 70.40, 70.44, 74.44, 74.67, 128.17, 129.43, 136.60, 139.22, 171.71, 171.81, 172.12, 172.17, 173.32, 175.71, 175.99, 176.06, 176.16.

[0240] [ka]

[0241] 2-16-1.Heptadecan-9-yl(R)-8-((6-oxo-6-(undecyloxy)hexyl)(2-((3-(2,2,5,5-tetramethyl-1,3-dioxane-4-carboxamido)propanoyl)oxy)ethyl)amino)octanoate] (80)

[0242] Carboxylic acid compound 52 (30 mg, 1.5 equiv.) was dissolved in DCM (30 mL) in a reaction vessel, followed by the addition of EDCI·HCl (18 mg, 1.5 equiv.) and DMAP (2 mg, 0.2 equiv.) and vigorously stirring under argon gas at 5 °C for 10 min. Compound 79 (55 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and stirred at room temperature for 11 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 2:8), DCM (50 mL) was poured into a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (30 mL). The separated organic layer was filtered with anhydrous MgSO4 to remove water, and the filtrate was distilled under reduced pressure and purified by column chromatography (SiO2, EtOAc / hexane 8:2) to obtain compound 80 (50 mg, 68%) as a clear liquid.

[0243] 1H NMR (CDCl3, 400MHz): δ0.87(t, J=7.1Hz, 9H), 0.96(s, 3H), 1.03(s, 3H), 1.20-1.35(m, 48H), 1.35 -1.55(m, 15H), 1.55-3.73(m, 7H), 2.26(qui, J=3.9, 4H), 2.30-2.44(m, 4H), 2.54(t, J=4.7Hz, 2H ), 2.66(t, J=4.7Hz, 2H), 3.25-3.28(d, J=5.7Hz, 1H), 3.43-3.61(m, 2H), 3.66-3.69(d, J=5.1Hz, 1H), 4.02-4.11(m, 3H), 4.12-4.14(t, J=4.9Hz, 2H), 4.82-4.87(qui, J=4.7Hz, 1H), 6.95(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.07, 18.67, 18.82, 22.07, 22.63, 24.91, 25.09, 2 5.28, 25.90, 26.94, 27.16, 27.28, 28.63, 29.20, 29.22, 29.26, 29.30, 29.4 7, 29.50, 29.55, 31.83, 31.87, 32.94, 34.12, 34.19, 34.66, 52.14, 54.54, 5 4.74, 64.40, 71.46, 74.04, 77.13, 98.96, 169.66, 172.05, 173.54, 173.71.

[0244] 2-16-2. Heptadecan-9-yl(R)-8-((2-((3-(2,4-dihydroxy-3,3-dimethylbutanamido)propanoyl)oxy)ethyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (Compound 24)

[0245] Compound 80 (420 mg, 1 equiv.) and DTT (16 mg, 2 equiv.) were dissolved in DCM in a reaction vessel by stirring, followed by the addition of p-toluenesulfonic acid (4.5 mg, 0.5 equiv.) and stirring at room temperature for 2 hours. After confirming the completion of the reaction by TLC, the mixture was transferred to a separatory funnel, extracted with EtOAc (30 mL x 2), washed with saturated aqueous NaCl, and the organic layer was removed with anhydrous Na2SO4. The filtrate was then distilled under reduced pressure to remove the solvent, and purified by column chromatography (SiO2, EtOAc / hexane 1:1) to obtain compound 24 (21 mg, 44%) as a pale yellow oil.

[0246] 1 1H NMR (CDCl3, 400 MHz): δ 0.79 - 0.84 (m, 12H), 0.97 (s, 3H), 1.15 - 1.35 (m, 44H), 1.40 - 1.51 (m, 6H), 1.55 - 1.65 (m, 6H), 2.11 (qui, J = 3.3, 4H), 2.30 (m, 4H), 2.57 (m, 2H), 2.68 (t, J = 5.6 Hz, 2H), 3.27 (d, J = 4.2 Hz, 1H), 3.43 (m, 2H), 3.69 (d, J = 6.2 Hz, 1H), 4.11 - 4.12 (m, 3H), 4.12 - 4.14 (t, J = 5.0 Hz, 2H), 4.78 (qui, J = 4.7 Hz, 1H), 7.34 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.08, 18.55, 18.8, 22.04, 22.65, 24.44, 25.45, 25.23, 25.44, 26.87, 27.22, 28.64, 28.91, 29.13, 29.17, 29.23, 29.26, 29.33, 29.5, 29.53, 29.53, 29.58, 26.6, 29.7, 31.86, 32.94, 34.33, 34.19, 34.67, 52.55, 54.56, 54.78, 64.42, 71.47, 74.05, 77., 98.95, 169.55, 171.99, 173.55, 173.61.

[0247]

Chem.

[0248] 2-17. 5-((2-Hexyldecyl)oxy)-5-oxopentanoic acid (82)

[0249] 2-Hexyl-1-decanoic acid (500 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (50 mL). Glutaric anhydride (compound 81, 470 mg, 2 equiv.) and DMAP (630 mg, 2.5 equiv.) were then added and vigorously stirred at room temperature for 13 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / Hexane 1:1, PMA stain), 2N aqueous HCl (10 mL) was added to the reaction mixture and extracted with DCM (3 × 25 mL). The combined organic layers were washed with 2N aqueous HCl (2 × 20 mL) to remove residual base, followed by a saturated aqueous NaCl solution (30 mL). The separated organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9->2:8) to give compound 82 (603 mg, 82%) as a clear liquid.

[0250] 1 H NMR (CDCl3, 400MHz): δ0.79-0.82 (t, J=5.1Hz, 6H), 1.19 (m, 25H), 1.54 (br.s, 1H), 1.87-1.92(qui, J=5.2Hz, 2H), 2.31-2.38(m, 4H), 3.92(d, J=7.2Hz, 2H); 13 C NMR (CDCl3, 100MHz): δ14.07, 14.08, 19.86, 22.64, 22.67, 26.69, 29.30, 29.54, 29.60, 29.93, 31.25, 31.80, 31.89, 33.06, 33.25, 37.27, 67.39, 173.10, 179.25.

[0251] [ka]

[0252] 2-18-1.Undecyl(R)-33-(2,2,5,5-tetramethyl-1,3-dioxane-4-carboxamido)propanoate (83)

[0253] Acetal-protected pantothenic acid compound 52 (195 mg, 1.3 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (167 mg, 1.5 equiv.) and DMAP (14 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at 5 °C for 10 min under argon gas. Undecan-1-ol (100 mg, 1 equiv.) dissolved in DCM (10 mL) was then added dropwise to the reaction mixture at 5 °C and stirred at room temperature for 9 h. After confirming completion of the reaction by TLC (SiO2, 2:8 EtOAc / hexane, PMA stain), the mixture was transferred to a separatory funnel with additional DCM (50 mL) and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9->2:8) to give compound 83 (206 mg, 86%) as a clear oil.

[0254] 1 H NMR (CDCl3, 400MHz): δ0.79-0.82(t, J=5.6Hz, 3H), 0.89(s, 3H), 0.97(s, 3H), 1.19-1.23(m, 16H), 1.35(s, 3H), 1.39(s, 3H), 1.51-1.58(qui, 2H) 13 C NMR (CDCl3, 100MHz): δ14.09, 18.68, 18.81, 22.08, 22.66, 25.87, 28.59, 29.22, 29.30, 29. 46, 29.48, 29.55, 29.57, 31.88, 32.96, 34.17, 34.24, 64.89, 71.49, 98.99, 169.68, 172.19.

[0255] 2-18-2.Undecyl(R)-3-(2,4-dihydroxy-3,3-dimethylbutanamido)propanoate (84)

[0256] Compound 83 (200 mg, 1 equiv.) and DTT (149 mg, 2 equiv.) were dissolved in DCM, followed by p-toluenesulfonic acid (84 mg, 0.5 equiv.) and stirring at room temperature for 1 hour. After the reaction was completed, the mixture was extracted with EtOAc (30 mL x 2). The combined organic layers were washed with saturated aqueous NaCl. The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:1->8:2) to give compound 84 (124 mg, 69%) as a clear oil.

[0257] 1 H NMR (CDCl3, 400MHz): δ0.79-0.83(m, 6H), 0.90(s, 3H), 1.19-1.23(m, 16H), 1.51-1.58(qui, J=6.7Hz, 2H ), 2.47-2.50(t, J=5.9Hz, 2H), 3.40-3.53(m, 4H), 3.39(s, 1H), 3.99-4.03(t, J=8.7Hz, 2H), 7.31(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.08, 20.26, 21.04, 22.64, 25.87, 28.53, 29.22, 29.29, 29.48, 19.55, 29.56, 31.86, 34.03, 34.62, 39.22, 65.07, 71.09, 172.36, 173.59.

[0258] 2-18-3.(R)-3-Hydroxy-2,2-dimethyl-4-oxo-4-((3-oxo-3-(undecyloxy)propyl)amino)butyl 3-(dimethylamino)propanoate (85)

[0259] Compound 59 (42.77 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (102 mg, 1.6 equiv.) and DMAP (8 mg, 0.2 equiv.) were added and stirred at room temperature for 20 minutes under argon gas. Compound 84 (124 mg, 1 equiv.) was then dissolved in DCM (10 mL) in another reaction vessel and cooled to 5 °C. Compound 59 was added dropwise at 5 °C for 30 minutes and stirred at room temperature for 12 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / MeOH 1:1, PMA stain), the reaction mixture was evaporated under reduced pressure to remove the solvent. An additional 50 mL of DCM was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was then purified by flash column chromatography (SiO2, DCM / MeOH 9:1, 28% aqueous NH3 solution) to give compound 85 (110 mg, 70%) as a clear oil.

[0260] 1 H NMR (CDCl3, 400MHz): δ0.79-0.83(t, J=5.7Hz, 3H), 0.89(s, 3H), 0.92(s, 3H), 1.19-1.23(m, 16H), 1.35(s, 3H), 1.39(s, 3H), 1.51-1.58(qui, J=6.6H z, 2H), 2.16(s, 6H), 2.46-2.50(m, 6H), 3.19-3.22(d, J=8.2Hz, 1H), 3.37- 3.56(m, 2H), 3.60-3.62(d, J=8.7Hz, 1H), 4.00-4.04(m, 3H), 6.89(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.08, 18.69, 18.81, 22.08, 22.67, 25.87, 28.55, 29.23, 29.31, 29. 46, 29.47, 29.54, 29.58, 31.88, 32.97, 34.18, 34.24, 64.88, 71.50, 98.98, 169.67, 172.18.

[0261] 2-Hexyldecyl((R)-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazahexacosan-9-yl)glutarate (Compound 25)

[0262] Compound 82 (98 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (61 mg, 1.5 equiv.) and DMAP (5 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 85 (100 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the reaction mixture was stirred at room temperature for 14 h. After TLC (SiO2, EtOAc / hexane 7:3, PMA stain) confirmed the reaction was complete, DCM (50 mL) was added, the mixture was transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 5:5->8:2) to give compound 25 (149 mg, 87%) as a clear oil.

[0263] 1 H NMR (CDCl3, 400MHz): δ0.88-0.91(t, J=5.1Hz, 9H), 1.07-1.08(m, 6H), 1.25-1 .40(m, 42H), 1.59-1.64(m, 3H), 1.97-2.02(qui, J=6.9Hz, 2H), 2.39(s, 6H), 2. 41-2.48(t, J=6.1Hz, 2H), 2.50-2.74(m, 8H), 3.36-3.42(m, 1H), 3.56-3.62(m, 1H), 3.81-4.06(m, 4H), 4.08-4.11(t, J=8.1Hz, 2H), 4.88(s, 1H), 7.44(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.09, 14.11, 20.06, 20.27, 22.05, 22.64, 22.67, 25.91, 26.64, 26.69, 28.57, 29.31, 29.52, 29.58, 29.61, 29.95, 31.21, 31 .81, 31.90, 33.07, 33.10, 33.26, 33.82, 35.04, 37.21, 37.28, 45.22, 55. 13, 64.90, 67.34, 69.60, 76.63, 167.85, 171.72, 171.99, 172.34, 173.06.

[0264] [ka]

[0265] 2-Hexyldecyl 3-((R)-2,2,5,5-tetramethyl-1,3-dioxane-4-carboxamido)propanoate (86)

[0266] Acetal-protected pantothenic acid compound 52 (107 mg, 1.2 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (119 mg, 1.5 equiv.) and DMAP (10 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at 5 °C for 10 min under argon gas. 2-Hexyldecan-1-ol (100 mg, 0.41 mmol) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 8 h. After confirming completion of the reaction by TLC (SiO2, 3:7 EtOAc / hexane, PMA stain), an additional 50 mL of DCM was added, the mixture was transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9->4:6) to give compound 86 (177 mg, 89%) as a clear oil.

[0267] 1H NMR (CDCl3, 400MHz): δ0.87-0.90(t, J=5.2Hz, 3H), 0.97(s, 3H), 1.05(s, 3H), 1.27-1.32(m, 25H), 1.43(s, 3H), 1.47(s, 3H), 1.62(br.s, 1H), 2.55 -2.58(m, 2H), 3.27-3.30(d, J=9.2Hz, 1H), 3.45-3.63(m, 2H), 3.67-3.70 (d, J=8.1Hz, 1H), 4.00(d, J=9.8Hz, 2H), 4.08(s, 1H), 6.95-6.96(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.07, 14.09, 18.68, 18.82, 22.09, 22.63, 22.66, 26.64, 26.67, 29.29, 29.46, 29.54, 29.58, 29.92, 31.20, 31.79, 31.87, 32.69, 34.14, 34.24, 37.27, 67.56, 71.49, 77.15, 98.99, 169.69, 172.30.

[0268] 2-Hexyldecyl 3-((R)-2,4-dihydroxy-3,3-dimethylbutanamido)propanoate (87)

[0269] Compound 86 (177 mg, 1 equiv.) and DTT (122 mg, 2 equiv.) were dissolved in DCM, followed by the addition of p-toluenesulfonic acid (64 mg, 0.5 equiv.) and stirring at room temperature for 55 minutes. Upon completion of the reaction, the mixture was extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with saturated aqueous NaCl, dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:1->7:3) to give compound 87 (105 mg, 65%) as a clear oil.

[0270] 1H NMR (CDCl3, 400MHz): δ0.79-0.83(m, 9H), 0.92(s, 3H), 1.19-1.23(m, 25H), 2.47-2.51(t , J=6.2Hz, 2H), 1.55(br.s, 1H), 3.38-3.54(m, 4H), 3.91-3.94(m, 3H), 7.22-7.25(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.05, 14.03, 22.05, 22.64, 22.63, 26.64, 26.63, 29.29, 29.44, 29.55, 29.58, 29.91, 31.20, 31.79, 31.86, 32.69, 34.14, 34.24, 37.27, 67.56, 71.49, 77.15, 98.99, 169.69, 172.30.

[0271] 2-Hexyldecyl 3-((R)-4-((3-(dimethylamino)propanoyl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanoate (88)

[0272] Compound 59 (29 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (69 mg, 1.6 equiv.) and DMAP (5 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes under argon gas. Compound 87 (100 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C and the mixture containing compound 59 was added dropwise at 5°C for 30 minutes. The mixture was then stirred at room temperature for 8 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / MeOH 1:1, PMA stain), the reaction mixture was evaporated under reduced pressure to remove the solvent. Additional DCM (50 mL) was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, DCM / MeOH 9:1 / 28% aqueous NH3) to give compound 88 (84 mg, 69%) as a clear oil.

[0273] 1H NMR (CDCl3, 400MHz): δ0.79-0.82(m, 9H), 1.12(s, 3H), 1.19-1.31(m, 25H), 1.53(br.s, 1H), 2.16(s, 6H), 2.38-2. 66(m, 6H), 3.38-3.53(m, 3H), 3.81(s, 1H), 3.91(d, J=6.2Hz, 2H), 4.23-4.26(d, J=8.2Hz, 1H), 7.39-7.40(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.08, 14.10, 19.31, 22.49, 22.63, 22.66, 26.64, 26.68, 29.29, 29.55, 29.58, 29.93, 31.18, 31.8 0, 31.88, 33.33, 34.22, 34.56, 37.25, 38.48, 37.25, 38.48, 45.05, 55.75, 67.56, 71.49, 74.47, 172.17, 172.24, 172.62.

