Lipid compounds having a glutaric acid backbone and lipid carriers, nucleic acid lipid nanoparticle compositions and pharmaceutical formulations based thereon

Abstract

The present application belongs to the field of gene drug delivery, and particularly relates to a kind of lipid compound with glutaric acid skeleton and lipid carrier, nucleic acid lipid nanoparticle composition and pharmaceutical preparation based thereon.The compound with the structure of formula (I) of the present application can be prepared into lipid carrier alone or together with other lipid compounds.The lipid carrier has high encapsulation efficiency for nucleic acid drugs, which helps to improve the delivery efficiency of nucleic acid drugs in vivo;Multiple biodegradable functional groups are contained, so that the lipid metabolism is fast, and the biological safety is high.Moreover, the lipid carrier can also deliver nucleic acid drugs to organs that need to be enriched, and has good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of gene drug delivery, specifically relating to a lipid compound, as well as lipid carriers, nucleic acid lipid nanoparticle compositions and pharmaceutical formulations based on the lipid compound. Background Technology

[0002] Gene therapy is a hot research topic in modern biomedicine. For example, nucleic acid drugs can be used to prevent cancer, bacterial and viral infections, and treat diseases with genetic causes. Because nucleic acid drugs are easily degraded and have difficulty entering cells, they need to be encapsulated and delivered to target cells using vectors. Therefore, developing safe and efficient delivery vectors is a prerequisite for the clinical application of gene therapy.

[0003] Lipid nanoparticles (LNPs) are currently a research hotspot in the field of non-viral gene vectors. In 2018, the FDA approved the use of LNPs to deliver patisiran (onpattro) for the treatment of hereditary transthyretin amyloidosis, and since then, research on the delivery of nucleic acid drugs using LNP technology has seen explosive growth. In particular, at the end of 2020, the FDA approved the COVID-19 vaccines from Moderna and BioNTech & Pfizer, both of which utilize LNP technology to deliver mRNA drugs for the prevention of the COVID-19 virus.

[0004] LNPs are typically composed of four types of lipid compounds: cationic lipids, neutral lipids, sterols, and amphiphilic lipids. Among them, cationic lipids have the greatest impact on the performance of LNPs, such as affecting the encapsulation efficiency of nucleic acid drugs, the delivery efficiency of nucleic acid drugs in vivo, or cytotoxicity.

[0005] Therefore, there is a need to develop more novel compounds (such as cationic lipid compounds) to provide more options for delivering gene drugs. Summary of the Invention

[0006] The problem the invention aims to solve

[0007] The present invention aims to provide a series of compounds that can be used alone to prepare lipid carriers or co-prepared with other lipid compounds to prepare lipid carriers, thereby improving the delivery efficiency of nucleic acid drugs in vivo and enabling the delivery of nucleic acid drugs to organs that require enrichment.

[0008] The present invention also provides a lipid carrier comprising the above-described compounds.

[0009] The present invention also provides a nucleic acid lipid nanoparticle composition comprising the above-described compound or the above-described lipid carrier.

[0010] The present invention also provides pharmaceutical formulations comprising the above-described compounds, or the above-described lipid carriers, or the above-described nucleic acid lipid nanoparticle compositions.

[0011] Solution for solving the problem

[0012] In a first aspect, the present invention provides compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs thereof.

[0013]

[0014] in:

[0015] R1 and R2 are each independently C 1-24 Alkyl or C 2-24 alkenyl;

[0016] A1, A2, and A3 are each independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(=O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、-C(R a )OH-、-NR a C(=O)NR a -、-OC(=O)NR a -、-NR a C(=O)O- or -O(C=O)O- or does not exist;

[0017] B1 and B2 are each independently C1-C 12 Alkylene or C2-C 12 The subalkenyl group may not exist;

[0018] B3 is a C1-C6 alkylene group or is absent;

[0019] M1 and M2 are each independently -O- or -NR. a -;

[0020] X is optional. -NR3R4 or -CR connected to B3 a R5R6;

[0021] R3 and R4 are each independently hydrogen or optionally substituted with at least one hydroxyl group or 2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl group. 1-C6 alkyl, or R3 and R4 together with the N they are attached to form one of the following groups that are optionally substituted with at least one C1-C6 alkyl: 3-10 membered heterocyclic alkyl or 5-10 membered heteroaryl;

[0022] R5 and R6 and their connected CR a Together they form one of the following groups optionally substituted with at least one C1-C6 alkyl group: a 3-10-membered nitrogen-containing heterocyclic alkyl group or a 5-10-membered nitrogen-containing heteroaryl group;

[0023] Each R a Each is independently hydrogen or C 1- C6 alkyl;

[0024] The heterocyclic alkyl group and the heteroaryl group each independently have 1 to 3 heteroatoms or heterogroups, each of which is independently N, NH, O, S, S(=O) or S(=O)2; the nitrogen-containing heterocyclic alkyl group and the nitrogen-containing heteroaryl group each independently have 1 to 3 heteroatoms or heterogroups, each of which is independently N, NH, O, S, S(=O) or S(=O)2, and at least one of the heteroatoms or heterogroups is N or NH.

[0025] In a second aspect, the present invention provides specific examples of compounds of formula (I) above, or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs thereof, including (but not limited to):

[0026] Bis(3-((2-Butyloctanoyl)oxy)propyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 1);

[0027] Bis(4-((2-Butyloctanoyl)oxy)butyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 2);

[0028] Bis(5-((2-Butyloctanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 3);

[0029] Bis(6-((2-Butyloctanoyl)oxy)hexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 4);

[0030] Bis(7-((2-Butyloctanoyl)oxy)heptyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 5);

[0031] Bis(8-((2-Butyloctanoyl)oxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 6);

[0032] Bis(3-((2-hexyldecanoyl)oxy)propyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 7);

[0033] Bis(4-((2-hexyldecanoyl)oxy)butyl)3-((4-(dimethylamino)butanoyl)oxy)glutarate (compound 8);

[0034] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 9);

[0035] Bis(6-((2-hexyldecanoyl)oxy)hexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 10);

[0036] Bis(7-((2-hexyldecanoyl)oxy)heptyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 11);

[0037] Bis(8-((2-hexyldecanoyl)oxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 12);

[0038] Bis(3-((2-octyldodecanoyl)oxy)propyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 13);

[0039] Bis(4-((2-octyldodecanoyl)oxy)butyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 14);

[0040] Bis(5-((2-octyldodecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 15);

[0041] Bis(6-((2-octyldodecanoyl)oxy)hexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 16);

[0042] Bis(7-((2-octyldodecanoyl)oxy)heptyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 17);

[0043] Bis(8-((2-octyldodecanoyl)oxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 18);

[0044] Bis(4-((2-Butyloctyl)oxy)-4-oxobutyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 19);

[0045] Bis(5-((2-Butyloctyl)oxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 20);

[0046] Bis(6-((2-Butyloctyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 21);

[0047] Bis(7-((2-Butyloctyl)oxy)-7-oxohepyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 22);

[0048] Bis(8-((2-Butyloctyl)oxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 23);

[0049] Bis(4-((2-hexyldecyl)oxy)-4-oxobutyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 24);

[0050] Bis(5-((2-hexyldecyl)oxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 25);

[0051] Bis(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 26);

[0052] Bis(7-((2-hexyldecyl)oxy)-7-oxohepyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 27);

[0053] Bis(8-((2-hexyldecyl)oxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 28);

[0054] Bis(4-(heptadecano-9-yloxy)-4-oxobutyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 29);

[0055] Bis(5-(heptadecane-9-yloxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 30);

[0056] Bis(6-(heptadecane-9-yloxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 31);

[0057] Bis(7-(heptadecane-9-yloxy)-7-oxohepyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 32);

[0058] Bis(8-(heptadecyl-9-yloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 33);

[0059] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(5-((2-Hexyldecanoyl)oxy)pentyl)3-((4-(Dimethylamino)butyryl)oxy)glutarate (compound 34);