[0274] 2-19-4. (9R)-17-Hexyl-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazapentacosan-9-yl(2-hexyldecyl)glutarate (Compound 26)

[0275] Compound 82 (71 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (45 mg, 1.5 equiv.) and DMAP (19 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 88 (84 mg, 1 equiv.) in DCM (10 mL) was then added dropwise to the reaction mixture at 5 °C and stirred at room temperature for 16 h. After TLC (SiO2, EtOAc / hexane 5:5, PMA stain) confirmed the reaction was complete, DCM (50 mL) was added, and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was distilled under reduced pressure to remove the solvent, and the remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 5:5->6:4) to give compound 26 (120 mg, 88%) as a clear oil.

[0276] 11H NMR (CDCl3, 400 MHz): δ 0.79 - 0.82 (m, 12H), 0.98 - 0.99 (m, 6H), 1.11 - 1.36 (m, 47H), 1.53 (br.s, 2H), 1.88 - 1.92 (qui, J = 7.3 Hz, 2H), 2.21 (s, 6H), 2.31 (t, J = 6.7 Hz, 2H), 2.41 - 2.65 (m, 8H), 3.27 - 3.32 (m, 1H), 3.46 - 3.53 (m, 1H), 3.72 - 3.75 (d, J = 9.3 Hz, 1H), 3.38 - 3.94 (m, 5H), 4.79 (s, 1H), 7.38 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.09, 20.05, 20.27, 22.04, 22.64, 22.66, 26.64, 26.69, 23.30, 29.56, 29.60, 29.94, 31.16, 31.21, 31.81, 31.88, 33.09, 33.24, 33.78, 35.04, 37.20, 37.24, 37.27, 24.21, 55.13, 67.33, 67.57, 69.59, 76.61, 76.73, 77.05, 77.25, 167.84, 171.69, 191.97, 172.43, 173.04。

[0277]

Chem.

[0278] 2-20-1.(R)-3-Hydroxy-2,2-dimethyl-4-oxo-4-((3-oxo-3-(undecyloxy)propyl)amino)butyl 3-morpholinopropanoate (89)

[0279] Compound 75 (140 mg, 1.1 equiv.) was dissolved in DCM (20 mL) in a reaction vessel. EDCI·HCl (245 mg, 1.6 equiv.) and DMAP (20 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at room temperature for 20 minutes under argon gas. Compound 84 (300 mg, 1 equiv.) was dissolved in DCM (10 mL) in another reaction vessel and cooled to 5 °C. Compound 75 was added dropwise at 5 °C for 30 minutes and stirred at room temperature for 10 hours. After TLC (SiO2, EtOAc / MeOH 1:1, PMA stain) showed the reaction was complete, the reaction mixture was evaporated under reduced pressure to remove the solvent, and the mixture was transferred to a separatory funnel with additional DCM (50 mL) and washed with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The remaining mixture was purified by flash column chromatography (SiO2, DCM / MeOH 7:3, 28% aqueous NH3 solution) to give compound 89 (300 mg, 73%) as a clear oil.

[0280] 1 H NMR (CDCl3, 400MHz): δ0.78-0.85(t, J=5.8Hz, 3H), 0.90(s, 3H), 0.91(s, 3H), 1.20-1.25(m, 16H), 1.36(s, 3H), 1.40(s, 3H), 1.52-1.59(qui, J=6.8H z, 2H), 2.18(s, 4H), 2.47-2.55(m, 6H), 3.20-3.44(d, J=8.4Hz, 1H), 3.38- 3.60(m, 2H), 3.61-3.65(d, J=8.9Hz, 5H), 4.11-4.14(m, 3H), 6.92(s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.09, 18.70, 18.84, 22.09, 22.64, 25.89, 28.54, 29.22, 29.32, 29. 47, 29.49, 29.57, 29.59, 31.87, 32.95, 34.17, 34.23, 64.84, 71.55, 98.94, 169.63, 172.13.

[0281] 2-20-2. (R)-3,3-Dimethyl-4-((3-morpholinopropanoyl)oxy)-1-oxo-1-((3-oxo-3-(undecyloxy)propyl)amino)butan-2-yl(2-hexyldecyl)glutarate (Compound 27)

[0282] Compound 82 (270 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (167 mg, 1.5 equiv.) and DMAP (14 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 89 (300 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 11 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 4:6, PMA stain), an additional 100 mL of DCM was added and the mixture was washed with saturated aqueous NaHCO3 (2 × 100 mL) and saturated aqueous NaCl (50 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was distilled under reduced pressure to remove the solvent, and the remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 4:6->6:4) to give compound 27 (433 mg, 87%) as a clear oil.

[0283] 1 H NMR (CDCl3, 400MHz): δ0.88-0.92(t, J=5.5Hz, 9H), 1.07-1.09(m, 6H), 1.25-1.43(m, 42H), 1.59-1.67(m, 3H), 1.97-2.06(qui, J=6.5Hz, 2H), 2.30 -2.34(t, J=8.3Hz, 2H), 2.42-2.64(m, J=6.5Hz, 12H), 3.38-3.58(m, 2H), 3.38-3.65(t, J=6.6Hz, 4H), 3.85-3.97(m, 6H), 4.89(s, 1H), 7.45(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.10, 14.12, 20.07, 20.28, 22.04, 22.68, 22.65, 25.95, 26.64, 26.66, 28.54, 29.34, 29.55, 29.55, 29.62, 29.97, 31.22, 31 .84, 31.92, 33.08, 33.12, 33.27, 33.84, 35.07, 37.28, 37.29, 53.49, 54. 07, 64.95, 67.37, 69.67, 76.68, 167.88, 171.77, 171.95, 172.35, 173.03.

[0284] [ka]

[0285] 2-Hexyldecyl 3-((R)-2-hydroxy-3,3-dimethyl-4-((3-morpholinopropanoyl)oxy)butanamido)propanoate (90)

[0286] Compound 75 (237 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (414 mg, 1.6 equiv.) and DMAP (33 mg, 0.2 equiv.) were added and stirred at room temperature under argon gas for 20 minutes. Compound 87 (600 mg, 1 equiv.) was dissolved in DCM (10 mL) in another reaction vessel and cooled to 5 °C. Compound 75 was added dropwise at 5 °C under argon gas and stirred at room temperature for 7 hours. After TLC (SiO2, EtOAc / MeOH 9.5:0.5, PMA stain) confirmed the reaction was complete, the reaction mixture was evaporated under reduced pressure, and then DCM (50 mL) was added. The mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 100 mL) and saturated aqueous NaCl (100 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The remaining mixture was purified by flash column chromatography (SiO2, DCM / MeOH 9:1, 28% aqueous NH3 solution) to give compound 90 (609 mg, 77%) as a clear oil.

[0287] 1H NMR(CDCl3、400MHz):δ0.80-0.83(m、6H)、0.95(s、3H)、0.97(s、3H)、1.20-1.24(m、25H)、1.55(br.s、1H)、2.30(t、J=8.2Hz、2H)、2.33(t、J=8.2Hz、2H)、2.41-2.55(m、J=6.9Hz、6H)、3.37-3.52(m、2H)、3.36-3.63(t、J=6.5Hz、4H)、3.44-3.96(m、4H)、4.93(s、1H)、7.21(s、1H); 13 C NMR(CDCl3、100MHz):δ14.07、14.10、20.01、21.02、21.13、22.74、22.68、26.66、26.63、29.32、29.54、29.60、29.61、29.94、29.96、31.22、31.80、31.88、32.07、33.18、34.64、37.25、37.30、37.44、53.46、54.03、66.75、67.75、69.11、69.29、76.57、167.88、171.55、171.90。

[0288] 2-Hexyldecyl((2R)-1-((3-((2-hexyldecyl)oxy)-3-oxopropyl)amino)-3,3-dimethyl-4-((3-morpholinopropanoyl)oxy)-1-oxobutan-2-yl)glutarate (Compound 28)

[0289] Compound 82 (474 ​​mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (294 mg, 1.5 equiv.) and DMAP (25 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 90 (600 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 12 h. After TLC (SiO2, 5:5 EtOAc / hexane, PMA stain) confirmed the reaction was complete, DCM (100 mL) was added, the mixture was transferred to a separatory funnel, and washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the remaining mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 3:7->5:5) to give compound 28 (862 mg, 91%) as a clear oil.

[0290] 1 H NMR (CDCl3, 400MHz): δ0.79-0.82(m, 12H), 0.94(s, 3H), 0.99(s, 3H), 1.19-1.23(m, 49H), 1.54(br.s, 2H), 1.88-1.92(qui, J=7.6Hz, 2H), 2.29-2 .33(t, J=8.2Hz, 2H), 2.40-2.63(m, J=6.9Hz, 12H), 3.37-3.51(m, 2H), 3 .36-3.64(t, J=6.5Hz, 4H), 3.84-3.96(m, 6H), 4.91(s, 1H), 7.20(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.08, 14.09, 20.02, 21.02, 21.10, 22.64, 22.64, 26. 64, 26.69, 29.30, 29.55, 29.59, 29.60, 29.93, 29.95, 31.17, 31.21, 31.80, 31 .88, 32.07, 33.18, 33.60, 34.64, 37.25, 37.29, 37.44, 53.45, 54.04, 66.79, 67.39, 67.71, 69.26, 69.26, 76.56, 167.89, 171.60, 171.88, 172.79, 172.97.

[0291] [ka]

[0292] 2-22. 3-((2-Hexyldecyl)oxy)-3-oxopropanoic acid (92)

[0293] Meldrum's acid (compound 91) (500 mg, 1 equiv.) and 2-hexyl-1-decanol (841 mg, 1 equiv.) were placed in a reaction vessel, dissolved in toluene (50 mL), and heated at reflux for 5 hours. After confirming the completion of the reaction by TLC (SiO2, EtOAc / hexane 5:5), the reaction mixture was cooled to room temperature and then cooled again to 4°C. Saturated aqueous NaHCO3 solution was added to the reaction mixture, and 3M aqueous HCl was added to neutralize it. The reaction mixture was then placed in a separatory funnel and extracted with EtOAc (2 x 100 mL). Both organic layers were combined and washed with saturated aqueous NaCl (1 x 100 mL). The separated organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was distilled under reduced pressure to remove the solvent, and the remaining mixture was purified by column chromatography (SiO2, EtOAc / hexane 5:5, 1% AcOH added) to obtain compound 92 (1103 mg, 97%) as a clear oil.

[0294] 1H NMR (CDCl3, 400MHz): δ0.87-0.92(m, 6H), 1.30(m, 26H), 1.68(s, 1H), 3.44-3.47(m, 2H), 4.08-4.13(m, 2H), 10.68(br.s, 1H); 13 C NMR (CDCl3, 100MHz): δ14.09, 22.66, 22.69, 26.61, 26.66, 29.32, 29.56, 29.93, 31.10, 31.10, 31.89, 31.91, 37.22, 40.80, 68.79, 167.04, 171.69.

[0295] [ka]

[0296] 2-23-1.(9H-Fluoren-9-yl)methyl(R)-3-(2,2,5,5-tetramethyl-1,3-dioxane-4-carboxamido)propanoate (93)

[0297] Acetal-protected pantothenic acid compound 52 (476 mg, 1.2 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (438 mg, 1.5 equiv.) and DMAP (38 mg, 0.2 equiv.) were added and the mixture was stirred vigorously at 5 °C for 10 min under argon gas. 9-fluorenemethanol (300 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 1 h. After confirming completion of the reaction by TLC (SiO2, 1:9 EtOAc / hexane, PMA stain), an additional 50 mL of DCM was added, the mixture was transferred to a separatory funnel, and washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (20 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9) to give compound 93 (541 mg, 81%) as a clear oil.

[0298] 1H NMR (CDCl3, 400MHz): δ0.86(s, 3H), 0.94(s, 3H), 1.27(s, 3H), 1.31(s, 3H), 2.54-2.57(t, J=8.2 Hz, 2H), 3.14-3.17(d, J=5.1Hz, 1H), 3.38-3.52(m, 2H), 3.54-3.57(d, J=5.5Hz, 1H), 3.98(s, 1H) , 4.07-4.10(t, J=8.3Hz, 1H), 4.27-4.29(d, J=7.1Hz, 2H), 6.87-6.90(s, 1H), 7.16-7.18(t, J=5 .1Hz, 2H), 7.20-7.27(t, J=5.5Hz, 2H), 7.29-7.46(d, J=5.0Hz, 2H), 7.47-7.66(d, J=4.8Hz, 2H); 13 C NMR (CDCl3, 100MHz): δ14.23, 18.67, 18.89, 21.03, 22.14, 29.45, 32.79, 32.98, 34.21, 34.25, 46.72, 46.92, 60.36, 66.66, 71 .45, 76.87, 77.19, 77.50, 99.02, 119.78, 120.09, 120.35, 124.69, 125.01, 127.16, 127.87, 141.29, 143.64, 169.82, 171.06.

[0299] 2-23-2.(9H-Fluoren-9-yl)methyl(R)-3-(2,4-dihydroxy-3,3-dimethylbutanamido)propanoate (94)

[0300] Compound 93 (500 mg, 1 equiv.) and DTT (352 mg, 2 equiv.) were dissolved in DCM, followed by the addition of p-toluenesulfonic acid (98 mg, 0.5 equiv.) at room temperature and stirring for 15 minutes. Upon completion of the reaction, the reaction mixture was extracted with EtOAc (30 mL x 2), and the combined organic layers were washed with saturated aqueous NaCl. The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:1->7:3) to give compound 94 (277 mg, 61%) as a clear oil.

[0301] 1H NMR (CDCl3, 400MHz): δ0.90(s, 3H), 0.97(s, 3H), 2.61-2.64(t, J=8.1Hz, 2H), 3.47-3.58(m, 4H), 4.00(s, 1H), 4.18-4.21(t, J=5.1H) z, 1H), 4.40-4.42(d, 2H), 7.27-7.30(m, 3H), 7.32-7.34(t, J=5.5Hz, 2H), 7.39-7.42(d, J=5.0Hz, 2H), 7.56-7.77(d, J=4.8Hz, 2H); 13 C NMR (CDCl3, 100MHz): δ14.19, 20.34, 21.05, 21.31, 34.06, 34.63, 39.26, 46.71, 53.47, 60.47, 66.62, 71.1 5, 76.82, 77.14, 77.45, 77.48, 120.09, 124.95, 127.18, 127.89, 141.29, 143.59, 171.33, 172.17, 173.54.

[0302] 2-23-3,4-((3-((9H-Fluoren-9-yl)methoxy)-3-oxopropyl)amino)-3-hydroxy-2,2-dimethyl-4-oxobutyl(2-hexyldecyl)malonate (95)

[0303] Compound 92 (181 mg, 1.1 equiv.) was placed in a reaction vessel and dissolved in DCM (20 mL). EDCI·HCl (154 mg, 1.6 equiv.) and DMAP (13 mg, 0.2 equiv.) were added and stirred at room temperature for 20 minutes under argon gas. Compound 94 (200 mg, 1 equiv.) was placed in another reaction vessel and dissolved in DCM (10 mL). The mixture was cooled to 5°C and the compound 92 mixture was added dropwise under argon gas at 5°C for 30 minutes. The mixture was then stirred at room temperature for 2 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 2:8, PMA stain), the solvent was removed by vacuum distillation. The mixture was redissolved in DCM (50 mL), transferred to a separatory funnel, and washed sequentially with saturated aqueous NaHCO3 (2 × 100 mL) and saturated aqueous NaCl (100 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was distilled under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, DCM / hexane 1:9->1:1) to give compound 95 (256 mg, 72%) as a clear oil.

[0304] 1 H NMR(CDCl3、400MHz):δ0.78-0.81(t、J=8.5Hz、6H)、0.95-0.95(d、J=8.9Hz、6H)、1.15-1.28(m、41H)、1.54(s、1H)、2.52-2.53(t、J=5.7Hz、2H)、3.32(s、2H)、3.35-3.51(m、2H)、3.83-3.99(m、4H)、4.12-4.26(t、J=5.0Hz、1H)、4.22-4.34(m、2H)、4.81(s、1H); 13 C NMR(CDCl3、100MHz):δ14.10、20.03、20.94、21.32、22.63、22.65、26.61、26.63、26.68、29.32、29.56、29.60、29.61、29.93、29.96、31.08、31.21、31.82、31.89、33.01、33.18、33.66、34.75、37.20、37.26、37.32、41.37、46.70、66.70、67.39、68.61、70.34、76.79、77.11、77.31、77.42、120.05、125.01、125.03、127.15、129.84、141.28、143.62、143.72、166.13、166.95、167.84、171.73。

[0305] 2-23-4.1-(9H-Fluoren-9-yl)-17-hexyl-9,9-dimethyl-3,7,12,14-tetraoxo-2,11,15-trioxa-6-azapentacosan-8-yl(2-hexyldecyl)glutarate (96)

[0306] Compound 82 (164 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (101 mg, 1.5 equiv.) and DMAP (7 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C. Compound 95 (250 mg, 1 equiv.) in DCM (10 mL) was then added dropwise to the reaction mixture at 5 °C and stirred at room temperature for 4 h. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 1:9, PMA stain), an additional 100 mL of DCM was added and the mixture was transferred to a separatory funnel and washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9) to give compound 96 (300 mg, 81%) as a clear oil.

[0307] 1 H NMR (CDCl3, 400MHz): δ0.78-0.81(t, J=8.5Hz, 12H), 0.95-0.97(d, J=8.9Hz, 6H), 1 .15-1.27(m, 53H), 1.53(s, 2H), (qui, J=5.1Hz, 2H), 2.26-2.30(t, J=5.5Hz, 2H), 2. 37-2.40(t, J=5.1Hz, 2H), 2.53-2.56(t, J=5.7Hz, 2H), 3.32(s, 2H), 3.35-3.52(m, 2 H), 3.83-3.99(m, 6H), 4.10-4.26(t, J=5.0Hz, 1H), 4.29-4.34(m, 2H), 4.82(s, 1H); 13C NMR (CDCl3, 100MHz): δ14.09, 14.11, 20.01, 20.92, 21.30, 22.65, 22.67, 26.60, 26.64, 26.69, 29.31 , 29.56, 29.59, 29.61, 29.93, 29.95, 31.07, 31.21, 31.81, 31.89, 33.02, 33.18, 33.67, 34.75, 37.20, 37.28, 37.32, 41.36, 46.70, 66.70, 67.35, 68.61, 70.34, 76.79, 77.11, 77.31, 77.42, 120.05, 125.01, 125.03, 127.15, 129.84, 141.28, 143.62, 143.70, 166.19, 166.98, 167.85, 171.72, 172.30, 173.01.

[0308] 2-23-5.3-(4-((3-((2-Hexyldecyl)oxy)-3-oxopropanoyl)oxy)-2-((5-((2-hexyldecyl)oxy)-5-oxopentanoyl)oxy)-3,3-dimethylbutanamido)propanoic acid (97)

[0309] A 20% piperidine solution was prepared by mixing piperidine and DMF in a 2:8 volume ratio in a reaction vessel and cooling to 0 °C. Compound 96 (300 mg) was dissolved in DMF (20 mL) in another reaction vessel and stirred at 0 °C for 5 minutes. The prepared 20% piperidine solution (40 mL) was then added under argon and stirred at 0 °C for 5 minutes. After the reaction was completed, the pH was adjusted to 7 with 3M aqueous HCl, diluted with EtOAc (200 mL), transferred to a separatory funnel, and washed sequentially with distilled water (2 × 200 mL) and saturated aqueous NaCl (2 × 200 mL). The separated organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 0:1->4:6, with 1% AcOH) to give compound 97 (226 mg, 91%) as a clear oil.

[0310] 1H NMR (CDCl3, 400MHz): δ0.82(t, J=8.1Hz, 12H), 0.97-1.00(d, J=8.3Hz, 6H), 1.15-1.29(m, 50H), 1.56(s, 2H), 1.85-1.93 (m, 2H), 2.30-2.55(m, 6H), 3.28-3.38(m, 3H), 3.55-3.63(m, 1H), 3.85-4.01(m, 6H), 4.87(s, 1H), 6.76(t, J=5.7Hz, 1H); 13 C NMR (CDCl3, 100MHz): δ14.08, 14.09, 19.86, 20.99, 21.24, 22.64, 22.66, 26.60, 26.64, 26.68, 29.30, 29.35, 29.55, 29.59, 29.94, 31.07, 31.20, 31 .80, 31.89, 32.87, 33.18, 33.39, 34.49, 37.21, 37.22, 37.25, 41.32, 67.7 9, 68.67, 70.34, 77.23, 166.30, 167.09, 167.97, 171.70, 173.96, 175.31.

[0311] 20-Hexyl-2,12,12-trimethyl-6,10,15,17-tetraoxo-14,18-dioxa-2,5,9-triazaoctacosan-11-yl(2-hexyldecyl)glutarate (Compound 29)

[0312] Compound 97 (100 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (50 mL). HATU (66 mg, 1.5 equiv.), Hunig's base (22.32 mg, 1.5 equiv.), and compound 98 (12 mg, 1.1 equiv.) were added and vigorously stirred at room temperature for 11 hours. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1, PMA stain), an additional 100 mL of DCM was added. The mixture was then transferred to a separatory funnel and washed sequentially with saturated aqueous NaHCO3 (2 x 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 10:0->7:3) to give compound 29 (89 mg, 82%) as a clear oil.

[0313] 1 1H NMR (CDCl3, 400 MHz): δ 0.81 (t, J = 7.1 Hz, 12H), 0.97 - 0.99 (d, J = 8.1 Hz, 6H), 1.14 - 1.30 (m, 50H), 1.55 (s, 2H), 1.87 - 1.93 (qui, J = 8.2 Hz, 2H), 2.14 (s, 6H), 2.17 - 2.45 (m, 8H), 3.15 - 3.51 (m, 6H), 3.87 - 4.02 (m, 6H), 4.78 (s, 1H), 6.24 (m, 1H), 7.16 - 7.22 (m, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.07, 19.99,21.03, 21.06, 22.62, 22.64, 26.58, 26.62, 26.67, 29.58, 26.62, 26.67, 29.28, 29.53, 29.57, 29.58, 29.91, 31.05, 31.20, 31.78, 31.86, 33.06, 33.21, 35.26, 35.56, 26.67, 37.19, 37.27, 41.37, 45.08, 57.67, 67.34, 68.50, 70.28, 166.27, 166.81, 167.80, 171.54, 171.99, 173.00。

[0314] [Chemical formula]

[0315] 2-24. 18-Hexyl-10,10-dimethyl-1-morpholino-4,8,13,15-tetraoxo-12,16-dioxa-3,7-diazahexacosan-9-yl(2-hexyldecyl)glutarate (Compound 30) [[ID=Compound 97 (100 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (50 mL). HATU (66 mg, 1.5 equiv.), Hunig's base (22.32 mg, 1.5 equiv.), and compound 99 (17 mg, 1.1 equiv.) were added and vigorously stirred at room temperature for 9 hours. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1, PMA stain), DCM (100 mL) was added and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 10:0 → 9:1) to give compound 30 (99 mg, 88%) as a clear oil.

[0317] 1 H NMR (CDCl3, 400MHz): δ0.81(t, J=7.9Hz, 12H), 0.97-0.99(d, J=8.2Hz, 6H), 1.15-1.31(m, 51H), 1.55(s, 2H), 1.85-1.93(qui, J=8.1Hz, 2H), 2.29- 2.44(m, 10H), 3.22-3.30(m, 2H), 3.32(s, 2H), 3.35-3.51(m, 2H), 3.63-3 .65(t, J=4.1Hz, 4H), 3.87-4.01(m, 6H), 6.15(s, 1H), 7.02-7.05(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.08, 19.99, 20.99, 21.20, 22.63, 22.65, 26.58, 26.62, 26 .67, 29.29, 29.54, 29.57, 29.59, 29.92, 29.93, 31.05, 31.19, 31.79, 31.79, 31.87 , 33.05, 33.21, 35.17, 35.42, 35.51, 37.18, 37.23, 37.26, 41.36, 53.32, 57.02, 66.78, 67.35, 66.53, 70.28, 76.95, 166.26, 166.87, 167.86, 171.32, 171.87, 172.99.

[0318] [ka]

[0319] 2-25. 20-Hexyl-2,12,12-trimethyl-6,10,15,17-tetraoxo-5,14,18-trioxa-2,9-diazaoctacosan-11-yl(2-hexyldecyl)glutarate (Compound 31)

[0320] Compound 97 (100 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (33 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 100 (11 mg, 1.1 equiv.) in DCM (10 mL) was then added dropwise to the reaction mixture at 5 °C and the mixture was stirred at room temperature for 4 h. After completion of the reaction was confirmed by TLC (SiO2, 100% EtOAc, PMA stain), additional DCM (100 mL) was added, the mixture was transferred to a separatory funnel, and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 20:0->19:1) to give compound 31 (97 mg, 90%) as a clear oil.

[0321] 1 H NMR (CDCl3, 400MHz): δ0.81 (t, J=7.5Hz, 12H), 0.97-0.99 (d, J=8.2Hz, 6H), 1.13-1.31(m, 50H), 1.56(s, 2H), 1.88-1.93(qui, J=8.2Hz, 2H), 2.22(s, 6H) , 2.31(t, J=7.6Hz, 2H), 2.41(t, J=7.1Hz, 2H), 2.47-2.52(m, 4H), 3.34(s, 2H) ), 3.37-3.52(m, 2H), 3.86-4.14(m, 8H), 4.83(s, 1H), 6.83(t, J=7.8Hz, 1H); 13C NMR (CDCl3, 100MHz): δ14.07, 14.08, 20.01, 20.98, 21.17, 22.63, 22.65, 26. 59, 26.64, 26.68, 29.29, 29.55, 29.58, 29.60, 29.93, 29.94, 31.07, 31.21, 31 .79, 31.88, 33.03, 33.20, 33.76, 34.75, 37.20, 37.29, 41.37, 45.58, 57.68, 62.14, 67.37, 68.58, 70.32, 166.21, 166.91, 167.72, 171.72, 172.36, 173.02.

[0322] [ka]

[0323] 2-26. 18-Hexyl-10,10-dimethyl-1-morpholino-4,8,13,15-tetraoxo-3,12,16-trioxa-7-azahexacosan-9-yl(2-hexyldecyl)glutarate (Compound 32)

[0324] Compound 97 (100 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (33 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 min. Compound 101 (17 mg, 1.1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 6 h. After completion of the reaction was confirmed by TLC (SiO2, 100% EtOAc, PMA stain), an additional 100 mL of DCM was added and the mixture was transferred to a separatory funnel and washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 8:3->10:0) to give compound 32 (89 mg, 79%) as a clear oil.

[0325] 11H NMR (CDCl3, 400 MHz): δ 0.82 (t, J = 7.1 Hz, 12H), 0.97 - 0.99 (d, J = 8.9 Hz, 6H), 1.13 - 1.31 (m, 51H), 1.56 (s, 2H), 1.88 - 1.93 (qui, J = 8.1 Hz, 2H), 2.23 - 2.33 (t, J = 7.5 Hz, 2H), 2.39 - 2.49 (m, 8H), 2.53 - 2.56 (t, J = 7.0 Hz, 2H), 3.34 (s, 2H), 3.37 - 3.52 (m, 2H), 3.62 - 3.64 (t, J = 3.9 Hz, 4H), 3.86 - 4.00 (m, 6H), 4.11 - 4.17 (m, 2H), 4.83 (s, 1H), 6.75 (t, J = 7.9 Hz, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.06, 19.99, 20.89, 21.28, 22.61, 22.63, 26.57, 26.62, 26.66, 29.27, 26.62, 2,6.66, 29.27, 29.52, 29.55, 29.57, 29.90, 29.92, 31.05, 31.19, 31.77, 31.86, 33.00, 33.17, 33.65, 34.73, 37.18, 37.28, 41.34, 53.78, 56.98, 61.99, 66.80, 67.69, 66.80, 67.34, 68.58, 70.30, 166.16, 166.94, 167.75, 171.68, 172.24, 172.99。

[0326]

Chem.

[0327] 2-27-1. O,O'-(4-((3-((9H-Fluoren-9-yl)methoxy)-3-oxopropyl)amino)-2,2-dimethyl-4-oxobutane-1,3-diyl)bis(2-hexyldecyl)diglutarate] (102)

[0328] Compound 82 (164 mg, 2.2 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (101 mg, 1.5 equiv.) and DMAP (7 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 minutes. Compound 94 (250 mg, 1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 4 hours. After confirming completion of the reaction by TLC (SiO2, EtOAc / hexane 1:9, PMA stain), an additional 100 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 1:9) to give compound 102 (300 mg, 81%) as a clear oil.

[0329] 1 H NMR (CDCl3, 400MHz): δ0.80(t, J=8.0Hz, 12H), 0.92-0.97(d, J=9.2Hz, 6H), 1.11-1.28(m, 50H), 1.52(s, 2H), 1.63(s, 1H), (qui, J=5.5Hz, 4H), 2.26-2.33(m, 6H), 2.38(t, J=5.7Hz, 2H), 2.53-2.56(t, J=5.9Hz, 2 H), 3.38-3.51(m, 2H), 3.76-3.97(m, 6H), 4.12-4.31(t, J=5.0Hz, 1H), 4.33(d, J=9.1Hz, 2H), 4.88(s, 1H) , 6.61(m, 1H), 7.24(t, J=7.1Hz, 2H), 7.26(t, J=8.1Hz, 2H), 7.49(d, J=8.3Hz, 2H), 7.68(d, J=8.2Hz, 2H); 13C NMR (CDCl3, 100MHz): δ14.08, 14.10, 20.00, 20.13, 20.90, 21.32, 22.64, 22.67, 26.65, 26 .69, 29.30, 29.56, 29.60, 29.95, 31.21, 31.24, 31.81, 31.89, 33.05, 33.17, 33.24, 33.37, 33.66, 34.65, 37.29, 37.38, 46.70, 66.71, 67.35, 67.41, 69.23, 120.08, 124.98, 125.00, 127.17, 127.88, 141.30, 143.57, 143.65, 167.95, 171.62, 172.48, 172.62, 173.01, 173.09.

[0330] 2-27-2,3-(2,4-bis((5-((2-Hexyldecyl)oxy)-5-oxopentanoyl)oxy)-3,3-dimethylbutanamido)propanoic acid [3-(2,4-bis((5-((2-Hexyldecyl)oxy)-5-oxopentanoyl)oxy)-3,3-dimethylbutanamido)propanoic acid] (103)

[0331] A 20% piperidine solution was prepared by mixing piperidine and DMF in a volume ratio of 2:8 in a reaction vessel and stirring the mixture. The mixture was then cooled to 0°C. Compound 102 (300 mg) was dissolved in DMF (20 mL) in another reaction vessel, stirred for 5 minutes, and cooled to 0°C. A previously prepared 20% piperidine solution (40 mL) was added under argon gas and stirred at 0°C for 5 minutes. Upon completion of the reaction, 3M aqueous HCl was added to the reaction mixture to adjust the pH to 7. EtOAc (200 mL) was added, and the mixture was transferred to a separatory funnel and washed sequentially with distilled water (2 x 200 mL) and saturated aqueous NaCl (2 x 200 mL). The separated organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and the resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 0:1->4:6, 1% AcOH added) to give compound 103 (226 mg, 91%) as a clear oil.

[0332] 1H NMR (CDCl3, 400MHz): δ0.81(t, J=8.1Hz, 12H), 0.91-0.92(d, J=9.0Hz, 6H), 1.12-1.21(m, 51H), 1.52(s, 2H), 1.62(s, 1H), (qui, J=5.5Hz, 4 H), 2.27-2.33(m, 6H), 2.39(t, J=5.8Hz, 2H), 2.53-2.55(t, J=5.3Hz, 2H), 3.38-3.52(m, 2H), 3.77-3.97(m, 6H), 4.89(s, 1H), 6.62(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.09, 14.11, 20.02, 20.14, 20.91, 21.31, 22.66, 22.68, 26.61, 26.65, 29.31, 29.56, 29.61, 29.95, 31.22, 31.24, 31.81, 31 .89, 33.05, 33.17, 33.24, 33.37, 33.66, 34.65, 37.29, 37.38, 46.70, 66.7 1, 67.35, 67.41, 69.23, 167.95, 171.62, 172.48, 172.62, 173.01, 173.09.

[0333] 2-27-3. O,O'-(4-((3-((2-(Dimethylamino)ethyl)amino)-3-oxopropyl)amino)-2,2-dimethyl-4-oxobutane-1,3-diyl)bis(2-hexyldecyl)diglutarate] (Compound 33)

[0334] Compound 103 (100 mg, 1 equiv.) was dissolved in DCM (50 mL) in a reaction vessel. HATU (66 mg, 1.5 equiv.), Hunig's base (22.32 mg, 1.5 equiv.), and compound 98 (12 mg, 1.1 equiv.) were then added and vigorously stirred at room temperature for 11 hours. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1, PMA stain), DCM (100 mL) was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 x 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 10:0->7:3) to give compound 33 (89 mg, 82%) as a clear liquid.