[0060] 1-(5-((2-Butyloctyl)oxy)pentyl)5-(6-((2-Butyloctyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 35);

[0061] 1-(5-((2-Butyloctyl)oxy)pentyl)5-(7-((2-Butyloctyl)oxy)-7-oxohepyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 36);

[0062] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 37);

[0063] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(5-(heptadecano-9-yloxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 38);

[0064] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(6-(heptadecane-9-yloxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 39);

[0065] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 40);

[0066] 1-(6-((2-Butyloctyl)oxy)hexyl)5-(7-((2-Butyloctyl)oxy)-7-oxohepyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 41);

[0067] 1-(6-((2-Butyloctyl)oxy)hexyl)5-(6-((2-Butyloctyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 42);

[0068] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 43);

[0069] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(5-(heptadecane-9-yloxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 44);

[0070] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(6-(heptadecane-9-yloxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 45);

[0071] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 46);

[0072] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 47);

[0073] 1-(5-((2-hexyldecyl)oxy)pentyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 48);

[0074] 1-(5-(heptadecyl-9-yloxy)-5-oxopentyl)5-(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 49);

[0075] 1-(6-(heptadecano-9-yloxy)-6-oxohexyl)5-(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 50);

[0076] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 51);

[0077] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(6-((2-hexyldecyl)oxy)-6-oxohxyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 52);

[0078] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(5-(heptadecane-9-yloxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 53);

[0079] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(6-(heptadecane-9-yloxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 54);

[0080] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(5-(heptadecane-9-yloxy)-5-oxopentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 55);

[0081] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(6-(heptadecane-9-yloxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 56);

[0082] 1-(5-(heptadecyl-9-yloxy)-5-oxopentyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 57);

[0083] 1-(6-(heptadecyl-9-yloxy)-6-oxohexyl)5-(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 58);

[0084] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(8-oxo-8-(undecane-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 59);

[0085] 1-(5-((2-Butyloctanoyl)oxy)pentyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 60);

[0086] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(8-oxo-8-(undecane-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 61);

[0087] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 62);

[0088] 1-(5-((2-hexyldecanoyl)oxy)pentyl)5-(8-oxo-8-(undecano-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 63);

[0089] 1-(5-((2-hexyldecanoyl)oxy)pentyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 64);

[0090] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(8-oxo-8-(undecane-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 65);

[0091] 1-(6-((2-Butyloctyl)oxy)-6-oxohexyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 66);

[0092] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(8-oxo-8-(undecane-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 67);

[0093] 1-(7-((2-Butyloctyl)oxy)-7-oxohepyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 68);

[0094] 1-(6-((2-hexyldecyl)oxy)-6-oxohexyl)5-(8-oxo-8-(undecyl-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 69);

[0095] 1-(6-((2-hexyldecyl)oxy)-6-oxohexyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 70);

[0096] 1-(6-(heptadecane-9-yloxy)-6-oxohexyl)5-(8-oxo-8-(undecane-3-yloxy)octyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 71);

[0097] 1-(6-(heptadecyl-9-yloxy)-6-oxohexyl)5-(8-(nonyloxy)-8-oxooctyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 72);

[0098] 1-(6-((2-Butyloctanoyl)oxy)hexyl)5-((9E,12E)-octadecane-9,12-dien-1-yl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 73);

[0099] 1-(6-((2-hexyldecanoyl)oxy)hexyl)5-((9E,12E)-octadecano-9,12-dien-1-yl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 74);

[0100] 1-(6-((2-hexyldecyl)oxy)-6-oxohexyl)5-((9E,12E)-octadecane-9,12-dien-1-yl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 75);

[0101] 1-(6-(heptadecane-9-yloxy)-6-oxohexyl)5-((9E,12E)-octadecane-9,12-dien-1-yl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 76);

[0102] Di((9E,12E)-octadecano-9,12-dien-1-yl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 77);

[0103] Bis(4-(((9E,12E)-heptadecane-9,12-dienoyl)oxy)butyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 78);

[0104] 7,29-Dibutyl-8,16,20,28-tetraoxo-9,15,21,27-tetraaza-triapentadecan-18-yl 4-(dimethylamino)butyrate (compound 79);

[0105] 7,31-Dibutyl-8,17,21,30-tetraoxo-9,16,22,29-tetraaza-hepta-heptadecane-19-yl 4-(dimethylamino)butyrate (compound 80);

[0106] 9,31-Dihexyl-10,18,22,30-tetraoxo-11,17,23,29-tetraaza-nonadecan-20-yl 4-(dimethylamino)butyrate (compound 81);

[0107] 11,33-Dioctyl-12,20,24,32-Tetraoxo-13,19,25,31-Tetraaza-tetratriatriane-22-yl 4-(dimethylamino)butyrate (Compound 82);

[0108] 7,31-Dibutyl-10,17,21,28-tetraoxo-9,16,22,29-tetraaza-hepta-heptadecane-19-yl 4-(dimethylamino)butyrate (compound 83);

[0109] 7,33-Dibutyl-10,18,22,30-tetraoxo-9,17,23,31-tetraaza-nonadecan-20-yl 4-(dimethylamino)butyrate (compound 84);

[0110] 9,33-Dihexyl-12,19,23,30-tetraoxo-11,18,24,31-tetraaza-tetradecane-21-yl 4-(dimethylamino)butyrate (compound 85);

[0111] 9,29-Dioctyl-11,17,21,27-Tetraoxo-10,16,22,28-Tetraaza-hepta-heptadecane-19-yl 4-(dimethylamino)butyrate (Compound 86);

[0112] 9-Hexyl-10,18,22-trioxo-26,27-dithia-11,17,23-triazatetran-20-yl-4-(dimethylamino)butyrate (compound 87);

[0113] 9,31-Dioctyl-11,18,22,29-tetraoxo-10,17,23,30-tetraaza-nonadecan-20-yl 4-(dimethylamino)butyrate (compound 88);

[0114] N 1 N 5 -Bis(5-(2-Butyloctanoylamino)pentyl)-3-(4-(dimethylamino)butyrylamino)glutaramide (compound 89);

[0115] N 1 N 5 -Bis(6-(2-Butyloctanoylamino)hexyl)-3-(4-(dimethylamino)butanoylamino)glutaramide (compound 90);

[0116] 3-(4-(dimethylamino)butyrylamino)-N 1 N 5 -Bis(5-(2-hexyldecanoylamino)pentyl)glutaramide (compound 91);

[0117] 3-(4-(dimethylamino)butyrylamino)-N 1 N 5 -Bis(5-(2-octyldodecanoylamino)pentyl)glutaramide (compound 92);

[0118] N 1 N 5 -Bis(6-((2-Butyloctyl)amino)-6-oxohexyl)-3-(4-(dimethylamino)butyrylamino)glutaramide (compound 93);

[0119] N 1 N 5 -Bis(7-((2-Butyloctyl)amino)-7-oxohepyl)-3-(4-(dimethylamino)butyrylamino)glutaramide (compound 94);

[0120] 3-(4-(dimethylamino)butyrylamino)-N 1 N 5 -Bis(6-((2-hexyldecyl)amino)-6-oxohexyl)glutaramide (compound 95);

[0121] 3-(4-(dimethylamino)butyrylamino)-N 1 N 5 - bis(5-(heptadecane-9-ylamino)-5-oxopentyl)glutaramide (compound 96);

[0122] 3-(4-(dimethylamino)butyrylamino)-N 1 N 5- bis(6-(heptadecane-9-ylamino)-6-oxohexyl)pentanediamide (compound 97);

[0123] N 1 N 3 -bis(5-(2-Butyloctanoylamino)pentyl)-N 2 -(3-(dimethylamino)propyl)propane-1,2,3-tricarboxamide (compound 98);

[0124] N 1 N 3 -bis(6-(2-Butyloctanoylamino)hexyl)-N 2 -(3-(dimethylamino)propyl)propane-1,2,3-tricarboxamide (compound 99);