[0335] 1 1H NMR (CDCl3, 400 MHz): δ 0.81 (t, J = 7.1 Hz, 12H), 0.97 - 0.99 (d, J = 8.1 Hz, 6H), 1.14 - 1.30 (m, 50H), 1.55 (s, 2H), 1.87 - 1.93 (qui, J = 8.2 Hz, 4H), 2.14 (s, 6H), 2.17 - 2.45 (m, 8H), 3.38 - 3.51 (m, 2H), 3.37 (d, J = 9.6 Hz, 2H), 3.91 (d, J = 9.7 Hz, 4H), 3.96 (d, J = 9.9 Hz, 2H), 4.10 - 4.14 (m, 2H), 4.87 (s, 1H), 6.24 (m, 1H), 7.16 - 7.22 (m, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.07, 19.99, 21.03, 21.06, 22.62, 22.64, 26.58, 26.62, 26.67, 29.58, 26.62, 26.67, 29.28, 29.53, 29.57, 29.58, 29.91, 31.05, 31.20, 31.78, 31.86, 33.06, 33.21, 35.26, 35.56, 26.67, 37.19, 37.27, 41.37, 45.08, 57.67, 67.34, 68.50, 70.28, 166.27, 166.81, 167.80, 171.54, 171.99, 173.00.

[0336]

Chem.

[0337] 2-28. O,O'-(2,2-Dimethyl-4-((3-((2-morpholinoethyl)amino)-3-oxopropyl)amino)-4-oxobutane-1,3-diyl)bis(2-hexyldecyl)diglutarate (Compound 34)

[0338] Compound 103 (100 mg, 1 equiv.) was dissolved in DCM (50 mL) in a reaction vessel. HATU (66 mg, 1.5 equiv.), Hunig's base (22.32 mg, 1.5 equiv.), and compound 99 (17 mg, 1.1 equiv.) were added and vigorously stirred at room temperature for 9 hours. After completion of the reaction was confirmed by TLC (SiO2, EtOAc / MeOH 9:1, PMA stain), an additional 100 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 x 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over MgSO4, filtered, and the solution was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / MeOH 10:0->9:1) to give compound 34 (99 mg, 88%) as a clear oil.

[0339] 1 H NMR (CDCl3, 400MHz): δ0.81-0.82(t, J=7.0Hz, 12H), 0.95-0.98(d, J=8.5Hz, 6H), 1 .11-1.35(m, 50H), 1.54(s, 2H), 1.83-1.92(m, 4H), 2.28-2.47(m, 16H), 3.21-2.32 (m, 4H), 3.40(t, J=3.2Hz, 4H), 3.78-3.81(d, J=8.2Hz, 1H), 3.89-3.91(d, J=8.0Hz , 4H), 3.94-3.97(d, J=9.2Hz, 1H), 4.84(s, 1H), 6.18(m, 1H), 7.00(t, J=6.9Hz, 1H); 13 C NMR (CDCl3, 100MHz): δ14.04, 14.05, 19.97, 20.09, 20.90, 21.30, 22.59, 22.62, 26.61, 26.65, 29.25, 29.51, 29.56, 29.90, 31.19, 31.31.76, 31.84, 33.05, 33.1 8, 33.32, 35.07, 35.37, 35.54, 37.25, 53.33, 57.06, 66.76, 67.28, 67.34, 69.21, 76.77, 76.91, 77.09, 77.41, 167.99, 171.45, 191.77, 172.57, 172.95, 173.02.

[0340] [ka]

[0341] 2-29. O,O'-(4-((3-((2-(Dimethylamino)ethyl)amino)-3-oxopropyl)amino)-2,2-dimethyl-4-oxobutane-1,3-diyl)bis(2-hexyldecyl)diglutarate] (Compound 35)

[0342] Compound 103 (100 mg, 1 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (33 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5 °C for 10 minutes. Compound 100 (11 mg, 1.1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the mixture was stirred at room temperature for 4 hours. After confirming completion of the reaction by TLC (SiO2, 100% EtOAc, PMA stain), additional DCM (100 mL) was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure and purified by column chromatography (SiO2, EtOAc / MeOH 20:0->19:1) to give compound 35 (97 mg, 90%) as a clear oil.

[0343] 1 H NMR (CDCl3, 400MHz): δ0.81-0.82(t, J=7.1Hz, 12H), 0.95-0.98(d, J=8.1Hz, 6H), 1.11-1.32 (m, 50H), 1.54(s, 2H), 1.85-1.91(qui, J=8.0Hz, 4H), 2.22(s, 6H), 2.30-2.32(t, J=7.0Hz, 6H ), 2.39(t, J=7.9Hz, 2H), 2.43-2.51(m, 4H), 3.37(d, J=9.6Hz, 2H), 3.38-3.51(m, 2H), 3.91(d , J=9.7Hz, 4H), 3.96(d, J=9.9Hz, 2H), 4.10-4.14(m, 2H), 4.87(s, 1H), 6.77(t, J=6.9Hz, 1H); 13C NMR (CDCl3, 100MHz): δ14.04, 19.99, 20.09, 20.79, 21.32, 22.60.22.62, 26.61, 26.66, 29.26, 29.52, 29.56, 29.91, 31.20, 31.77, 31.85, 33.02 , 33.16, 33.19, 33.33, 33.75, 34.66, 37.26, 37.30, 45.57, 57.68, 62.16 , 67.28, 67.34, 69.21, 167.80, 171.59, 172.44, 192.55, 172.95, 173.01.

[0344] [ka]

[0345] 2-30. O,O'-(2,2-Dimethyl-4-((3-((2-morpholinoethyl)amino)-3-oxopropyl)amino)-4-oxobutane-1,3-diyl)bis(2-hexyldecyl)diglutarate (Compound 36)

[0346] Compound 103 (100 mg, 1.3 equiv.) was dissolved in DCM (30 mL) in a reaction vessel. EDCI·HCl (33 mg, 1.5 equiv.) and DMAP (3 mg, 0.2 equiv.) were then added and vigorously stirred under argon gas at 5 °C for 10 minutes. Compound 101 (17 mg, 1.1 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and the reaction mixture was stirred at room temperature for 6 hours. After confirming completion of the reaction by TLC (SiO2, 100% EtOAc, PMA stain), additional DCM (100 mL) was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (1 × 200 mL). The separated organic layer was dried over anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by flash column chromatography (SiO2, EtOAc / hexane 8:3->10:0) to give compound 36 (89 mg, 79%) as a clear oil.

[0347] 11H NMR (CDCl3, 400 MHz): δ 0.81 (t, J = 7.6 Hz, 12H), 0.94 - 0.98 (d, J = 8.7 Hz, 6H), 1.10 - 1.31 (m, 47H), 1.54 (s, 2H), 1.85 - 1.93 (m, 4H), 2.28 - 2.57 (m, 16H), 2.33 - 2.50 (m, 2H), 3.65 (m, 4H), 3.76 - 3.79 (d, 1H), 3.89 - 3.98 (m, 5H), 4.13 - 1.19 (m, 2H), 4.88 (s, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.06, 20.01, 20.12, 20.89, 21.31, 22.62, 22.64, 26.64, 26.68, 29.28, 29.54, 29.58, 29.93, 31.22, 31.79, 31.87, 33.06, 33.18, 33.23, 33.35, 33.66, 34.68, 37.29, 37.35, 53.76, 56.97, 61.62, 66.72, 67.34, 37.41, 69.21, 167.88, 171.62, 172.38, 172.59, 173.00, 173.05。

[0348]

Chem.

[0349] 2-31-1,1-((tert-butyldiphenylsilyl)oxy)octan-2-ol [1-((tert-Butyldiphenylsilyl)oxy)octan-2-ol] (105)

[0350] 1,2-octanediol (1,2-octanediol, (compound 104), 500 mg, 1 equiv.) and imidazole (279.32, 1.2 equiv.) were placed in a reaction vessel and dissolved in DCM (30 mL). After stirring for 5 minutes, a solution of tert-butyldiphenylsilyl chloride (TBDPS-Cl; 936.38 mg, 1 equiv.) in DCM (20 mL) was added dropwise over 20 minutes. The reaction mixture was vigorously stirred at room temperature for 5 hours. After confirming the completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 19:1), the reaction mixture was concentrated to a volume of 35 mL. Next, 70 mL of n-hexane was added, and the precipitated solid was filtered. The remaining filtrate was concentrated and purified by column chromatography (SiO; hexane / ethyl acetate, 39:1 to 19:1, v / v) to obtain compound 105 (1183 mg, 90%) as a transparent liquid.

[0351] 1 H NMR (CDCl3, 400MHz): δ0.98-0.95(t, J=6.2Hz, 3H), 1.14(s, 9H), 1.32(s, 7H), 1.44-1.46(m, 3H), 2 .57(s, 1H), 3.54-3.58(m, 1H), 3.72-3.79(m, 2H), 7.43-7.49(m, 6H), 7.73-7.75(d, J=6.7Hz, 4H); 13 C NMR (CDCl3, 100MHz): δ14.15, 19.10, 22.65, 25.54, 26.92, 29.39, 31.82, 32. 85, 68.12, 72.02, 127.83, 129.86, 129.86, 133.25, 133.29, 235.59, 135.61.

[0352] 2-31-2,1-((tert-Butyldiphenylsilyl)oxy)octan-2-yl octanoate [1-((tert-Butyldiphenylsilyl)oxy)octan-2-yl octanoate] (106)

[0353] Compound 104 (100 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (50 mL). Octanoic acid (62 mg, 1.3 equiv.), EDCI·HCl (46 mg, 1.5 equiv.), and DMAP (4 mg, 0.2 equiv.) were added and the mixture was heated to reflux for 18 hours. After completion of the reaction was confirmed by TLC (SiO2; hexane / ethyl acetate, 39:1), DCM (50 mL) was added and the mixture was washed sequentially with distilled water (2 × 50 mL), saturated aqueous NaHCO3 (2 × 50 mL), and brine (1 × 20 mL). The organic layer was collected and dehydrated with anhydrous MgSO4. The filtrate was then distilled under reduced pressure and purified by column chromatography (SiO2; hexane / ethyl acetate, 39:1 to 19:1, v / v) to give compound 106 (118 mg, 89%) as a clear liquid.

[0354] 1 H NMR (CDCl3, 400MHz): δ0.97-0.98(t, J=6.1Hz, 6H), 1.14(s, 9H), 1.22-1.33(m, 16H), 1.47-1.55(m, 4H), 2.17-2.22(m, 2H), 3.55-3.63(m, 2H), 4.90-4.95(m, 1H), 7.27-7.36(m, 6H), 7.57-7.59(d, J=6.8Hz, 4H); 13 C NMR (CDCl3, 100MHz): δ14.09, 19.26, 22.58, 22.63, 25.10, 25.16, 26.76, 29.00, 29.19, 30.56, 31. 69, 31.72, 34.64, 65.09, 74.20, 127.67, 129.67, 129.66, 129.69, 133.53, 135.58, 135.65, 173.51.

[0355] 2-31-3.1-Hydroxyoctan-2-yl octanoic acid [1-Hydroxyoctan-2-yl octanoate] (107)

[0356] The reaction vessel was flame-dried to remove moisture, then cooled to 20 °C and backfilled with nitrogen. Compound 106 (200 mg, 1 equiv.) and dry THF (15 mL) were then added via syringe and stirred for 1 minute. A 1 M n-Bu4NF solution (0.5 mL, 1.2 equiv.) was then added dropwise via syringe over 10 minutes, and the reaction mixture was stirred for 2 hours at 20 °C. After confirming the completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 9:1), the mixture was diluted with distilled water (2 × 50 mL) and extracted with DCM (3 × 40 mL). The organic layers were collected and washed with saturated aqueous NaCl, then dehydrated with anhydrous Na2SO4. The filtrate was filtered and evaporated under reduced pressure. The resulting mixture was purified by column chromatography (SiO2; hexane / ethyl acetate, 1:19, v / v) to give compound 107 (96 mg, 90%) as a colorless oil.

[0357] 1 H NMR (CDCl3, 400MHz): δ0.81(t, J=6.3Hz, 6H), 1.20(m, 16H), 1.47-1.55(m, 4H), 2.18-2.22(m, 2H), 3.56-3.64(m, 2H), 4.91-4.94(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.09, 19.27, 22.54, 22.65, 25.14, 25.17, 26.88, 29.54, 30.56, 31.74, 31.72, 34.77, 65.09, 74.21, 173.56.

[0358] 2-31-4,6-((2-(octanoyloxy)octyl)oxy)-6-oxohexanoic acid [6-((2-(Octanoyloxy)octyl)oxy)-6-oxohexanoic acid] (108)

[0359] Compound 107 was placed in a reaction vessel and dissolved in DCM (25 mL). Compound 81 (84 mg, 2 equiv.) and DMAP (112 mg, 2.5 equiv.) were then added and vigorously stirred at room temperature for 10 hours. After confirming completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 9:1), the mixture was acidified with 1N aqueous HCl and extracted with DCM (3 x 25 mL). The combined organic layer was washed with saturated aqueous NaCl (1 x 30 mL), removed with anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure. The mixture was purified by column chromatography (SiO2; hexane / ethyl acetate, 9:1 to 8:2, v / v) to give compound 108 (99 mg, 70%) as a clear liquid.

[0360] 1 H NMR (CDCl3, 400MHz): δ0.87(t, J=6.9Hz, 6H), 1.24-1.28(m, 18H), 1.55-1.60(m, 4H), 1.90-1.98(m, 2H), 2.27-2. 30(t, J=6.1Hz, 2H), 2.37-2.44(m, 4H), 3.98-4.02(m, 1H), 4.21-4.25(dd, J=2.2, 9.1Hz, 1H), 5.05-5.10(m, 1H); 13 C NMR (CDCl3, 100MHz): δ14.12, 19.84, 22.48, 22.55, 24.84, 25.02, 28.87, 28.98, 29.01, 30.66, 3 1.56, 31.62, 32.90, 33.25, 32.90, 33.25, 34.05, 60.41, 64.81, 71.73, 172.37, 173.60, 178.67.

[0361] 2-31-5,17-Hexyl-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazapentacosan-9-yl(2-(octanoyloxy)octyl)glutarate (Compound 37)

[0362] Compound 108 (170.90 mg, 1.2 equiv.) was dissolved in DCM (30 mL) and EDCI·HCl (113 mg, 1.6 equiv.) and DMAP (9 mg, 0.2 equiv.) were added to a reaction vessel. The mixture was stirred at 5°C for 10 minutes under argon gas. Compound 88 (200 mg, 1 equiv.) was dissolved in DCM (10 mL) and added dropwise at 5°C. The mixture was stirred at room temperature for 20 hours. After confirming the completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 7:3), an additional 50 mL of DCM was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 50 mL) and saturated aqueous NaCl (1 × 20 mL). The organic layer was filtered with anhydrous MgSO4 to remove water, and the filtrate was distilled under reduced pressure. It was then purified by column chromatography (SiO2; ethyl acetate / hexane, 2:8 to 5:5, v / v) to obtain compound 37 (292 mg, 87%) as a clear liquid.

[0363] 1 H NMR (CDCl3, 400MHz): δ0.87(t, J=6.5Hz, 12H), 0.98-0.99(d, J=4.7Hz, 6H) , 1.19(m, 47H), 1.53(m, 1H), 1.88-1.92(m, 2H), 2.21(s, 6H), 2.30-2.65(m, 12H), 3.27-3.32(m, 1H), 3.46-3.53(m, 1H), 3.72-3.75(d, J=11.2Hz, 1H), 3.86-3.94(m, 5H), 4.79(s, 1H), 5.16(bs, 1H), 7.35-7.38(t, J=5.8Hz, 1H); 13 C NMR (CDCl3, 100MHz): δ14.09, 20.05, 20.27, 22.04, 22.64, 22.66, 26.64, 26.69, 29.30, 29.56, 29.60, 29.94, 31.16, 31.21, 31.81, 31.88, 33.09 , 33.24, 33.78, 35.04, 37.20, 37.24, 37.27, 45.21, 55.13, 67.33, 67.57 , 69.59, 74.47, 76.61, 167.84, 171.69, 171.97, 172.43, 173.04, 176.04.

[0364] [ka]

[0365] 2-32-1,1-((tert-Butyldiphenylsilyl)oxy)octan-2-yl 5-(1,2-dithiolan-3-yl)pentanoate (109)

[0366] Compound 105 (500 mg, 1 equiv.) was dissolved in DCM (50 mL) in a reaction vessel. Lipoic acid (349 mg, 1.3 equiv.), EDCI·HCl (374 mg, 1.5 equiv.), and DMAP (31 mg, 0.2 equiv.) were added and the mixture was heated to reflux for 20 h. After completion of the reaction was confirmed by TLC (SiO2; hexane / ethyl acetate, 29:1), additional DCM (50 mL) was added, and the organic layer was washed sequentially with distilled water (2 × 100 mL), saturated aqueous NaHCO3 (2 × 100 mL), and saturated aqueous NaCl (1 × 40 mL). The resulting organic layer was removed with anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by column chromatography (SiO2; hexane / ethyl acetate, 49:1 to 39:1, v / v) to give compound 109 (603 mg, 81%) as a yellow liquid.