[0125] N 2 -(3-(dimethylamino)propyl)-N 1 N 3 -Bis(5-(2-hexyldecanoylamino)pentyl)propane-1,2,3-tricarboxamide (compound 100);

[0126] N 2 -(3-(dimethylamino)propyl)-N 1 N 3 - bis(5-(2-octyldodecanoylamino)pentyl)propane-1,2,3-tricarboxamide (compound 101);

[0127] N 1 N 3 -bis(6-((2-butyloctyl)amino)-6-oxohexyl)-N 2 -(3-(dimethylamino)propyl)propane-1,2,3-tricarboxamide (compound 102);

[0128] N 1 N 3 -bis(7-((2-butyloctyl)amino)-7-oxohepyl)-N 2 -(3-(dimethylamino)propyl)propane-1,2,3-tricarboxamide (compound 103);

[0129] N 2 -(3-(dimethylamino)propyl)-N 1 N 3 -bis(6-((2-hexyldecyl)amino)-6-oxohexyl)propane-1,2,3-tricarboxamide (compound 104);

[0130] N 2 -(3-(dimethylamino)propyl)-N 1 N3 -bis(5-(heptadecane-9-ylamino)-5-oxopentyl)propane-1,2,3-tricarboxamide (compound 105);

[0131] N 2 -(3-(dimethylamino)propyl)-N 1 N 3 -bis(6-(heptadecane-9-ylamino)-6-oxohexyl)propane-1,2,3-tricarboxamide (compound 106);

[0132] ((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))bis(pentane-5,1-diyl)bis(2-butyloctanoate) (compound 107);

[0133] ((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))bis(hexane-6,1-diyl)bis(2-butyloctanoate) (compound 108);

[0134] ((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))bis(pentane-5,1-diyl)bis(2-hexyldecanoate) (compound 109);

[0135] ((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))bis(pentane-5,1-diyl)bis(2-octyldodecanoate) (compound 110);

[0136] Bis(2-butyloctyl)6,6'-((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))dihexanoate (compound 111);

[0137] Bis(2-butyloctyl)7,7'-((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))diheptanoate (compound 112);

[0138] Bis(2-hexyldecyl)6,6'-((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))dihexanoate (compound 113);

[0139] Di(heptadecyl-9-yl)5,5'-((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))divalerate (compound 114);

[0140] Di(heptadecyl-9-yl)6,6'-((3-(4-(dimethylamino)butyrylamino)glutaryl)bis(azanediyl))dihexanoate (compound 115);

[0141] Bis(5-(2-Butyloctanoylamino)pentyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 116);

[0142] Bis(6-(2-Butyloctanoylamino)hexyl)3-(4-(dimethylamino)butanoylamino)glutarate (compound 117);

[0143] Bis(5-(2-hexyldecanoylamino)pentyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 118);

[0144] Bis(5-(2-octyldodecanoylamino)pentyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 119);

[0145] Bis(6-((2-butyloctyl)amino)-6-oxohexyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 120);

[0146] Bis(7-((2-butyloctyl)amino)-7-oxohepyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 121);

[0147] Bis(6-((2-hexyldecyl)amino)-6-oxohexyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 122);

[0148] Bis(5-(heptadecano-9-ylamino)-5-oxopentyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 123);

[0149] Bis(6-(heptadecano-9-ylamino)-6-oxohexyl)3-(4-(dimethylamino)butyrylamino)glutarate (compound 124);

[0150] Bis(5-((2-Butyloctanoyl)oxy)pentyl)3-((4-(Ethyl(methyl)amino)butyryl)oxy)glutarate (compound 125);

[0151] Bis(6-((2-Butyloctanoyl)oxy)hexyl)3-((4-(Ethyl(methyl)amino)butyryl)oxy)glutarate (compound 126);

[0152] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(ethyl(methyl)amino)butyryl)oxy)glutarate (compound 127);

[0153] Bis(6-((2-Butyloctyl)oxy)-6-oxohexyl)3-((4-(ethyl(methyl)amino)butyryl)oxy)glutarate (compound 128);

[0154] Bis(6-((2-hexyldecyl)oxy)-6-oxohexyl)3-((4-(ethyl(methyl)amino)butyryl)oxy)glutarate (compound 129);

[0155] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-((5-(dimethylamino)pentanoyl)oxy)glutarate (compound 130);

[0156] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(diethylamino)butyryl)oxy)glutarate (compound 131);

[0157] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-(((2-(dimethylamino)ethoxy)carbonyl)amino)glutarate (compound 132);

[0158] Bis(5-((2-hexyldecanoyl)oxy)pentyl)3-((4-(1H-imidazol-1-yl)butyryl)oxy)glutarate (compound 133);

[0159] 1-(5-((2-hexyldecanoyl)oxy)pentyl)5-(2-(tetradecyldithioyl)ethyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 134);

[0160] Bis(5-((((2-hexyldecyl)oxy)carbonyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 135);

[0161] bis(5-(((heptadecyl-9-yloxy)carbonyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 136); and

[0162] 1-(5-((((2-Butyloctyl)oxy)carbonyl)oxy)pentyl)5-(5-(((heptadecane-9-yloxy)carbonyl)oxy)pentyl)3-((4-(dimethylamino)butyryl)oxy)glutarate (compound 137).

[0163] Thirdly, the present invention provides a lipid carrier comprising the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug.

[0164] Fourthly, the present invention provides a nucleic acid lipid nanoparticle composition comprising the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug, or the above-mentioned lipid carrier, and a nucleic acid drug.

[0165] Fifthly, the present invention provides a pharmaceutical formulation comprising the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, or the above-mentioned lipid carrier, or the above-mentioned nucleic acid lipid nanoparticle composition, and a pharmaceutically acceptable excipient, carrier and diluent.

[0166] The effects of the invention

[0167] This invention provides a series of novel compounds of formula (I), which, as cationic lipids, can be used alone to prepare lipid carriers or co-prepared with other lipid compounds. They exhibit controllable particle size, uniform distribution, monodispersity, and high encapsulation efficiency for negatively charged drugs. These cationic lipids demonstrate high biosafety; they contain multiple degradable functional groups, enabling them to facilitate greater release and expression of nucleic acids in vivo; and they also promote faster lipid metabolism. Furthermore, these lipid carriers can deliver nucleic acid drugs to organs requiring enrichment.

[0168] Furthermore, the compound has a simple synthetic route, uses inexpensive and readily available raw materials, and has high market potential. Attached Figure Description

[0169] Figure 1 This is a mouse in vivo fluorescence imaging image of LNP@luci-mRNA formed from compound 4 of the present invention.

[0170] Figure 2 This is a mouse in vivo fluorescence imaging image of LNP@luci-mRNA formed from compound 21 of the present invention.

[0171] Figure 3 This is an in vitro fluorescence imaging image of LNP@luci-mRNA formed from compound 21 of the present invention in mice.

[0172] Figure 4 This is a mouse in vivo fluorescence imaging image of LNP@luci-mRNA formed from compound 45 of the present invention.

[0173] Figure 5 This is an in vitro fluorescence imaging image of LNP@luci-mRNA formed from compound 45 of the present invention in mice. Detailed Implementation

[0174] Before further describing the invention, it should be understood that the invention is not limited to the specific embodiments described herein; it should also be understood that the terminology used herein is for description only and not for limiting specific embodiments.

[0175] [Terminology Definition]

[0176] Unless otherwise stated, the following terms have the following meanings:

[0177] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention that is substantially non-toxic to organisms. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by the reaction of the compounds of this invention with pharmaceutically acceptable inorganic / organic acids or inorganic / organic bases; such salts are also known as acid addition salts or base addition salts. Common inorganic acids include (but are not limited to) hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.; common organic acids include (but are not limited to) trifluoroacetic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, oxalic acid, formic acid, acetic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.; common inorganic bases include (but are not limited to) sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, etc.; and common organic bases include (but are not limited to) diethylamine, triethylamine, ethylaminobutanol, etc.