[0367] 1 H NMR (CDCl3, 400MHz): δ0.85-0.88(t, J=6.1Hz, 3H), 1.04(s, 9H), 1.25(s, 8H), 1.44-1.66(m, 8H), 1.68-1.89(s, 1H), 2.26-2.31(m, 2 H), 2.42-2.43(m, 1H), 3.09-3.16(m, 2H), 3.51-3.55(m, 1H), 3.66-3.68(m, 2H), 5.00(m, 1H), 7.25-7.42(m, 6H), 7.65-7.66(m, 4H); 13C NMR (CDCl3, 100MHz): δ14.10, 19.27, 22.59, 24.79, 25.17, 26.78, 28.82, 29.18, 30.55, 31.71, 34.33, 34.33, 34.66, 38.49, 40.21, 56.12. 65.09, 74.38, 127.69, 129.69, 129.73, 133.45, 133.52, 135.57, 135.64, 173.10.

[0368] 2-32-2,1-Hydroxyoctan-2-yl 5-(1,2-dithiolan-3-yl)pentanoate (110)

[0369] The reaction vessel was dried over flame, cooled to 20 °C, and backfilled with nitrogen. Compound 109 (600 mg, 1 equiv.) and dry THF (30 mL) were then added via syringe and stirred for 1 minute. A 1 M n-Bu4NF solution (1.26 mL, 1.2 equiv.) was then added dropwise via syringe over 10 minutes, and the reaction mixture was stirred at 20 °C for 1 hour. After confirming completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 8:2), the mixture was diluted with distilled water (2 × 50 mL) and extracted with DCM (3 × 40 mL). The combined organic layer was washed with saturated aqueous NaCl, then dried over anhydrous Na2SO4. The filtrate was filtered, and the mixture was evaporated under reduced pressure. The mixture was purified by column chromatography (SiO2; hexane / ethyl acetate, 9:1, v / v) to give compound 108 (297 mg, 85%) as a yellow oil.

[0370] 1 H NMR (CDCl3, 400MHz): δ0.82-0.81(t, J=6.7Hz, 3H), 1.23-1.21(m, 8H), 1.41-1.71(m, 8H), 2.02(m, 1H), 2.43-2.47(m, 3H) , 3.12-3.19(m, 2H), 3.55-3.56(qui, J=6.7Hz, 1H), 4.02-4.03(m, 1H), 4.21-4.27(dd, J=2.7, 12.00Hz, 1H), 5.05(m, 1H); 13C NMR (CDCl3, 100MHz): δ14.11, 19.72, 22.73, 31.81, 25.32, 28.04, 29.02, 30.73, 32.7 1, 33.41, 34.24, 34.63, 38.55, 40.33, 56.34, 65.56, 70.73, 173.14, 173.65, 178.45.

[0371] 2-32-3,5-((2-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)octyl)oxy)-5-oxopentanoic acid [5-((2-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)octyl)oxy)-5-oxopentanoic acid] (111)

[0372] Compound 81 (204 mg, 2 equiv.) and DMAP (274 mg, 2.5 equiv.) were added to a solution of compound 110 (300 mg, 1 equiv.) and DCM (25 mL) in a reaction vessel and stirred at room temperature. The reaction mixture was vigorously stirred at room temperature for 19 hours, and the completion of the reaction was confirmed by TLC (SiO2; hexane / ethyl acetate, 8:2). The mixture was then acidified with 1 M aqueous HCl. The resulting mixture was extracted with DCM (3 × 50 mL), and the combined organic layer was washed with saturated aqueous NaCl (1 × 50 mL). The water was removed with anhydrous Na2SO4. The filtrate was then evaporated under reduced pressure. The resulting mixture was purified by column chromatography (SiO2; hexane / ethyl acetate, 9:1 to 8:2, v / v) to give compound 109 (302 mg, 75%) as a yellow liquid.

[0373] 1 H NMR (CDCl3, 400MHz): δ0.86-0.89(t, J=6.7Hz, 3H), 1.27-1.29(m, 8H), 1.45-1.70(m, 8H), 1.88-2.02(m, 5H), 2.30-2.39(t, J=8.0Hz, 2H), 2. 41-2.48(m, 8H), 3.11-3.18(m, 2H), 3.55-3.58(qui, J=6.8Hz, 1H), 3.98-4.03(m, 1H), 4.24-4.28(dd, J=2.7, 11.9Hz, 1H), 5.08-5.09(m, 1H); 13C NMR (CDCl3, 100MHz): δ14.10, 19.72, 22.79, 31.79, 29.31, 25.32, 28.01, 29.03, 30.72, 3 2.75, 33.31, 34.23, 34.62, 38.51, 40.20, 56.33, 65.55, 70.71, 173.11, 173.60, 178.43.

[0374] 2-32-4,2-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)octyl(17-hexyl-2,8,8-trimethyl-5,10,14-trioxo-6,15-dioxa-2,11-diazapentacosan-9-yl)glutarate (Compound 38)

[0375] Compound 109 (215 mg, 1.3 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (113 mg, 1.6 equiv.) and DMAP (9 mg, 0.2 equiv.) were added and the mixture was stirred vigorously under argon gas at 5°C for 10 minutes. Compound 88 (200 mg, 1 equiv.) in DCM (10 mL) was added dropwise at 5°C and the mixture was stirred at room temperature for 21 hours. After confirming the completion of the reaction by TLC (SiO2; ethyl acetate), additional DCM (50 mL) was added and the mixture was washed sequentially with saturated anhydrous NaHCO3 aqueous solution (2 × 50 mL) and saturated NaCl aqueous solution (1 × 20 mL). The obtained organic layer was dehydrated with anhydrous MgSO4, and the filtered filtrate was distilled under reduced pressure and purified by column chromatography (SiO2; ethyl acetate / hexane, 5:5 to 7:3, v / v) to obtain compound 38 (251 mg, 70%) as a yellow liquid.

[0376] 11H NMR (CDCl3, 400 MHz): δ 0.86 - 0.89 (t, J = 6.9 Hz, 9H), 1.05 - 1.06 (d, J = 6.5 Hz, 6H), 1.26 (m, 33H), 1.45 - 1.71 (m, 10H), 1.90 - 1.99 (m, 3H), 2.28 (s, 6H), 2.30 - 2.34 (t, J = 7.4 Hz, 2H), 2.38 - 2.41 (t, J = 7.2 Hz, 2H), 2.44 - 2.61 (m, 8H), 2.67 - 2.72 (m, 1H), 3.08 - 3.21 (m, 2H), 3.34 - 3.39 (m, 1H), 3.53 - 3.60 (m, 2H), 3.79 - 3.82 (d, J = 11.2 Hz, 1H), 3.96 - 4.04 (m, 4H), 4.23 - 4.26 (m, 1H), 4.85 (s, 1H), 5.06 - 5.09 (m, 1H), 7.43 - 7.46 (t, J = 5.7 Hz, 1H); 13 13C NMR (CDCl3, 100 MHz): δ 14.01, 14.08, 14.09, 20.04, 20.26, 22.02, 22.51, 22.63, 22.65, 24.56, 25.06, 26.63, 26.67, 28.69, 29.00, 29.29, 29.54, 29.58, 29.92, 30.71, 31.15, 31.59, 31.79, 31.87, 33.02, 33.04, 33.29, 33.79, 33.83, 34.57, 35.05, 37.19, 37.23, 38.46, 40.20, 45.19, 55.10, 56.30, 64.94, 67.57, 67.57, 69.57, 71.69, 76.61, 167.81, 171.66, 171.92, 172.35, 172.39, 173.12。

[0377]

Chem.

[0378] 2-33-1. O,O'-(4-((3-((9H-Fluoren-9-yl)methoxy)-3-oxopropyl)amino)-2,2-dimethyl-4-oxobutane-1,3-diyl)bis(2-(octanoyloxy)octyl)diglutarate] (112)

[0379] Compound 94 (150 mg, 1 equiv.), compound 108 (335 mg, 2.3 equiv.), EDCI·HCl (181 mg, 2.5 equiv.), and DMAP (9 mg, 0.2 equiv.) were added to a reaction vessel with DCM (50 mL) and stirred at room temperature for 13 h. After completion of the reaction was confirmed by TLC (SiO2; hexane / ethyl acetate, 9:1), additional DCM (100 mL) was added and the mixture was washed sequentially with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (1 × 200 mL). The resulting organic layer was removed with anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting mixture was purified by column chromatography (SiO2; ethyl acetate / hexane, 1:9, v / v) to give compound 112 (300 mg, 70%) as a clear liquid.

[0380] 1 H NMR (CDCl3, 400MHz): δ0.85-0.88(t, J=6.9Hz, 12H), 1.00(s, 3H), 1.04(s, 3H), 1.27(m, 36H), 1.55-1.59(m, 9H), 1.73(s, 1H), 1. 90-1.97(m, 4H), 2.26-2.31(m, 4H), 2.34-2.41(m, 6H), 2.44-2.48(t, J=7.2Hz, 3H), 2.61-2.64(t, J=5.9Hz, 2H), 3.45-3.59(m, 2 H), 3.83-3.86(m, 1H), 3.96-4.04(m, 3H), 4.18-4.23(m, 3H), 4.38-4.40(m, 2H), 4.95(s, 1H), 5.04-5.09(m, 2H), 6.67-6.70(t, J =5.9Hz, 1H), 7.29-7.33(t, J=7.4Hz, 2H), 7.39-7.42(t, J=7.4Hz, 2H), 7.57-7.58(d, J=6.4Hz, 2H), 7.76-7.77(d, J=7.5Hz, 2H); 13C NMR (CDCl3, 100MHz): δ14.03, 14.06, 19.69, 20.10, 20.87, 21.26, 22.52, 22.59, 24.88, 25 .07, 28.92, 29.03, 29.06, 30.71, 31.60, 31.65, 32.95, 33.12, 33.21, 33.40, 33.64, 34.12, 34.64, 37.36, 46.66, 64.73, 64.80, 66.70, 69.21, 71.71, 71.84, 120.08, 124.99, 127.16, 127.89, 141.29, 143.55, 143.62, 167.91, 171.57, 172.32, 172.39, 172.47, 172.65, 173.65.

[0381] 2-33-2,17-Hexyl-7,7-dimethyl-6-((5-((2-(octanoyloxy)octyl)oxy)-5-oxopentanoyl)oxy)-5,10,14,19-tetraoxo-9,15,18-trioxa-4-azahexacosanoic acid (113)

[0382] A 20% piperidine:80% DMF (v / v) solution was prepared in a reaction vessel and cooled to 0 °C. Compound 112 (300 mg) was dissolved in DMF (20 mL) in another reaction vessel and stirred for 5 min while cooling to 0 °C. Next, chilled 20% piperidine solution (30 mL) was added under argon gas and stirred for 10 min. After confirming completion of the reaction by TLC (SiO2; hexane / ethyl acetate, 5:5), 1 M aqueous HCl was added to adjust the pH to 3, and the reaction mixture was diluted with additional ethyl acetate (200 mL). The organic layer was then washed sequentially with distilled water (2 × 200 mL) and saturated aqueous NaCl (2 × 200 mL). The obtained organic layer was dehydrated using anhydrous Na2SO4, filtered, and the filtrate was distilled under reduced pressure. This was purified by column chromatography (SiO2; ethyl acetate / hexane / acetic acid, 0:1:0.1 to 5:5:0.1, v / v / v) to obtain compound 113 (238 mg, 94%) as a clear liquid.

[0383] 1H NMR(CDCl3、400MHz):δ0.84-0.87(t、J=6.5Hz、12H)、1.01(s、3H)、1.05(s、3H)、1.28(m、35H)、1.56-1.60(m、9H)、1.74(s、1H)、1.91-1.96(m、4H)、2.25-2.33(m、4H)、2.35-2.42(m、6H)、2.45-2.49(t、J=7.7Hz、3H)、2.62-2.65(t、J=5.7Hz、2H)、3.46-3.58(m、2H)、3.84-3.87(m、1H)、3.96-4.01(m、3H)、4.17-4.26(m、2H)、4.96(s、1H)、5.05-5.10(m、2H)、6.68-6.72(t、J=5.7Hz、1H); 13 C NMR(CDCl3、100MHz):δ14.00、19.82、19.87、21.03、20.87、21.23、22.48、22.54、24.84、25.02、28.86、29.00、30.65、31.55、31.60、32.86、33.06、33.21、33.34、34.10、34.76、37.28、64.78、69.24、71.76、72.05、168.02、171.67、172.53、172.65、172.79、173.63、173.82。

[0384] 2-33-3. O,O'-(2,2-Dimethyl-4-((3-(2-morpholinoethoxy)-3-oxopropyl)amino)-4-oxobutane-1,3-diyl)bis(2-(octanoyloxy)octyl)diglutarate] (Compound 39)

[0385] Compound 113 (200 mg, 1 equiv.) was placed in a reaction vessel and dissolved in DCM (30 mL). EDCI·HCl (60 mg, 1.5 equiv.) and DMAP (5 mg, 0.2 equiv.) were then added, and the reaction mixture was vigorously stirred under argon gas at 5°C for 10 minutes. Compound 101 (30 mg, 1.1 equiv.) was then added dropwise to DCM (10 mL) at 5°C, and the mixture was stirred at room temperature for 6 hours. After confirming the completion of the reaction using TLC (SiO2; ethyl acetate), an additional 100 mL of DCM was added, and the organic layer was washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (1 × 200 mL). The resulting organic layer was then dehydrated with anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The reaction mixture was then purified by column chromatography (SiO2; ethyl acetate / hexane, 7:3 to 10:0, v / v) to obtain compound 39 (172 mg, 75%) as a transparent liquid.

[0386] 1 H NMR (CDCl3, 400MHz): δ0.84-0.88(t, J=6.9Hz, 12H), 1.00(s, 3H), 1.04(s, 3H), 1.26-1.28(m, 34H), 1.57-1. 61(m, 8H), 1.89-1.99(m, 4H), 2.27-2.30(t, J=7.5Hz, 4H), 2.34-2.40(m, 6H), 2.45-2.49(m, 6H), 2.52-2.55 (t, J=6Hz, 2H), 2.58-2.61(t, J=5.8Hz, 2H), 3.44-3.56(m, 2H), 3.67-3.69(t, J=4.6Hz, 4H), 3.82-3.85(d, J =11Hz, 1H), 3.98-4.04(m, 3H), 4.16-4.26(m, 4H), 4.94(s, 1H), 5.05-5.06(m, 2H), 6.70-6.72(t, J=6Hz, 1H); 13C NMR (CDCl3, 100MHz): δ13.99, 14.02, 19.93, 19.97, 20.09, 20.85, 21.25, 22.49, 22.56, 2 4.86, 25.05, 28.88, 28.99, 29.03, 30.69, 31.57, 31.62, 32.93, 33.11, 33.20, 33.38, 33. 64, 34.10, 34.66, 37.33, 53.80, 56.98, 61.77, 64.67, 64.67, 64.70, 64.76, 66.84, 69.18, 71.70, 71.84, 76.63, 167.81, 171.53, 172.29, 172.34, 172.39, 172.50, 173.46, 173.49.

[0387] [ka]

[0388] 2-34-1,4-((3-((9H-Fluoren-9-yl)methoxy)-3-oxopropyl)amino)-3-hydroxy-2,2-dimethyl-4-oxobutyl(2-hexyldecyl)glutarate] (114)

[0389] Compound 108 (1.07 g, 1.1 equiv.), EDCI·HCl (0.72 g, 1.5 equiv.), and DMAP (46 mg, 0.15 equiv.) were placed in a reaction vessel, and DCM (20 mL) was added. The mixture was then vigorously stirred under argon gas at room temperature for 20 minutes. Compound 94 (1 g, 1 equiv.) was dissolved in DCM (20 mL) in a separate reaction vessel and cooled to 5°C. This mixture was then added dropwise to the previously prepared reaction mixture under argon gas at 5°C for 30 minutes. After stirring at room temperature for 9 hours, the reaction was confirmed to be complete by TLC (SiO2; hexane / ethyl acetate, 6:4). The solvent was then removed by distillation under reduced pressure. An additional 50 mL of DCM was added, and the mixture was washed with saturated aqueous NaHCO3 (2 × 100 mL) and saturated aqueous NaCl (1 × 100 mL). The organic layer was collected and dehydrated with anhydrous MgSO4. The filtered filtrate was concentrated by distillation under reduced pressure and then purified by column chromatography (SiO2; EtOAc / hexane, 1:9 to 4:6, v / v) to give compound 114 (1.49 g, 72%) as a clear liquid.