[0178] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a perpendicular asymmetric plane due to having at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.), thereby enabling the rotation of plane-polarized light. Since the compounds of this invention contain asymmetric centers and other chemical structures that may lead to stereoisomerism, this invention also includes these stereoisomers and mixtures thereof. Because the compounds of this invention and their salts include asymmetric carbon atoms, they can exist as single stereoisomers, racemates, or mixtures of enantiomers and diastereomers. Typically, these compounds can be prepared as racemic mixtures. However, if desired, such compounds can be prepared or isolated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched with single stereoisomers (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85%, or ≥80%). The single stereoisomer of a compound is synthesized from an optically active starting material containing the desired chiral center, or obtained by preparing a mixture of enantiomers followed by separation or resolution, for example, by converting it into a mixture of diastereomers followed by separation or recrystallization, chromatographic treatment, using chiral resolving reagents, or by direct separation of the enantiomers on a chiral chromatographic column. Starting compounds with specific stereochemistry are commercially available or can be prepared according to the methods described below and then resolved by methods well known in the art.

[0179] The term "tautomer" (or "tautomer form") refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (or proton transfer tautomers) include (but are not limited to) interconversions via proton transfer, such as keto-enol isomerization, imine-enamine isomerization, amide-imine alcohol isomerization, etc. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0180] The term "solvent" refers to a substance formed by the combination of a compound of the present invention or a pharmaceutically acceptable salt thereof with at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates, ethanol compounds, acetone compounds, etc.

[0181] The term "chelate" refers to a complex with a cyclic structure, obtained through the chelation of two or more ligands with the same metal ion to form a chelate ring.

[0182] The term "non-covalent complex" refers to a complex formed through the interaction of a compound with another molecule, where no covalent bond is formed between the two molecules. Complexation can occur, for example, through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also known as ionic bonding).

[0183] The term "prodrug" refers to a derived compound that, when administered to a patient, can directly or indirectly provide the compounds of the present invention. Particularly preferred derived compounds or prodrugs are those that, when administered to a patient, can improve the bioavailability of the compounds of the present invention (e.g., facilitate absorption into the bloodstream) or promote the delivery of the parent compound to its site of action (e.g., the lymphatic system). Unless otherwise stated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

[0184] The term "independently" means that at least two groups (or ring systems) in a structure with the same or similar value ranges can have the same or different meanings under specific circumstances. For example, if substituent X and substituent Y are independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl.

[0185] The term “optional” or “optionally” means that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0186] The terms “contain” and “include” are used in their open, non-restrictive sense.

[0187] The term "alkyl" refers to a monovalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, without unsaturation, and linked to other segments by a single bond, including (but not limited to) methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, and tert-butyl. For example, "C1-C..." 24 "Alkyl" refers to an alkyl group containing 1 to 24 carbon atoms. Specifically, the term "branched C" refers to an alkyl group... 10 -C 15 "Alkyl" refers to a branched alkyl group containing 10 to 15 carbon atoms, including (but not limited to) 1-butylhept-1-yl and 2-butyloct-1-yl.

[0188] The term "alkylene" refers to a divalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, without saturation, and connected to other segments by two single bonds, including (but not limited to) methylene, 1,2-ethylene, 1,3-propylene, and 1,4-butylene. For example, "C1-C..." 12 "Alkylene" refers to an alkylene group containing 1 to 12 carbon atoms.

[0189] The term "alkenyl" refers to a monovalent, straight-chain or branched aliphatic group composed only of carbon and hydrogen atoms, containing at least one double bond, and connected to other segments by a single bond, including (but not limited to) vinyl, propenyl, allyl, and other groups. For example, "C2-C..." 24 "Alkenyl" refers to an alkenyl group containing 2 to 24 carbon atoms.

[0190] The term "alkenyl" refers to a divalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, containing at least one double bond, and connected to other segments via two single bonds, including (but not limited to) other segments. For example, "C2-C" 12 "Alkenyl" refers to alkenyl groups containing 2 to 12 carbon atoms.

[0191] The term "cycloalkyl" refers to a monovalent monocyclic or polycyclic (e.g., fused, bridged, or spirocyclic) aliphatic group consisting only of carbon and hydrogen atoms and linked to other segments by a single bond, including (but not limited to) cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. For example, "C3-C..." 24 "Alkyl" refers to a cycloalkyl group containing 3 to 24 cyclic atoms.

[0192] The term "heterocyclic alkyl" refers to a monovalent monocyclic or polycyclic (e.g., fused, bridged, or spirocyclic) aliphatic group consisting of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are each independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by a single bond, including (but not limited to) pyrrolid-1-yl, piperidin-1-yl, and 4-methylpiperazin-1-yl. For example, "3-24 membered heterocyclic alkyl" refers to a heterocyclic alkyl containing 3 to 24 cyclic atoms (or heterogroups). The term "nitrogen-containing heterocyclic alkyl" refers to a heterocyclic alkyl with at least one ring atom (or ring group) being an N atom (or containing an N group, e.g., an N→O group).

[0193] The term "aryl" refers to a monovalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group consisting only of carbon and hydrogen atoms and linked to other segments by a single bond, including (but not limited to) phenyl, naphthyl, anthracene, and phenanthrene groups. For example, "C6-C..." 10 "Aryl" refers to an aryl group containing 6 to 10 cyclic atoms.

[0194] The term "heteroaryl" refers to a monovalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group composed of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by a single bond, including (but not limited to) pyrazolyl (e.g., 1H-imidazol-1-yl), oxazolyl (e.g., oxazol-2-yl), and thiazolyl (e.g., thiazolyl-4-yl). For example, "5-10-membered heteroaryl" refers to a heteroaryl containing 5 to 10 cyclic atoms (or heterogroups). The term "nitrogen-containing heteroaryl" refers to a heteroaryl with at least one cyclic atom (or cyclic group) being an N atom (or containing an N-group, e.g., an NH-group).

[0195] The term "hydroxyl group" refers to the -OH group.

[0196] The term "2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl" refers to Group.

[0197] [General Formula Compound]

[0198] This invention provides compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs thereof.

[0199]

[0200] in:

[0201] R1 and R2 are each independently C1-24 Alkyl or C 2-24 alkenyl;

[0202] A1, A2, and A3 are each independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(=O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、-C(R a )OH-、-NR a C(=O)NR a -、-OC(=O)NR a -、-NR a C(=O)O- or -O(C=O)O- or does not exist;

[0203] B1 and B2 are each independently C1-C 12 Alkylene or C2-C 12 The subalkenyl group may not exist;

[0204] B3 is a C1-C6 alkylene group or is absent;

[0205] M1 and M2 are each independently -O- or -NR. a -;

[0206] X is optional. -NR3R4 or -CR connected to B3 a R5R6;

[0207] R3 and R4 are each independently hydrogen or optionally substituted with at least one hydroxyl group or 2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl group. 1- C6 alkyl, or R3 and R4 together with the N they are attached to form one of the following groups that are optionally substituted with at least one C1-C6 alkyl: 3-10 membered heterocyclic alkyl or 5-10 membered heteroaryl;

[0208] R5 and R6 and their connected CR a Together they form one of the following groups optionally substituted with at least one C1-C6 alkyl group: a 3-10-membered nitrogen-containing heterocyclic alkyl group or a 5-10-membered nitrogen-containing heteroaryl group;

[0209] Each R a Each is independently hydrogen or C 1- C6 alkyl;

[0210] The heterocyclic alkyl group and the heteroaryl group each independently have 1 to 3 heteroatoms or heterogroups, each of which is independently N, NH, O, S, S(=O) or S(=O)2; the nitrogen-containing heterocyclic alkyl group and the nitrogen-containing heteroaryl group each independently have 1 to 3 heteroatoms or heterogroups, each of which is independently N, NH, O, S, S(=O) or S(=O)2, and at least one of the heteroatoms or heterogroups is N or NH.