[0390] 1 H NMR(CDCl3、400MHz):δ0.85-0.89(t、J=6.7Hz、9H)、1.04(s、3H)、1.25(m、25H)、1.60(bs、1H)、1.92-1.98(m、2H)、2.34-2.47(m、4H)、2.60-2.63(t、J=5.9Hz、2H)、3.50-3.55(m、2H)、3.60-3.61(d、J=5.8Hz、1H)、3.73-3.76(d、J=11、1H)、3.84-3.86(d、J=5.7、1H)、3.95-3.96(d、J=5.6、2H)、4.16-4.21(m、2H)、4.39-4.42(m、2H)、7.00-7.03(t、J=11.7、1H)、7.29-7.32(t、J=7.4、2H)、7.38-4.41(t、J=7.4、2H)、7.55-7.57(d、J=7.4、2H)、7.74-7.76(d、J=7.52、2H); 13 C NMR(CDCl3、100MHz):δ14.09、14.10、19.57、20.25、21.37、22.64、22.66、26.64、26.69、29.30、29.55、26.59、29.94、31.22、31.80、31.89、33.37、33.38、34.08、34.62、37.26、38.62、46.68、68.57、67.46、70.56、74.65、102.07、124.94、127.15、127.85、141.30、143.54、171.92、172.15、173.25、173.40。

[0391] 2-34-2,2-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)octyl(1-(9H-fluoren-9-yl)-19-hexyl-9,9-dimethyl-3,7,12,16-tetraoxo-2,11,17-trioxa-6-azaheptacosan-8-yl)glutarate] (115)

[0392] Compound 114 (1.49 g, 1 equiv.) was dissolved in DCM (50 mL) in a reaction vessel, followed by the addition of compound 111 (1.08 g, 1.2 equiv.), EDCI·HCl (0.58 g, 1.5 equiv.), and DMAP (37 mg, 0.15 equiv.). The mixture was stirred at room temperature for 13 hours. After completion of the reaction was confirmed by TLC (SiO2; hexane / ethyl acetate, 7:3), an additional 100 mL of DCM was added. The organic layer was washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (1 × 200 mL). The combined organic layer was then removed with anhydrous MgSO4, filtered, and concentrated by vacuum distillation. The resulting reaction mixture was purified by column chromatography (SiO2; ethyl acetate / hexane, 1:9 to 3:7, v / v) to give compound 115 (1.41 g, 60%) as a pale yellow liquid.

[0393] 1 H NMR (CDCl3, 400MHz): δ0.85-0.89(t, J=3.2Hz, 9H), 1.00(s, 3H), 1.04(s, 3H), 1.25(m, 34H), 1.44-1.65(m, 12H), 1.84-1.97(m, 5H), 2.27- 2.30(t, J=6.5Hz, 2H), 2.32-2.48(m, 9H), 2.60-2.63(t, J=6.0Hz, 2H), 3.04-3.17(m, 2H), 3.44-3.58(m, 3H), 3.82-3.85(t, J=12.0Hz, 1H) , 3.94-4.01(m, 3H), 4.02-4.05(t, J=11.0Hz, 1H), 4.18-4.25(m, 2H), 4.37-4.39(d, J=7.2Hz, 2H), 4.94-4.95(m, 1H), 5.05(bs, 1H), 6.68- 6.71(t, J=5.56Hz, 1H), 7.28-7.32(t, J=7.7Hz, 2H), 7.37-7.41(t, J=7.4Hz, 2H), 7.56-7.58(d, J=7.4Hz, 2H), 7.74-7.76(d, J=7.4Hz, 2H); 13C NMR (CDCl3, 100MHz): δ14.03, 14.09, 14.11, 19.95, 19.99, 20.11, 20.85, 21.32, 22.21, 22.62, 22.65, 24.56, 25 .05, 26.62, 26.66, 28.67, 28.99, 29.27, 29.53, 29.57, 29.92, 30.63, 31.21, 31.58, 31.78, 31.87, 33.20, 33.33, 33.63, 33.82, 34.55, 34.66, 37.25, 37.34, 38.44, 40.18, 46.66, 56.29, 64.87, 66.67, 67.29, 69.17, 71.75, 76.69, 120.06, 124.99, 127.15, 127.85, 141.26, 143.56, 167.91, 171.55, 172.30, 172.40, 172.56, 173.03, 173.06.

[0394] 2-34-3,21-(1,2-Dithiolan-3-yl)-15-hexyl-6-(1-((5-((2-hexyldecyl)oxy)-5-oxopentanoyl)oxy)-2-methylpropan-2-yl)-5,8,12,17-tetraoxo-7,13,16-trioxa-4-azahenicosanoic acid (116)

[0395] A 20% piperidine:80% DMF (v / v) solution was prepared in a reaction vessel and cooled to 0 °C. Compound 115 (1.41 g) was dissolved in DMF (20 mL) in another reaction vessel and stirred at 0 °C for 5 minutes. The 20% piperidine solution (60 mL) was added to the reaction mixture under argon gas and stirred for 5 minutes. After confirming completion of the reaction by TLC (SiO2; ethyl acetate), the mixture was acidified to pH 3 with 1 M aqueous HCl. The reaction mixture was diluted with ethyl acetate (200 mL) and washed with distilled water (2 × 200 mL) and saturated aqueous NaCl (2 × 200 mL). The organic layer was collected, removed with anhydrous Na2SO4, filtered, and the filtrate was concentrated by vacuum distillation. It was then purified by column chromatography (SiO2; ethyl acetate / MeOH, 10:0 to 9:1, v / v) to give compound 116 (0.79 g, 66%) as a yellow liquid.

[0396] 1H NMR(CDCl3、400MHz):δ0.85-0.88(t、J=3.1Hz、9H)、1.01(s、3H)、1.10(s、3H)、1.24(m、35H)、1.45-1.67(m、12H)、1.85-1.92(m、5H)、2.28-2.32(t、J=6.4Hz、2H)、2.31-2.47(m、9H)、2.61-2.62(t、J=6.1Hz、2H)、3.05-3.18(m、2H)、3.45-3.59(m、3H)、3.85-3.88(t、J=9.0Hz、1H)、3.95-4.02(m、3H)、4.03-4.07(t、J=11.0Hz、1H)、4.21(d、J=7.1Hz、1H)、4.90-4.93(m、1H)、5.06(bs、1H)、6.69-6.72(t、J=5.62Hz、1H); 13 C NMR(CDCl3、100MHz):δ14.01、14.10、19.94、20.01、20.13、20.84、21.35、22.25、22.63、22.66、24.57、25.04、26.63、26.68、28.68、20.03、29.28、29.54、29.55、29.91、30.64、31.22、31.57、31.79、31.87、33.21、33.34、33.64、33.81、34.56、34.67、37.23、37.33、38.45、40.19、56.27、66.66、67.32、69.19、71.78、76.71、167.96、171.57、172.33、172.42、172.57、173.02、173.09。

[0397] 2-34-4,2-((5-(1,2-Dithiolan-3-yl)pentanoyl)oxy)octyl(20-hexyl-10,10-dimethyl-1-morpholino-4,8,13,17-tetraoxo-3,12,18-trioxa-7-azaoctacosan-9-yl)glutarate (Compound 40)

[0398] Compound 116 (434 mg, 1 equiv.) was dissolved in DCM (30 mL) and EDCI·HCl (126 mg, 1.5 equiv.) and DMAP (11 mg, 0.2 equiv.) were added to a reaction vessel. The mixture was stirred vigorously under argon at 5 °C. Compound 101 (70 mg, 1.2 equiv.) in DCM (10 mL) was then added dropwise at 5 °C and stirred at room temperature for 7 h. After confirming the completion of the reaction by TLC (SiO2; ethyl acetate), an additional 100 mL of DCM was added and the mixture was washed with saturated aqueous NaHCO3 (2 × 200 mL) and saturated aqueous NaCl (1 × 200 mL). The organic layer was collected and dehydrated with anhydrous MgSO4, and the filtered filtrate was concentrated by distillation under reduced pressure. The resulting reaction mixture was purified by column chromatography (SiO2; ethyl acetate / hexane, 5:5 to 10:0, v / v) to give compound 40 (378 mg, 78%) as a yellow liquid.

[0399] 1 H NMR (CDCl3, 400MHz): δ0.80-0.83(t, J=5.8Hz, 9H), 0.96(s, 3H), 0.83(s, 3H), 1.21(m, 30H), 1.39-1.43(m, 3 H), 1.51-1.65(m, 8H), 1.84-1.92(m, 5H), 2.25-2.36(m, 8H), 2.38-2.44(m, 7H), 2.47-2.50(t, J=5.9Hz, 2H), 2.54-2.57(t, J=5.8Hz, 2H), 3.01-3.14(m, 2H), 3.39-3.52(m, 3H), 3.62-3.64(t, J=4.2Hz, 4H), 3.77-3.80( m, 1H), 3.91-4.00(m, 4H), 4.01-4.20(m, 3H), 4.88-4.90(m, 1H), 5.01(bs, 1H), 6.67-6.69(t, J=5.64Hz, 1H); 13C NMR (CDCl3, 100MHz): δ14.02, 14.09, 19.94, 19.97, 20.08, 20.87, 21.28, 22.50, 22.61, 22.64, 24.56, 25.06 , 26.61, 26.65, 28.69, 28.99, 29.27, 29.52, 29.57, 29.91, 30.69, 31.19, 31.58, 31.77, 31.86, 32.95, 33.02, 33.19, 33.32, 33.64, 33.83, 34.56, 34.65, 37.24, 37.32, 38.45, 40.20, 53.80, 56.29, 56.97, 61.78, 64.87, 66.84, 67.30, 69.16, 71.63, 71.77, 76.67, 167.80, 171.56, 172.32, 172.42, 172.58, 172.99, 173.05, 173.12.

[0400] [Example 3] Synthesis of butyllithocholic acid

[0401] Lithocholic acid and n-butanol were placed in a reaction vessel, and HCl solution (35-37% aqueous solution, 1 equivalent) was added. The mixture was then stirred at room temperature for 24 hours. After distilling the alcohol using a rotary evaporator, the mixture was redissolved in DCM and impurities were extracted using saturated aqueous NaHCO3 and saturated brine. The DCM layer was then dehydrated using anhydrous Na2SO4, filtered, and the filtered solution was dried using a rotary evaporator and purified by column chromatography (SiO2, DCM / MeOH, 19:1-49:1 volume ratio). This yielded the lithocholic acid derivative compound (92%).

[0402] MS(ESI-MS) calculation for C 28 H 46 NO2[M+H] + 428.3523,found 428.3525.

[0403] Example 4: Synthesis of trehalose-based lipids

[0404] Trehalose dihydrate (200 mg, 0.53 mmol) was dissolved in pyridine (5 mL), followed by the addition of TBTU (421 mg, 1.3 mmol), DIPEA (320 μL, 1.2 mmol), and oleic acid (370 μL, 1.2 mmol), in that order. The mixture was then backfilled with argon and stirred at room temperature for 20 hours. The solvent was removed using a rotary evaporator, and the solid was purified by column chromatography (SiO2, MeOH / EtOAc 2:98->10:90) and dried. The dried solid was washed several times with EtOAc and then thoroughly dried to yield 287 mg (62%) of 6,6'-trehalose dioleate.

[0405] 1 H NMR (400MHz, MeOD) δ5.38(t, J=4.5Hz, 4H), 5.08(d, J=3.8Hz, 2H), 4.39(dd, J=2.2, 11.9 Hz, 2H), 4.23(dd, J=5.1, 11.9Hz, 2H), 4.04(ddd, J=2.1, 5.3, 10.2Hz, 2H), 3.81(dd, J=9. 4, 9.6Hz, 2H), 3.50(dd, J=3.6, 9.6Hz, 2H), 3.35(dd, J=8.8, 10.0Hz, 2H), 2.04-2.09(m, 8 H), 2.37(t, J=7.4Hz, 4H), 1.61-1.67(m, 4H), 1.29-1.41(m, 40H), 0.93(t, J=6.8Hz, 6H); 13 C NMR (100MHz, MeOD) δ174.0, 129.5, 129.4, 93.8, 73.2, 71.8, 70.5, 70.1, 63.0, 33 .7, 31.7, 29.5, 29.4, 29.2, 29.1, 29.0, 28.9, 28.8, 28.8, 26.8, 24.7, 22.4, 13.1.

[0406] [Example 5] Preparation of RNA platform nucleic acid molecules

[0407] We created an RNA platform with an IRES element derived from encephalomyocarditis virus (EMCV) and a nucleic acid sequence encoding Renilla luciferase (R / L), a known reporter gene, inserted as a target sequence. We designed a template DNA and used the IVT process to create a single-stranded RNA platform nucleic acid molecule.

[0408] We constructed an RNA platform containing the Cap1 structure and a nucleic acid sequence encoding influenza virus hemagglutinin (HA) as a target sequence. We designed a template DNA and used the IVT process to generate a single-stranded RNA platform nucleic acid molecule.

[0409] [Example 6] Preparation of lipid nanoparticles

[0410] Lipid nanoparticles are prepared by mixing a 0.625 mg / mL RNA solution (50 mM sodium citrate buffer, 110 mM NaCl, pH 4.0) and a lipid mixture solution (Ethanol) using a laboratory mixer and emulsifier (NanoAssemblr Spark, Precision Nanosystems, Inc.), followed by solvent exchange with saline or PBS using a centrifugal filter tube (UFC5010, Amicon).

[0411] Specifically, the contents were prepared as shown in Tables 1 to 3 below.

[0412] [Table 1]

[0413] [Table 2-1] [Table 2-2]

[0414] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7]

[0415] [Experimental Example 1] Evaluation of transduction efficiency through confirmation of protein expression level

[0416] mRNA-LNP was injected ID into the ears of ICR rats at an RNA standard of 5 μg / 20 μl to confirm the presence or absence of expression.

[0417] The experimental procedure is as follows:

[0418] Each experimental group was prepared with a volume of 5 μg / 20 μl of RNA, which was then injected into both ears of the rats via ID injection. Expression of the target protein was then confirmed during the time period when it was most expressed. In the case of R / L, where the experiment was actually conducted, expression and retention were confirmed during two time periods: 6 hours and 24 hours.

[0419] The specific experimental process is as follows:

[0420] After anesthesia using an individual respiratory anesthesia machine, the administration drug according to the experimental conditions was injected via insulin syringe. Thereafter, the animals were euthanized using CO2 at the appropriate time points, and their ears were then cut. The cut ears were immersed in 300 μl of 1× Renilla lysis buffer and thoroughly crushed using scissors and a homogenizer. 20 μl of the mixture was then transferred to a white 96-well plate, and 100 μl of Promega's Renilla luciferase assay substrate was added. Luminescence was measured to determine whether expression was present or not.

[0421] Figure 1 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 127, 130) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. As shown in Figure 1, comparing LNP 98 and LNP 127, protein expression efficiency was maintained even when Formula 10 was used as the helper lipid. Furthermore, comparing LNP 98 and LNP 130, a certain degree of expression efficiency was maintained even when Formula 10 was used as the additive, increasing the ratio of helper lipid.

[0422] Figure 2 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 131, 132, and 133) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. As shown in Figure 2, comparing LNP 128 and LNP 133, protein expression efficiency was maintained even when Formula 10 was used as the helper lipid. Furthermore, comparing LNP 131 and LNP 132, there was no significant difference between Formula 10 mixed with DOPE and when used alone, confirming that reducing the ionizable lipid content by half did not dramatically affect protein expression efficiency.

[0423] Figure 3 shows the analysis of the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 136, 138, and 139) with a novel helper lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 3 shows that when comparing LNP 128 and LNP 139, substitution of Formula 11 as the helper lipid maintained protein expression efficiency at 6 hours and even increased it at 24 hours. Furthermore, LNP 136 and LNP 138 containing Formula 10 and Formula 11 also effectively functioned as helper lipids, confirming their potential as substitutes.

[0424] Figure 4 shows the analysis of the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 109, 111, and 114) with novel cholesterol-substituting compositions containing Renilla luciferase (R / L) mRNA into rat ears. Comparing LNP 98 and LNP 109 in Figure 4, there was no significant difference in protein expression efficiency when cholesterol was replaced with vitamin A, confirming that vitamin A can be used to replace cholesterol. Furthermore, LNP 111 and LNP 114 confirmed that protein expression efficiency decreased when vitamin E or vitamin K was used.

[0425] Figure 5 shows the analysis of the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 110, 115, 117, and 118) with novel cholesterol-substituting compositions containing Renilla luciferase (R / L) mRNA into rat ears. In Figure 5, comparing LNP 98 and LNP 110, the protein expression efficiency decreased when cholesterol was partially mixed with butyllithocholic acid rather than completely substituted for it due to the solubility of coenzyme Q10. Furthermore, in LNP 115 and 117, vitamin K2 and vitamin C palmitate decreased the protein expression efficiency after cholesterol substitution. Furthermore, in LNP 118, when β-carotene was substituted, there was no significant difference in the protein expression efficiency compared to LNP 98, confirming its potential as a cholesterol substitute.

[0426] Figure 6 shows the analysis of the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 119, 123) with novel cholesterol-substituting compositions containing R / L mRNA into rat ears. Comparing LNP 98 and LNP 119 in Figure 6, the ratio of coenzyme Q10 was changed depending on the solubility of coenzyme Q10. However, expression efficiency was slightly lower than with butyllithocholic acid, and optimal ratios must be explored. Comparing LNP 109 (see Figure 4) and LNP 123, retinol, which is a form of vitamin A (retinoic acid) reduced to alcohol, was not able to stably substitute for cholesterol, unlike vitamin A.

[0427] Figure 7 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 168) with a novel cholesterol-replacing composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 7 shows that there was no significant difference in protein expression efficiency between LNP 98 and LNP 168, confirming that the use of luteolin can maintain a certain degree of structure. Luteolin is a flavonoid and a phytochemical also known as vitamin P, known for its antioxidant properties, scavenging reactive oxygen species, and regulating metabolism and the immune system.

[0428] Figure 8 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 200) with a novel cholesterol-substitute composition containing Renilla luciferase (R / L) mRNA into rat ears. Comparing LNP 126 and LNP 200 in Figure 8, 3-oxo-5β-cholanoic acid, when used in place of butyllithocholic acid, further increased the efficiency of protein expression. It is believed that 3-oxo-5β-cholanoic acid acts as an inhibitor of ROR-γt, which is involved in immune activation, and thus prevents mRNA degradation by the in vivo immune system prior to protein expression.

[0429] In conclusion, vitamin A, β-carotene, luteolin, 3-oxo-5β-cholanic acid, etc. showed outstanding effects as alternatives to structure-retaining compounds.

[0430] Figure 9 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 146 and 147) with novel ionizable lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. Figure 9 shows that when compounds 12 and 13 were introduced, respectively, instead of the ionizable lipid MC3 in LNP 98 (LNP 146 and LNP 147), the protein expression efficiency was slightly reduced in both cases compared to LNP 98.

[0431] Figure 10 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 152, 153) with a novel ionizable lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 10 shows that when compound 24 was introduced instead of the ionizable lipid MC3 in LNP 98 (LNP 152), the protein expression efficiency increased compared to LNP 98. However, when compound 24 was introduced instead of SM-102 in LNP 128 (LNP 153), the protein expression efficiency decreased compared to LNP 128.

[0432] Figure 11 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 159, 160) with a novel ionizable lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 11 shows that when compound 16 was introduced instead of the ionizable lipid DLin-MC3-DMA in LNP 98 (LNP 159), the initial protein expression efficiency increased slightly compared to LNP 98, but the persistence of expression decreased. When compound 16 was introduced instead of SM-102 in LNP 128 (LNP 160), the protein expression efficiency decreased slightly compared to LNP 128.

[0433] Figure 12 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 166, 167) with a novel ionizable lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 12 shows that when compound 14 was introduced instead of the ionizable lipid MC3 in LNP 98 (LNP 166), the protein expression efficiency remained similar at 6 hours compared to LNP 98, but was significantly reduced at 24 hours. When compound 14 was introduced instead of SM-102 in LNP 128 (LNP 167), the protein expression efficiency was significantly reduced compared to LNP 128.

[0434] Figure 13 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 178, 179, 181, and 182) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. In Figure 13, when compound 16 (LNP 178) and compound 14 (LNP 179) were used instead of SM-102, the ionized lipid of LNP 139 using compound 10 as the helper lipid, LNP 178 showed a slight decrease in protein expression efficiency, while LNP 179 showed a significant decrease in protein expression efficiency. When compound 16 (LNP 181) and compound 14 (LNP 182) were used instead of SM-102, the ionizable lipid of LNP 138 using compound 11 as a helper lipid, LNP 181 maintained its expression efficiency at 6 hours but showed a slight decrease in expression efficiency at 24 hours.

[0435] Figure 14 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after the injection (id) of lipid nanoparticles (LNP 185, 187) with novel ionizable lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. In Figure 14, to optimize the LNP composition using compound 16 as the ionizable lipid, the butyllithocholic acid in LNP 181 was replaced with quercetin (LNP 185) and 3-oxo-5β-cholanic acid (LNP 187). As a result, the protein expression efficiency of LNP 187 significantly increased. Quercetin is a plant flavonoid known to have excellent antioxidant properties and to have neuroprotective, cardioprotective, and anti-inflammatory effects; however, the protein expression efficiency did not significantly increase. Among cholesterol substitutes, 3-oxo-5β-cholanic acid showed the highest transfer efficiency. By using 3-oxo-5β-cholanic acid instead of butyllithocholic acid in LNP 181, we confirmed that protein expression efficiency and durability were further increased.

[0436] Figure 15 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 192, 193) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. Figure 15 shows that when ionized lipids LNP 126 and LNP 128 were introduced into compound 15 instead of SM-102 (LNP 192 and LNP 193), the protein expression efficiency was significantly reduced compared to LNP 126 and LNP 128, respectively.

[0437] Figure 16 shows the results of analyzing the amount of Renilla luciferase (R / L) mRNA expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 205, 208, 209, and 210) with novel ionized lipid compositions. Figure 16 shows that LNP 205, which replaces the helper lipid of LNP 167 with Compound 11 and cholesterol with 3-oxo-5β-cholanic acid, significantly reduced protein expression efficiency. LNP 208, which alters the composition of LNP 187, slightly increased protein expression efficiency. In the case of LNP 209, in which 3-oxo-5β-cholanic acid in LNP 208 was changed to coenzyme Q10, the protein expression efficiency decreased. In the case of LNP 210, in which the helper lipid in LNP 160 was replaced with compound 11 and cholesterol was replaced with 3-oxo-5β-cholanic acid, the protein expression efficiency also decreased.

[0438] As a result of explaining Figures 1 to 16, among the lipid nanoparticle compositions of the present invention, when the lipid nanoparticle composition contained ionized lipid compound 16, the helper lipid was compound 11, the cholesterol was 3-oxo-5β-cholanic acid, and the PEG-lipid was DMG-PEG, (LNP 187) showed the highest protein expression efficiency.

[0439] Figure 17 shows the analysis of Renilla luciferase (R / L) mRNA expression levels 6 and 24 hours after intravenous injection (id) of lipid nanoparticles (LNP 211, 212, 213, 214, 216, and 217) containing novel ionized lipid compositions. Figure 17 shows that when the ionized lipids LNP 126 and LNP 128 were incorporated into compound 18 instead of SM-102 (LNP 211 and LNP 212), protein expression efficiency was slightly reduced compared to LNP 126. In particular, LNP 211 maintained some expression efficiency at 6 hours but significantly decreased at 24 hours. Comparing LNP 211 with LNP 213 and LNP 214, the substitution of butyllithocholic acid for 3-oxo-5β-cholanic acid resulted in a greater increase in protein expression efficiency at 24 hours. However, when compound 14 was used as the ionizable lipid, the protein expression efficiency was significantly reduced when butyllithocholic acid was replaced with 3-oxo-5β-cholanic acid, as compared with LNP 166, LNP 216, and LNP 217.

[0440] Figure 18 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 224, 225, and 226) with novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. Figure 18 illustrates that when ionized lipids LNP 126 and LNP 128 were introduced into compound 17 instead of SM-102 (LNP 224 and LNP 225), the protein expression efficiency was slightly reduced compared to LNP 126.

[0441] Figure 19 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after the injection (id) of lipid nanoparticles (LNP 227, 228, 229, 230, 231, and 232) containing novel ionized lipid compositions containing Renilla luciferase (R / L) mRNA into rat ears. Figure 19 illustrates that when the ionized lipids of LNP 126 and LNP 128 were introduced into compound 19 instead of SM-102 (LNP 227 and LNP 228), the protein expression efficiency was significantly reduced compared to LNP 126. When the ionized lipid of LNP 187 was introduced into compound 19 instead of compound 16, the protein expression efficiency did not increase significantly. On the other hand, when the ionized lipids LNP 126 and LNP 128 were introduced into compound 20 instead of SM-102 (LNP 230 and LNP 231), the protein expression efficiency was higher than that of LNP 126. In particular, LNP 230 showed a significant increase in expression efficiency at both 6 and 24 hours. Furthermore, when the ionized lipid LNP 187 was introduced into compound 20 instead of compound 16 (LNP 232), the protein expression efficiency at 6 hours was further increased, demonstrating that optimization using 3-oxo-5β-cholanic acid is well suited to this.

[0442] Figure 20 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 256 and 257) with a novel ionizable lipid composition containing Renilla luciferase (R / L) mRNA into rat ears. Figure 20 illustrates that when compound 23 was introduced into LNP 126 and LNP 128 (LNP 256 and LNP 257), respectively, instead of the ionizable lipid SM-102, the protein expression efficiency was significantly reduced compared to LNP 126 and LNP 128.

[0443] Figure 21 shows the results of analyzing the amount of Renilla luciferase (R / L) mRNA expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 278, 279, 280, and 281) containing novel ionizable lipid compositions into rat ears. Figure 21 shows that the protein expression efficiency was significantly reduced when compound 21 (LNP 278) and compound 22 (LNP 280) were introduced into LNP 126 instead of the ionizable lipid SM-102, and when compound 21 (LNP 279) and compound 22 (LNP 281) were introduced into LNP 128 instead of the ionizable lipid SM-102.

[0444] 10 to 21, it can be seen that among the compounds represented by Chemical Formula 4, LNP 230, LNP 231, and LNP 232, which use Compound 20 as an ionized lipid, have the highest expression efficiency.

[0445] Figure 22 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after the injection (id) of lipid nanoparticles (LNP 291, 292, 293, 294, and 295) containing novel ionized lipids containing Renilla luciferase (R / L) mRNA into rat ears. As shown in Figure 22, when the ionized lipids LNP 126 and LNP 128 were introduced into compound 25 instead of SM-102 (LNP 291 and LNP 292), the protein expression efficiency decreased compared to LNP 126. When the ionized lipid LNP 187 was introduced into compound 25 instead of compound 16 (LNP 293), the protein expression efficiency did not increase significantly, but it did increase slightly compared to LNP 291. On the other hand, when the ionizable lipids LNP 126 and LNP 128 were introduced into compound 26 instead of SM-102 (LNP 294 and LNP 295), the protein expression efficiency was maintained or slightly increased compared to LNP 126. Furthermore, when the ionizable lipid LNP 187 was introduced into compound 26 instead of compound 16 (LNP 296), the protein expression efficiency was significantly increased compared to LNP 294, indicating that optimization using 3-oxo-5β-cholanic acid is well suited to this.

[0446] Figure 23 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 310, 311, 312, 313, 314, and 315) containing novel cholesterol-replacement compositions containing Renilla luciferase (R / L) mRNA into rat ears. Figure 23 illustrates that when the ionized lipids of LNP 126 and LNP 128 were introduced into compound 27 instead of SM-102 (LNP 310 and LNP 311), the protein expression efficiency was reduced compared to LNP 126. When the ionized lipid of LNP 187 was introduced into compound 27 instead of compound 16 (LNP 312), the protein expression efficiency was slightly reduced compared to LNP 310. On the other hand, when the ionizable lipids LNP 126 and LNP 128 were introduced into compound 28 instead of SM-102 (LNP 313 and LNP 314), the protein expression efficiency decreased compared to LNP 126. Furthermore, when the ionizable lipid LNP 187 was introduced into compound 28 instead of compound 16 (LNP 315), the protein expression efficiency decreased compared to LNP 313. It was found that when compound 27 and compound 28 were used as the ionizable lipid, optimization using compound 11 and 3-oxo-5β-cholanic acid was not suitable.

[0447] 22 and 23, it can be seen that among the compounds represented by Chemical Formula 8, LNP 295, which is a lipid nanoparticle containing Compound 26 as an ionized lipid, has the highest expression efficiency.

[0448] Figure 24 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after the intravenous injection (id) of lipid nanoparticles (LNP 323, 324, 325, 326, 327, and 328) containing novel ionizable lipids containing Renilla luciferase (R / L) mRNA into rat ears. Figure 24 shows that when compound 29 was introduced instead of the ionizable lipid SM-102 in LNP 126 and LNP 128 (LNP 323 and LNP 324), the protein expression efficiency was significantly reduced. On the other hand, when compound 30 was introduced instead of the ionizable lipid SM-102 in LNP 126 and LNP 128 (LNP 325 and LNP 326) or compound 31 was introduced instead (LNP 327 and LNP 328), the protein expression efficiency was slightly reduced.

[0449] Figure 25 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after the injection (id) of lipid nanoparticles (LNP 329, 330, 331, 332, 333, 334) containing novel ionizable lipids containing Renilla luciferase (R / L) mRNA into rat ears. Figure 25 illustrates that when compound 32 was introduced into LNP 126 and LNP 128 instead of the ionizable lipid SM-102, the protein expression efficiency was slightly reduced (LNP 329 and LNP 330). When compound 35 was introduced into LNP 126 and LNP 128 (LNP 331 and LNP 332) instead of the ionizable lipid SM-102, and when compound 36 was introduced (LNP 333 and LNP 334), the protein expression efficiency at 6 hours was particularly high in LNP 331 containing compound 35 and LNP 334 containing compound 36, which increased by 40% and 20%, respectively, compared to LNP 126.

[0450] Figure 26 shows the results of analyzing the amount of Renilla luciferase (R / L) expressed 6 and 24 hours after injection (id) of lipid nanoparticles (LNP 335, 336, 337, and 338) containing novel ionizable lipids containing Renilla luciferase (R / L) mRNA into rat ears. Figure 26 shows that when compound 33 (LNP 335 and LNP 336) and compound 34 (LNP 337 and LNP 338) were introduced instead of the ionizable lipid SM-102 in LNP 126 and LNP 128, respectively, the protein expression efficiency at 6 hours was maintained or slightly decreased compared to LNP 126.

[0451] Referring to Figures 24 to 26, it was confirmed that among the compounds represented by Chemical Formula 6, LNP 331, which contains Compound 35 as an ionizable lipid, and LNP 334, which contains Compound 36 as an ionizable lipid, exhibited the highest protein expression efficiency.

[0452] Figure 27 shows the analysis of Renilla luciferase (R / L) mRNA expression 6 hours after injection (id) of lipid nanoparticles (LNP 412, 417, 422, 425) containing novel ionizable lipids. Figure 27 shows that when compound 37, compound 38, compound 39, or compound 40 was introduced into LNP 128 instead of the ionizable lipid SM-102 (LNP 412, 417, 422, 425), the protein expression efficiency was slightly reduced. However, when compound 37 (LNP 412) was introduced into LNP 128 instead of compound 26 (LNP 295), the protein expression efficiency was similar, confirming that the additional ester bond introduced into compound 37 does not significantly affect mRNA encapsulation compared to compound 26.

[0453] [Experimental Example 2] HA-specific antibody immune response increased by mRNA-formulated LNP

[0454] To investigate the effect of LNPs on antibody production, mRNA encoding HA, a surface antigen of the influenza virus, was encapsulated in LNPs, and 6-week-old BALB / c rats were immunized intramuscularly with 10 μg of mRNA. Two weeks after the primary or secondary immunization, antibody responses were observed through IgG1 and IgG2a levels against HA-specific antibodies in the blood using ELISA.

[0455] The specific experimental process is as follows:

[0456] The drugs that matched the experimental conditions were injected using an insulin syringe. Thereafter, blood samples were collected using a respiratory anesthesia machine on the appropriate dates. Serum was separated from the collected blood and the experiment was then carried out.