[0211] In some embodiments, A1, A2, and A3 in the compound of formula (I) are each independently -O(C=O)-, -(C=O)O-, -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、-NR a C(=O)NR a -、-OC(=O)NR a -、-NR a C(=O)O- or -O(C=O)O- or does not exist, where R a As defined in equation (I).

[0212] In some embodiments, A1, A2, and A3 in the compound of formula (I) are each independently -O(C=O)-, -(C=O)O-, -SS-, and -NR-. a C(=O)-、-C(=O)NR a -、-OC(=O)NR a -、-NR a C(=O)O- or -O(C=O)O- or does not exist, where R a As defined in equation (I).

[0213] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 4-24 Alkyl or C 4-24 Alkenyl group.

[0214] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 4-24 Straight-chain alkyl or C 4-24 Straight-chain alkenyl groups.

[0215] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 9-24 Straight-chain alkyl or C 9-24 Straight-chain alkenyl groups.

[0216] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 9-18Straight-chain alkyl or C 9-18 Straight-chain alkenyl groups.

[0217] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 4-24 Branched alkyl or C 4-24 Branched alkenyl groups.

[0218] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 11-24 Branched alkyl or C 11-24 Branched alkenyl groups.

[0219] In some embodiments, R1 and R2 in the compound of formula (I) are each independently C 11 -19 branched alkyl groups or C 11 -19 branched alkenyl groups.

[0220] In some embodiments, R1 and R2 in the compound of formula (I) are each independently non-1-yl, dec-1-yl, undec-1-yl, dodec-1-yl, tridec-1-yl, tetradec-1-yl, pentadec-1-yl, 2-butylhexyl-1-yl, 2-butyloctyl-1-yl, 2-butyldec-1-yl, 2-hexylhexyl-1-yl, 2-hexyloctyl-1-yl, 2-hexyldec-1-yl, 2-octyloctyl-1-yl, 2-octyl 2-decyldecyl-1-yl, 2-octyldodec-1-yl, undecyl-5-yl, undecyl-6-yl, tridecyl-7-yl, pentadecyl-7-yl, heptadecanyl-9-yl, nonadecanyl-9-yl, non-2-en-1-yl, decyl-4-en-1-yl, hexadec-8,11-dien-1-yl, octadec-9,12-dien-1-yl or 2-(decyl-4-en-1-yl)dodecyl-6-en-1-yl.

[0221] In some embodiments, R1 and R2 in the compound of formula (I) are each independently non-1-yl, dec-1-yl, tridecan-1-yl, tetradecan-1-yl, 2-butylhex-1-yl, 2-butyloct-1-yl, 2-butyldecan-1-yl, 2-hexylhex-1-yl, 2-hexyloct-1-yl, 2-hexyldecan-1-yl, undecan-5-yl, tridecan-7-yl, pentadecan-7-yl, heptadecan-9-yl, nonadecan-9-yl, non-2-en-1-yl, decan-4-en-1-yl, hexadec-8,11-dien-1-yl, octadec-9,12-dien-1-yl or 2-(decan-4-en-1-yl)dodecan-6-en-1-yl.

[0222] In some embodiments, R1 and R2 in the compound of formula (I) are each independently non-1-yl, tetradec-1-yl, 2-butyloct-1-yl, 2-hexyldec-1-yl, undec-5-yl, pentadec-7-yl, heptadecan-9-yl, nonadecan-9-yl, hexadec-8,11-dien-1-yl or octadec-9,12-dien-1-yl.

[0223] In some embodiments, B1 and B2 in the compound of formula (I) are each independently C1-C 10 Alkylene or C2-C 10 The subalkenyl group may not exist.

[0224] In some embodiments, B1 and B2 in the compound of formula (I) are each independently C2-C8 alkylene or C2-C8 alkenylene or absent.

[0225] In some embodiments, B1 and B2 in the compound of formula (I) are each independently C2-C8 alkylene groups or are absent.

[0226] In some embodiments, B3 in the compound of formula (I) is a C1-C4 alkylene group or is absent.

[0227] In some embodiments, B3 in the compound of formula (I) is a C2-C4 alkylene group.

[0228] In some embodiments, M1 and M2 in the compound of formula (I) are both -O- or -NR. a -, where R a As defined in equation (I).

[0229] In some embodiments, M1 and M2 in the compound of formula (I) are both -O-.

[0230] In some embodiments, M1 and M2 in the compound of formula (I) are both -NR. a -, where R a As defined in equation (I).

[0231] In some embodiments, X in the compound of formula (I) is -NR3R4, wherein R3 and R4 are each independently hydrogen or optionally substituted with at least one hydroxyl group or 2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl C 1- C4 alkyl, or R3 and R4 together with the N attached thereto form one of the following groups optionally substituted with at least one C1-C6 alkyl: 3-6 membered heterocyclic alkyl or 5-6 membered heteroaryl.

[0232] In some embodiments, X in the compound of formula (I) is -NR3R4, wherein R3 and R4 are each independently hydrogen or optionally substituted with at least one hydroxyl group or 2-(methylamino)-3,4-dioxocyclobut-1-en-1-yl of one of the following groups: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl; or, R3 and R4 together with the N attached thereto form one of the following groups optionally substituted with at least one methyl, ethyl, n-propyl, or isopropyl of one of the following groups: acridine-1-yl, acridine-1-yl, pyrrolidine-1-yl, piperidin-1-yl, pyrrolidinyl, or 1H-imidazol-1-yl.

[0233] In some embodiments, X in the compound of formula (I) is any one of the following groups:

[0234]

[0235] In some embodiments, X in the compound of formula (I) is any one of the following groups:

[0236]

[0237] In some embodiments, X in the compound of formula (I) is any one of the following groups:

[0238]

[0239] In some implementations, each R in the compound of formula (I) a Each is independently hydrogen or C 1- C4 alkyl.

[0240] In some implementations, each R in the compound of formula (I) a Each is independently hydrogen.

[0241] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-1).

[0242]

[0243] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0244] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-2).

[0245]

[0246] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0247] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-3).

[0248]

[0249] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0250] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-4).

[0251]

[0252] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0253] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-5).

[0254]

[0255] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0256] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-6).

[0257]

[0258] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0259] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-7).

[0260]

[0261] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen. In some embodiments, the compound of formula (I) is a compound as shown in formula (I-8).

[0262]

[0263] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0264] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-9).

[0265]

[0266] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0267] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-10).

[0268]

[0269] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0270] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-11).

[0271]

[0272] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0273] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-12).

[0274]

[0275] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen. In some embodiments, the compound of formula (I) is a compound as shown in formula (I-13).

[0276]

[0277] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0278] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-14).

[0279]

[0280] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0281] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-15).

[0282]

[0283] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0284] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-16).

[0285]

[0286] Where: R1, R2, B1, B2, B3, X and R a As defined in equation (I); preferably, R a It is hydrogen.

[0287] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-17).

[0288]

[0289] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0290] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-18).

[0291]

[0292] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0293] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-19).

[0294]

[0295] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0296] In some embodiments, the compound of formula (I) is a compound as shown in formula (I-20).

[0297]

[0298] Where R1, R2, B1, B2, B3 and X are as defined in equation (I).

[0299] In some embodiments, X in compounds of formulas (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), (I-7), (I-8), (I-9), (I-10), (I-11), (I-12), (I-13), (I-14), (I-15), (I-16), (I-17), (I-18), (I-19), and (I-20) is independently... Preferred

[0300] [Specific compound]

[0301] This invention provides a series of specific compounds, including (but not limited to) the following compounds.

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316] [Lipid carrier]

[0317] This invention provides a lipid carrier comprising any of the aforementioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs. This type of lipid carrier exhibits high encapsulation efficiency for nucleic acid drugs, significantly improving the in vivo delivery efficiency of nucleic acid drugs.

[0318] In some embodiments, the lipid carrier comprises a first lipid compound and a second lipid compound, wherein the first lipid compound comprises any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs and optionally cationic lipids, and the second lipid compound comprises one or more of anionic lipids, neutral lipids, sterols and amphiphilic lipids.

[0319] In some specific implementations, the first lipid compound is any of the above-mentioned compounds or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug.

[0320] In some other specific embodiments, the first lipid compound is any of the above-mentioned compounds or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or a combination of a prodrug and a cationic lipid.

[0321] In some specific implementations, the second lipid compound is a combination of neutral lipids, sterols, and amphiphilic lipids.

[0322] In other specific embodiments, the second lipid compound is a combination of anionic lipids, neutral lipids, sterols, and amphiphilic lipids.

[0323] In some specific embodiments, the above-mentioned cationic lipids include (but are not limited to) one or more combinations of DLinDMA, DODMA, DLin-MC2-MPZ, DLin-KC2-DMA, DOTAP, C12-200, DC-Chol and DOTMA, with DLin-KC2-DMA and DOTAP being preferred.

[0324] In some specific embodiments, the above-mentioned anionic lipids include (but are not limited to) one or more combinations of phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, DOPG, DOPS and myristoyl phosphatidylglycerol, preferably DOPG and DOPS.

[0325] In some specific embodiments, the aforementioned neutral lipids include (but are not limited to) at least one of DOPE, DSPC, DPPC, DOPC, DPPG, POPC, POPE, DPPE, DMPE, DSPE, and SOPE, or lipids modified with anionic or cationic modifying groups, preferably DSPC. The anionic or cationic modifying groups are not limited.

[0326] In some specific embodiments, the aforementioned amphiphilic lipids include (but are not limited to) one or more combinations of PEG-DMG, PEG-c-DMG, PEG-C14, PEG-c-DMA, PEG-DSPE, PEG-PE, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, Tween-20, Tween-80, PEG-DPG, PEG-s-DMG, DAA, PEG-c-DOMG, and GalNAc-PEG-DSG, preferably PEG-DMG and Tween-80.

[0327] In some specific implementations, in the lipid carrier, the molar ratio of the first lipid compound, anionic lipid, neutral lipid, sterol, and amphiphilic lipid is (20–65):(0–20):(5–25):(25–55):(0.3–15); exemplaryly, the molar ratio can be 20:20:5:50:5, 30:5:25:30:10, 20:5:5:55:15, 65:0:9.7:25:0.3, etc.; wherein, in the first lipid compound, the molar ratio of any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs and cationic lipids is (1–10):(0–10); exemplaryly, the molar ratio can be 1:1, 1:2, 1:5, 1:7.5, 1:10, 2:1, 5:1, 7.5:1, 10:1, etc.

[0328] In some more specific embodiments, in the lipid carrier, the molar ratio of the first lipid compound, anionic lipid, neutral lipid, sterol and amphiphilic lipid is (20-55):(0-13):(5-25):(25-51.5):(0.5-15); wherein, in the first lipid compound, the molar ratio of any of the above compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs and cationic lipids is (3-4):(0-5).

[0329] [Nucleic Acid Nanoparticle Composition]

[0330] The present invention provides a nucleic acid nanoparticle composition comprising any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs or the above-mentioned lipid carriers, as well as a nucleic acid drug.

[0331] In some implementations, the aforementioned nucleic acid drugs include (but are not limited to) one or more combinations of DNA, siRNA, mRNA, dsRNA, antisense nucleic acid, antisense oligonucleotide, microRNA, antisense microRNA, antagomir, microRNA inhibitors, microRNA activators, and immunostimulatory nucleic acids.

[0332] In some specific implementation schemes, the mass ratio of the above-mentioned nucleic acid drug to any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs is 1:(3-40).

[0333] In some other specific implementations, the mass ratio of the above-mentioned nucleic acid drug to the above-mentioned lipid carrier is 1:(3-40).

[0334] For example, the above mass ratio can be 1:3, 1:5, 1:10, 1:15, 1:20, 1:30, etc.

[0335] [Pharmaceutical Preparations]

[0336] The present invention provides a pharmaceutical formulation comprising any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs, or the above-mentioned lipid carriers, or the above-mentioned nucleic acid lipid nanoparticle compositions, as well as pharmaceutically acceptable excipients, carriers and diluents.

[0337] In some embodiments, the particle size of the above-mentioned pharmaceutical formulation is 30 to 500 nm; for example, the particle size can be 30 nm, 50 nm, 100 nm, 150 nm, 250 nm, 350 nm, 500 nm, etc.

[0338] In some specific implementation schemes, the encapsulation rate of the nucleic acid drug in the above-mentioned drug formulation is greater than 50%; for example, the encapsulation rate can be 55%, 60%, 65%, 70%, 75%, 79%, 80%, 85%, 89%, 90%, 93%, 95%, etc.

[0339] [Preparation Method]

[0340] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0341] In this invention, the "equivalent (eq)" ratio refers to the molar ratio of the solvent or the drug.

[0342] In this invention, "appropriate amount" means that the amount of solvent or reagent added can be adjusted within a large range and has little impact on the synthesis result, and no specific limitation is required.

[0343] In the following examples, all solvents and reagents used were of analytical or chemical purity; all solvents were redistilled before use; and all anhydrous solvents were processed according to standard or literature methods.

[0344] Example 1: Synthesis of Compound 3

[0345] 1,5-Pentanediol (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-butyloctanoic acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0346] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0347] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0348] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 3.

[0349] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.42-4.16(m,8H),2.43-2.33(m, 8H),2.34(s,6H),1.98-1.76(m,4H),1.68-1.20(m,44H),0.92-0.88(m,12H).

[0350] Example 2: Synthesis of Compound 4

[0351] 1,6-Hexanediol (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-butyloctanoic acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0352] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0353] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0354] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 4.

[0355] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.42-4.16(m,8H),2.43-2.33(m, 8H),2.34(s,6H),1.98-1.76(m,4H),1.68-1.20(m,48H),0.92-0.88(m,12H).

[0356] Example 3: Synthesis of Compound 9

[0357] 1,5-Pentanediol (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-hexyldecanoic acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0358] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0359] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0360] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 9.

[0361] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.42-4.16(m,8H),2.43-2.33(m, 8H),2.34(s,6H),1.98-1.76(m,4H),1.68-1.20(m,60H),0.92-0.88(m,12H).

[0362] Example 4: Synthesis of Compound 15

[0363] 1,5-Pentanediol (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-octyldodecanoic acid (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 1.

[0364] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0365] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0366] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 15.

[0367] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.42-4.16(m,8H),2.43-2.33(m, 8H),2.34(s,6H),1.98-1.76(m,4H),1.68-1.20(m,76H),0.92-0.88(m,12H).

[0368] Example 5: Synthesis of Compound 21

[0369] 6-Hydroxyhexanoic acid (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-butyloctanol (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an empty container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0370] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0371] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0372] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 21.

[0373] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.56-4.11(m,8H),2.43-2.33(m, 12H),2.34(s,6H),1.94-1.81(m,4H),1.68-1.20(m,44H),0.92-0.88(m,12H).

[0374] Example 6: Synthesis of Compound 22

[0375] 7-Hydroxyheptanoic acid (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-butyloctanol (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0376] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0377] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0378] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 22.

[0379] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.56-4.11(m,8H),2.43-2.33(m, 12H),2.34(s,6H),1.94-1.81(m,4H),1.68-1.20(m,48H),0.92-0.88(m,12H).

[0380] Example 7: Synthesis of Compound 26

[0381] 6-Hydroxyhexanoic acid (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 2-hexyldecyl alcohol (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0382] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0383] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0384] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 26.

[0385] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.56-4.11(m,8H),2.43-2.33(m, 12H),2.34(s,6H),1.94-1.81(m,4H),1.68-1.20(m,60H),0.92-0.88(m,12H).

[0386] Example 8: Synthesis of Compound 30

[0387] 5-Hydroxyvalerate (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 9-heptadecyl alcohol (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0388] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0389] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0390] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 30.

[0391] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.56-4.11(m,8H),2.43-2.33(m, 12H),2.34(s,6H),1.94-1.81(m,4H),1.68-1.20(m,60H),0.92-0.88(m,12H).