[0457] Figure 28 shows the results of intramuscular immunization of rats with formulations of mRNA encoding influenza virus surface antigens (LNP 126, 181, 187, and 200). Two weeks after the primary immunization (a-b) and secondary immunization (c-d), the HA-specific antibody response in the blood was measured through IgG1 (a, c) and IgG2a (b, d) levels. Figure 28 shows that LNP 181, LNP 187, and LNP 200 had reduced IgG1 and IgG2a levels compared to LNP 126, confirming their insignificant immune effects. In particular, LNP 187 demonstrated significantly reduced immune effects despite increased protein expression efficiency compared to LNP 126 (Figure 14), confirming its potential as a nucleic acid therapeutic agent that can be delivered without side effects through human immune activation.

[0458] Figure 29 shows the results of intramuscular immunization of rats with formulations of mRNA encoding influenza virus surface antigens (LNP 126, 128, 230, 231, and 232), followed by monitoring the HA-specific antibody response in the blood through IgG1 (a, c) and IgG2a (b, d) levels two weeks after the primary immunization (a-b) and secondary immunization (c-d). Figure 29 shows that, compared to LNP 126 and LNP 128, LNP 230, LNP 231, and LNP 232 showed reduced IgG1 and IgG2a levels in the primary immunization, indicating a less significant immune effect. Even in the secondary immunization, there was significant inter-individual variability, and IgG2a levels remained lower than those of LNP 126 and LNP 128. In particular, LNP 231 reduces the levels of IgG1 and IgG2a after both the primary and secondary immunizations, and therefore has potential for use as a nucleic acid therapeutic agent that can be delivered without side effects based on the activation of the human immune system.

[0459] Figure 30 shows the results of intramuscular immunization of rats with formulated mRNA encoding influenza virus surface antigens (LNP 128, 295), followed by monitoring the HA-specific antibody response in the blood through IgG1 (a, c) and IgG2a (b, d) levels two weeks after the primary immunization (a-b) and secondary immunization (c-d). Regarding Figure 30, LNP 295 increased IgG1 and IgG2a levels to a similar extent as LNP 128 in both the primary and secondary immunizations, confirming its potential as an mRNA vaccine. This is consistent with the results of Figure 29, where blood MCP-1 levels were significantly increased compared to the positive control, LNP 128.

[0460] [Experimental Example 3] Increased MCP-1 protein expression by HA-encoding mRNA-formulated LNP

[0461] Monocyte chemoattracted protein-1 (MCP-1) is a type of chemokine that induces monocyte migration and adhesion to vascular endothelial cells and is known to play an important role in immune responses. To confirm the effect of LNP on immune responses, R / L-encoding mRNA was encapsulated in LNP, and the mRNA-LNP was injected into the ear of ICR rats via ID at 5μg / 20μl of RNA. Six hours later, blood was collected and the concentration was measured by ELISA.

[0462] The specific experimental process is as follows:

[0463] The appropriate drug for the experiment was injected via insulin syringe. Six hours later, blood was collected using a respiratory anesthesia machine. Serum was separated from the collected blood and the experiment was carried out. Expression levels were compared using Invitrogen's MCP-1 mouse uncoated ELISA kit.

[0464] Figure 31 shows the results of measuring MCP-1 concentrations in blood collected 6 hours after intravenous administration of formulations (LNP 126, 159, 211, 224, 227, 230, 291, 294, 310, 313, 323, 325, 327, 329, 331, 333, 335, and 337) encoding Renilla luciferase (R / L) to ICR rats. Figure 31 shows that LNP 313 and LNP 337 increased blood MCP-1 concentrations compared to the positive control, LNP 126, while LNP 291 and LNP 327 showed similar blood MCP-1 concentrations compared to LNP 126. Among them, LNP 327 showed a high MCP-1 concentration while maintaining a certain degree of protein expression efficiency, and is therefore expected to be useful as an mRNA vaccine.

[0465] Figure 32 shows the results of measuring MCP-1 concentrations in blood collected 6 hours after intravenous administration of formulations (LNP 128, 160, 212, 225, 228, 231, 292, 295, 311, 314, 324, 326, 328, 330, 332, 334, 336, and 338) encoding Renilla luciferase (R / L) to ICR rats. Figure 32 shows that LNP 295 significantly increased blood MCP-1 concentrations compared to the positive control, LNP 128. Since LNP 212, LNP 231, LNP 292, and LNP 334 also showed significant MCP-1 concentrations, LNP 334, which showed high protein expression efficiency, along with LNP 295, is expected to be useful as an mRNA vaccine.

[0466] 9 to 32, LNPs containing compound 16 or compound 17 (LNP 181, 187, 230, 231, 232) showed significantly high protein expression efficiency, but no significant antibody response was observed. On the other hand, LNP 295 containing compound 26 showed a significant antibody response after immunization. LNPs containing compound 36 (LNP 334) showed a significant increase in blood MCP-1 concentration after subcutaneous injection. This indicates that the ionizable lipids with novel structures of the present invention can be tailored to suit the respective applications of therapeutic formulations (LNP 181, 187, 230, 231, 232) that require reduced antibody responses and increased protein expression efficiency, and prophylactic formulations (LNP 295, 334) that require antibody responses.

[0467] Although the present invention has been described in detail above, it is obvious to those skilled in the art that the specific details are merely preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the true scope of the present invention is defined by the claims and their equivalents.

Claims

1. A compound selected from the group consisting of a compound represented by the following chemical formula 1, a stereoisomer thereof, a racemate thereof, and a pharmaceutically acceptable salt thereof: 【Chemistry 1】 In the above Chemical Formula 1, R １ and R ２ are the same or different and each represents hydrogen or a saturated hydrocarbon having 1 to 2 carbon atoms, X １ is O, NH or S, m １ , m ２ and m ３ are the same or different and are integers of 1 to 3, A is hydrogen or a compound represented by the following chemical formula 1-1: 【Chemistry 2】 In the above Chemical Formula 1-1, Z is NR １ R ２ a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, １ and R ２ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; said heterocyclic or aromatic rings are substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and said hydrocarbons may or may not contain ester, ether, amide, carbamate, carbonate or disulfide bonds; X ２ is O or S, Y is CH ２ , NH or O, n １ is an integer from 0 to 3, B is a compound represented by the following chemical formula 1-2, or NR １ 'R ２ ', but the R １ ' and R ２ ' are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R １ ' and R ２ ' are linked to each other to form a heterocycle or aromatic ring having 3 to 8 carbon atoms, 【Transformation 3】 In the above Chemical Formula 1-2, R ３ and R ４ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; E is hydrogen or a compound represented by the following chemical formula 1-3: 【Chemistry 4】 In the above Chemical Formula 1-3, X ３ is O or S, n ２ is an integer from 0 to 3, R ５ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds, and substituted or unsubstituted with thiolane or dithiolane.

2. The compound according to claim 1, wherein the compound represented by Chemical Formula 1 is a compound represented by Chemical Formula 2: 【Transformation 5】 In the above Chemical Formula 2, A １ is hydrogen or a compound represented by the following chemical formula 2-1, 【Transformation 6】 In the above chemical formula 2-1, Z １ is NR ３ R ４ or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, ３ and R ４ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocyclic or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl; n ３ is an integer from 0 to 3, E １ is hydrogen or a compound represented by the following chemical formula 2-2, 【Transformation 7】 In the above Chemical Formula 2-2, R ８ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without an ester, ether, amide, carbamate, carbonate or disulfide bond, and is substituted or unsubstituted with dithiolane; R ６ and R ７ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms.

3. The compound according to claim 1, wherein the compound represented by Chemical Formula 1 is a compound represented by Chemical Formula 3: 【Transformation 8】 In the above Chemical Formula 3, A ２ is hydrogen or a compound represented by the following chemical formula 3-1, 【Chemistry 9】 In the above Chemical Formula 3-1, Z ２ is NR ５ R ６ or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, ５ and R ６ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocyclic or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl; n ４ is an integer from 0 to 3, R ９ and R １０ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; o １ and ２ are the same or different and each is an integer of 2 to 10.

4. The compound according to claim 1, wherein the compound represented by Chemical Formula 1 is a compound represented by Chemical Formula 4: 【Chemistry 10】 In the above Chemical Formula 4, X １ ' is O, NH or S, m ４ and m ５ are the same or different and each represents an integer of 0 to 3, A ３ is a compound represented by the following chemical formula 4-1, 【Chemistry 11】 In the above Chemical Formula 4-1, Z ３ is NR ７ R ８ a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, ７ and R ８ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; said heterocyclic or aromatic rings are substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and said hydrocarbons may or may not contain ester, ether, amide, carbamate, carbonate or disulfide bonds; n ５ is an integer from 0 to 3, B １ is a compound represented by the following chemical formula 4-2; NR ９ R １０ or a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, ９ and R １０ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R ９ and R １０ are linked to each other to form a heterocycle or aromatic ring having 3 to 8 carbon atoms, 【Chemistry 12】 In the above Chemical Formula 4-2, R １２ and R １３ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R １１ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds, and is substituted or unsubstituted with dithiolane.

5. The compound according to claim 2, wherein the compound represented by Chemical Formula 2 is a compound represented by Chemical Formula 5 or Chemical Formula 6: 【Chemistry 13】 In the above Chemical Formula 5, R １４ and R １５ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds. 【Chemistry 14】 In the above Chemical Formula 6, Z ４ is NR １１ R １２ or a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms, １１ and R １２ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; the heterocyclic or aromatic ring is substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl; R １６ and R １７ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds.

6. The compound according to claim 3, wherein the compound represented by Chemical Formula 3 is a compound represented by Chemical Formula 7: 【Chemistry 15】 In the above Chemical Formula 7, R １８ and R １９ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; o １ ' and o ２ ' are the same or different and each is an integer of 3 to 8.

7. The compound according to claim 4, wherein the compound represented by Chemical Formula 4 is a compound represented by Chemical Formula 8 or Chemical Formula 9: 【Chemistry 16】 In the above Chemical Formula 8, m ６ is an integer from 0 to 3, R ２０ and R ２１ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; ２０ and R ２１ are linked to each other to form a heterocycle having 3 to 8 carbon atoms, R ２２ and R ２３ are the same or different and are hydrogen, a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, and wherein the hydrocarbon may or may not contain an ester, ether, amide, carbamate, carbonate, or disulfide bond; R ２４ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester or disulfide bond, and which may or may not be substituted with dithiolane. 【Chemistry 17】 In the above Chemical Formula 9, m ７ and m ８ are the same or different and each represents an integer of 0 to 3, R ２５ is a linear or branched saturated or unsaturated hydrocarbon having 1 to 6 carbon atoms, wherein the hydrocarbon may or may not contain an ester bond or a disulfide bond; R ２６ and R ２７ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms; R ２６ and R ２７ are linked to each other to form a heterocycle or aromatic ring having 3 to 8 carbon atoms, R ２８ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, which may or may not contain an ester or disulfide bond, and which may or may not be substituted with dithiolane.

8. 2. The compound according to claim 1, wherein the heterocycle is selected from the group consisting of substituted or unsubstituted thiophene, furan, pyrazole, pyridine, pyran, oxazine, thiazine, morpholine, pyrrolidine, piperidine, piperazine, pyrazole, pyridine, and dithiolane having 1 to 4 carbon atoms.

9. The compound according to claim 1, wherein the compound represented by Formula 1 is at least one selected from the group consisting of Formulas 10 to 40: 【Chemistry 18-1】 【Chemistry 18-2】 【Chemistry 18-3】 【Chemistry 18-4】 【Chemistry 18-5】

10. A lipid nanoparticle composition comprising the compound of claim 1.

11. The lipid nanoparticle composition of claim 10, wherein the compound is an ionizable lipid or a helper lipid.

12. A lipid nanoparticle composition comprising a compound selected from the group consisting of a compound represented by the following chemical formula 1, a stereoisomer thereof, a racemate thereof, and a pharmaceutically acceptable salt thereof: 【Chemistry 19】 In the above Chemical Formula 1, R １ and R ２ are the same or different and each represents hydrogen or a saturated hydrocarbon having 1 to 2 carbon atoms, X １ is O, NH or S, m １ , m ２ and m ３ are the same or different and are integers of 1 to 3, A is hydrogen or a compound represented by the following chemical formula 1-1: 【Chemistry 20】 In the above Chemical Formula 1-1, Z is NR １ R ２ a heterocyclic or aromatic ring compound having 3 to 8 carbon atoms; or a linear or branched saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, １ and R ２ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms; said heterocyclic or aromatic rings are substituted or unsubstituted with (C1-C4) alkyl or di(C1-C4) alkylamino(C1-2) alkyl, and said hydrocarbons may or may not contain ester, ether, amide, carbamate, carbonate or disulfide bonds; X ２ is O or S, Y is CH ２ , NH or O, n １ is an integer from 0 to 3, B is a compound represented by the following chemical formula 1-2, or NR １ 'R ２ ', but the R １ ' and R ２ ' are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 1 to 6 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; R １ ' and R ２ ' are linked to each other to form a heterocycle or aromatic ring having 3 to 8 carbon atoms, 【Chemistry 21】 In the above Chemical Formula 1-2, R ３ and R ４ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds; E is hydrogen or a compound represented by the following chemical formula 1-3: 【Chemistry 22】 In the above Chemical Formula 1-3, X ３ is O or S, n ２ is an integer from 0 to 3, R ５ is a linear or branched, saturated or unsaturated hydrocarbon having 6 to 22 carbon atoms, with or without ester, ether, amide, carbamate, carbonate or disulfide bonds, and substituted or unsubstituted with thiolane or dithiolane.

13. The lipid nanoparticle composition of claim 12, characterized in that the composition contains one or more selected from the group consisting of helper lipids, structure-retaining lipids, PEG-lipids, and additives.

14. The helper lipids include 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundeca oleoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylethanolamine (POPE), distearoyl-phosphatidyl-ethanolamine (DSPE), dipalmitoyl-phosphatidyl-ethanolamine (DPPE), 1,The lipid nanoparticle composition according to claim 13, characterized in that the lipid nanoparticle is one or more selected from the group consisting of 2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, and lysophosphatidylethanolamine (LPE).

15. The lipid nanoparticle composition of claim 13, wherein the structure-retaining lipid is one or more selected from the group consisting of cholesterol, bile acid derivatives including butyllithocholic acid, cholanic acid derivatives, lithocholic acid derivatives, flavonoids, vitamin A and its derivatives, vitamin E, vitamin K, coenzyme Q10, and β-carotene.

16. The lipid nanoparticle composition of claim 13, wherein the PEG-lipid is one or more selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

17. The lipid nanoparticle composition according to claim 13, wherein the additive is a trehalose derivative represented by the following chemical formula 43: 【Chemistry 23】 In the above Chemical Formula 43, R ３１ and R ３２ are the same or different and are linear or branched, saturated or unsaturated hydrocarbons having 6 to 22 carbon atoms.

18. The composition comprises an ionizable lipid, a helper lipid, a structure-maintaining lipid, a PEG-lipid, and an additive; The composition comprises 20 to 60 mol% of ionizable lipid, 5 to 40 mol% of helper lipid, 25 to 45 mol% of structure-retaining lipid, 1 to 3 mol% of PEG-lipid, and 0 to 25 mol% of additive. The lipid nanoparticle composition of claim 13.

19. The lipid nanoparticle composition according to claim 13, characterized in that the composition contains 20 to 50 mol% of ionizable lipid, 5 to 15 mol% of helper lipid, 25 to 45 mol% of structure-retaining lipid, 1 to 3 mol% of PEG-lipid, and 0 to 25 mol% of additive.

20. The lipid nanoparticle composition according to claim 19, characterized in that the composition comprises a compound of any one of the following formulas 12 to 40 as the ionizable lipid, DSPC, DOPE, or a compound of the following formula 10 or 11 as the helper lipid, cholesterol, a lithocholic acid derivative, or a cholanic acid derivative as the structure-retaining lipid, myristoyl diglyceride (DMG)-PEG as the PEG-lipid, and 6,6'-trehalose dioleate as the additive: 【Chemistry 24-1】 【Chemistry 24-2】 【Chemistry 24-3】 【Chemistry 24-4】 【Chemistry 24-5】

21. 13. The lipid nanoparticle composition of claim 10 or 12, further comprising a therapeutic or prophylactic agent.

22. The lipid nanoparticle composition of claim 21, wherein the therapeutic or prophylactic preparation is a vaccine or a compound capable of inducing an immune response.

23. The lipid nanoparticle composition of claim 21, characterized in that the therapeutic or prophylactic agent is selected from the group consisting of interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof.

24. A composition for drug delivery, comprising the lipid nanoparticle composition of claim 10 or 12; and a therapeutic or prophylactic formulation.

25. An immune enhancing composition comprising the lipid nanoparticle composition of claim 10 or 12.

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