[0392] Example 9: Synthesis of Compound 31

[0393] 6-Hydroxyhexanoic acid (1.5 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 9-heptadecyl alcohol (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0394] Intermediate 1 (2.2 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.2 eq), 4-dimethylaminopyridine (1.0 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0395] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0396] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 31.

[0397] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.56-4.11(m,8H),2.43-2.33(m, 12H),2.34(s,6H),1.94-1.81(m,4H),1.68-1.20(m,64H),0.92-0.88(m,12H).

[0398] Example 10: Synthesis of Compound 39

[0399] 2-Butyloctanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an empty container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0400] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0401] 9-Heptadecyl alcohol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 6-hydroxyhexanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0402] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0403] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0404] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 39.

[0405] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.57-4.13(m,7H),2.43-2.33(m, 10H),2.34(s,6H),1.98-1.76(m,3H),1.68-1.20(m,56H),0.92-0.88(m,12H).

[0406] Example 11: Synthesis of Compound 40

[0407] 2-Butyloctanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,6-hexanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an open container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0408] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0409] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0410] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0411] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0412] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 40.

[0413] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.36-4.19(m,8H),2.53-2.25(m, 8H),2.24(s,6H),1.98-1.76(m,4H),1.68-1.20(m,54H),0.92-0.88(m,12H).

[0414] Example 12: Synthesis of Compound 45

[0415] 2-Butyloctanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,6-hexanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an open container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0416] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0417] 9-Heptadecyl alcohol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 6-hydroxyhexanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0418] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0419] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0420] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 45.

[0421] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.57-4.13(m,7H),2.43-2.33(m, 10H),2.34(s,6H),1.98-1.76(m,3H),1.68-1.20(m,58H),0.92-0.88(m,12H).

[0422] Example 13: Synthesis of Compound 50

[0423] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0424] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0425] 9-Heptadecyl alcohol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 6-hydroxyhexanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0426] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0427] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0428] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 50.

[0429] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.57-4.13(m,7H),2.43-2.33(m, 10H),2.34(s,6H),1.98-1.76(m,3H),1.68-1.20(m,64H),0.92-0.88(m,12H).

[0430] Example 14: Synthesis of Compound 56

[0431] 2-Butyloctanol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 6-hydroxyhexanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0432] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0433] 9-Heptadecyl alcohol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 6-hydroxyhexanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0434] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0435] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0436] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 56.

[0437] 1H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.30-4.13(m,8H),2.53-2.33(m, 12H),2.34(s,6H),1.98-1.76(m,4H),1.68-1.20(m,52H),0.92-0.88(m,12H).

[0438] Example 15: Synthesis of Compound 61

[0439] 2-Butyloctanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,6-hexanediol (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an open container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0440] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0441] 3-Undecyl alcohol (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 8-hydroxyoctanoic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0442] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, intermediate 3 (3.0 eq) was added and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 4.

[0443] Intermediate 4 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water, and the mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 5.

[0444] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 5 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 61.

[0445] 1 H-NMR (400MHz, CDCl3): δ5.58-5.53(m,1H),4.52-4.13(m,7H),2.53-2.33(m, 10H),2.34(s,6H),1.98-1.76(m,3H),1.68-1.20(m,50H),0.92-0.88(m,12H).

[0446] Example 16: Synthesis of Compound 77

[0447] 3-Oxoglutaric acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, linolenic acid (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 1.

[0448] Intermediate 1 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. An appropriate amount of ethyl acetate and water were added for extraction. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 2.

[0449] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 2 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 77.

[0450] 1 H-NMR (400MHz, CDCl3): δ5.92-5.90(m,1H),5.64-5.53(m,8H),4.20-4.13(m,4H),2.85 -2.33(m,20H),2.16(s,6H),1.88-1.76(m,2H),1.68-1.20(m,36H),0.92-0.88(m,6H).

[0451] Example 17: Synthesis of Compound 81

[0452] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediamine (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an empty container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0453] Intermediate 1 (3.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, 3-oxoglutaric acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0454] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in tetrahydrofuran. The mixture was stirred in an ice bath for 30 minutes, and sodium borohydride (2.0 eq) was slowly added. After the reaction was brought back to room temperature, the reaction was continued for 2 hours. TLC monitoring showed that a new product was formed. The reaction was quenched by slowly adding ice water. The mixture was extracted with an appropriate amount of ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 3.

[0455] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 3 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 81.

[0456] 1 H-NMR (400MHz, CDCl3): δ5.26-5.21(m,1H),3.05-2.84(m,10H),2.44-2.13(m ,8H),2.04(s,6H),1.98-1.86(m,2H),1.68-1.20(m,60H),0.92-0.88(m,12H).

[0457] Example 18: Synthesis of Compound 91

[0458] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediamine (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an empty container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0459] Intermediate 1 (3.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, N-Cbz-β-glutamic acid (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The product was extracted with ethyl acetate and water, dried, concentrated, and purified by column chromatography. The product Pd / C was hydrogenated, filtered, and concentrated to obtain intermediate 2.

[0460] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 2 (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain compound 91.

[0461] 1 H-NMR (400MHz, CDCl3): δ5.26-5.21(m,1H),3.05-2.84(m,10H),2.44-2.13(m ,8H),2.04(s,6H),1.98-1.86(m,2H),1.68-1.20(m,60H),0.92-0.88(m,12H).

[0462] Example 19: Synthesis of Compound 100

[0463] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, 1,5-pentanediamine (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an empty container, concentrated, and purified by column chromatography to obtain intermediate 1.

[0464] Intermediate 1 (2.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, trimalonic acid (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0465] N,N-dimethyl-1,3-diaminopropane (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 2 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 100.

[0466] 1 H-NMR (400MHz, CDCl3): δ3.68-3.26(m,11H),2.44-2.13(m,14H),1.98-1.20(m,62H),0.92-0.88(m,12H).

[0467] Example 20: Synthesis of Compound 109

[0468] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, N-Cbz-5-amino-1-pentanol (3.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The product was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography. The product Pd / C was hydrogenated, filtered, and concentrated to obtain intermediate 1.

[0469] Intermediate 1 (3.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, N-Cbz-β-glutamic acid (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The product was extracted with ethyl acetate and water, dried, concentrated, and purified by column chromatography. The product Pd / C was hydrogenated, filtered, and concentrated to obtain intermediate 2.

[0470] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 2 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 109.

[0471] 1 H-NMR (400MHz, CDCl3): δ4.26-4.17(m,5H),3.08-2.95(m,6H),2.44-2.13(m,12H),1.98-1.20(m,64H),0.92-0.88(m,12H).

[0472] Example 21: Synthesis of Compound 118

[0473] 5-Amino-1-pentanol (1.0 eq) was added to a 250 mL single-necked flask under a nitrogen atmosphere and dissolved in anhydrous dichloromethane. Then, tert-butyldimethylchlorosilane (1.2 eq) was added at 0 °C, and the mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried, concentrated, and purified by column chromatography to obtain intermediate 1.

[0474] 2-Hexyldecanoic acid (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (2.5 eq) were added. After stirring for 30 minutes, intermediate 1 (3.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain intermediate 2.

[0475] Intermediate 2 (1.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1 M hydrogen fluoride acetonitrile solution (1.2 eq) was added dropwise. The mixture was stirred while monitoring for 15 minutes until the reaction was complete. The mixture was extracted with ethyl acetate and water, dried over the organic phase, concentrated, and purified by column chromatography to obtain intermediate 3.

[0476] Intermediate 3 (3.0 eq) was added to a 250 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (1.0 eq) were added. After stirring for 30 minutes, N-Cbz-β-glutamic acid (1.0 eq) was added. The reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and purified by column chromatography to obtain intermediate 4.

[0477] 4-Dimethylaminobutyrate (1.0 eq) was added to a 100 mL single-necked flask and dissolved in dichloromethane. Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.5 eq), 4-dimethylaminopyridine (0.5 eq), and triethylamine (3.0 eq) were added. After stirring for 30 minutes, intermediate 4 (1.0 eq) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate and water, dried over an organic phase, concentrated, and purified by column chromatography to obtain compound 118.

[0478] 1H-NMR (400MHz, CDCl3): δ4.56-4.47(m,1H),4.26-4.17(m,4H),3.28-3.04(m,6H),2.44-2.13(m,12H),1.98-1.20(m,64H),0.92-0.88(m,12H).

[0479] Example 22

[0480] The compounds in Table 1, along with cholesterol, DSPC (distearate phosphatidylcholine), and PEG-DMG (polyethylene glycol dimyristate), were dissolved in ethanol at a molar ratio of 50:38.5:10:1.5 (where the equivalent of the compounds was 50, the equivalent of cholesterol was 38.5, the equivalent of DSPC was 10, and the equivalent of PEG-DMG was 1.5) (concentration of 24.4 mg / mL based on total lipid weight). Luciferase mRNA was dissolved in 10 mM citrate buffered saline solution at pH 4.0 (drug concentration of 0.276 mg / mL) at a volume ratio of 1:3 (where the equivalent of the ethanol solution was 1 and the equivalent of the aqueous solution was 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with PBS at pH 7.4 using dialysis or tangential flow to remove the ethanol, thus preparing multiple groups of LNP@luci-mRNA.

[0481] The particle size, PDI, and encapsulation efficiency of each LNP@luci-mRNA were tested, and the results are shown in Table 1.

[0482] Table 1. Particle size, PDI, Zeta, and encapsulation efficiency of each LNP@luci-mRNA

[0483] compound Particle size (nm) PDI Zeta(mV) Encapsulation efficiency (%) 4 182 0.01 0.9 75 9 98 0.13 2.2 94 21 108 0.10 0.3 90 45 87 0.06 0.7 92 81 126 0.09 1.8 92 91 113 0.12 2.1 96 100 103 0.08 2.6 90 109 138 0.14 1.4 91

[0484] As shown in Table 1, the lipid nanoparticles formed by the above compounds have controllable particle size, uniform size distribution, electrically neutral surface, and high encapsulation efficiency.

[0485] Example 23

[0486] The three groups of LNP@luci-mRNAs formed from compounds 4, 21, and 45 in Example 22 were injected intramuscularly into Balb / c mice. Fluorescence imaging was performed 6 hours later. The injection dose of each group of LNP@luci-mRNA was 10 micrograms per mouse. The imaging results are as follows: Figure 1-5As shown, the results indicated that the LNP@luci-mRNA formed by compound 4 exhibited significant fluorescent expression at the injection site in mice, but its fluorescent expression in the liver and spleen was weak. In contrast, the LNP@luci-mRNA formed by compounds 21 and 45 showed significant fluorescent expression at the injection site, as well as in the liver and spleen of mice.

[0487] Example 24

[0488] Compound 12 was dissolved in ethanol (24.4 mg / mL, total lipid weight) in a molar ratio of 30:20:38.5:10:1.5 with DOTAP ((2,3-dioleopropyl)trimethylammonium chloride), cholesterol, DSPC, and PEG-DMG (where the equivalent of compound 12 was 30, the equivalent of DOTAP was 20, the equivalent of cholesterol was 38.5, the equivalent of DSPC was 10, and the equivalent of PEG-DMG was 1.5). Luciferase mRNA was dissolved in 50 mM citrate buffered saline solution at pH 4.0 (drug concentration 0.276 mg / mL) at a volume ratio of 1:3 (where the equivalent of the ethanol solution was 1 and the equivalent of the aqueous solution was 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with PBS at pH 7.4 using dialysis or tangential flow technology to prepare LNP@luci-mRNA. Adding sucrose as a cryoprotectant yields a nucleic acid lipid nanoparticle drug formulation.

[0489] Example 25

[0490] Compound 19 was dissolved in ethanol (24.4 mg / mL, total lipid weight) at a molar ratio of 20:25:15:25:5:10 (where the equivalent of compound 19 was 20, the equivalent of DOTAP was 25, the equivalent of DOPS was 15, the equivalent of cholesterol was 25, the equivalent of DSPC was 5, and the equivalent of PEG-DMG was 10). Luciferase mRNA (5 mg) was dissolved in a 50 mM citrate buffer solution at pH 4.0 (drug concentration 0.276 mg / mL). The volume ratio of the two solutions was 1:3 (where the equivalent of the ethanol solution was 1, and the equivalent of the aqueous solution was 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with a pH of 15 using dialysis or tangential flow techniques. LNPluci-@mRNA was prepared using PBS at 7.4°C. Sucrose was added as a cryoprotectant to obtain a nucleic acid lipid nanoparticle drug formulation.

[0491] Example 26

[0492] Compound 27 was dissolved in ethanol (concentration 24.4 mg / mL, based on total lipid weight) with DLin-KC2-DMA (CAS No.: 1190197-97-7), DOPG (dioleoylphosphatidylglycerol), cholesterol, DSPC, and Tween-80 (total 30 mg) in a molar ratio of 15:5:3:51.5:25:0.5 (where the equivalent of compound 27 is 15, the equivalent of DLin-KC2-DMA is 5, the equivalent of DOPG is 3, the equivalent of cholesterol is 51.5, the equivalent of DSPC is 25, and the equivalent of Tween-80 is 0.5). Luciferase mRNA (1 mg) was dissolved in pH [a solution / concentration]. LNP@luci-mRNA was prepared by rapidly mixing the two phases (drug concentration 0.276 mg / mL) in a 50 mM citrate buffer solution at a volume ratio of 1:3 (where the ethanol equivalent was 1 and the aqueous equivalent was 3). The buffer environment was then replaced with PBS at pH 7.4 using dialysis or tangential flow techniques. Sucrose was added as a cryoprotectant to obtain the nucleic acid lipid nanoparticle drug formulation.

[0493] It should be noted that although specific examples have been used to illustrate the technical solutions of the present invention, those skilled in the art will understand that the present invention is not limited thereto. Various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, and are not limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical applications, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A compound of formula (I-2) or a pharmaceutically acceptable salt thereof, in: R1 and R2 are each independently 2-butyloct-1-yl, 2-butyldec-1-yl, 2-hexylhex-1-yl, 2-hexyloct-1-yl, 2-hexyldec-1-yl, tridecane-7-yl, pentadecane-7-yl, heptadecan-9-yl, or nonadecan-9-yl; B1 and B2 are each independently C5 straight-chain alkylene groups; B3 is a C3 straight-chain alkylene group; X is .

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, R1 and R2 are each independently 2-butyloct-1-yl, 2-hexyldec-1-yl, pentadecane-7-yl, heptadecan-9-yl, or nonadecan-9-yl.

3. The following compounds or their pharmaceutically acceptable salts: 。 4. A lipid carrier comprising the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof; The lipid carrier comprises a first lipid compound and a second lipid compound, wherein, The first lipid compound comprises the compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof and optionally a cationic lipid, and the second lipid compound comprises one or more of anionic lipids, neutral lipids, sterols and amphiphilic lipids.

5. The lipid carrier according to claim 4, characterized in that, The cationic lipids include one or more of DLinDMA, DODMA, DLin-MC2-MPZ, DLin-KC2-DMA, DOTAP, C12-200, DC-Chol and DOTMA. The anionic lipids include one or more of phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, DOPG, DOPS and myristoyl phosphatidylglycerol; The neutral lipids include at least one of DOPE, DSPC, DPPC, DOPC, DPPG, POPC, POPE, DPPE, DMPE, DSPE and SOPE, or lipids modified with anionic or cationic modifying groups. The amphiphilic lipids include one or more of PEG-DMG, PEG-c-DMG, PEG-C14, PEG-c-DMA, PEG-DSPE, PEG-PE, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, Tween-20, Tween-80, PEG-DPG, PEG-s-DMG, DAA, PEG-c-DOMG, and GalNAc-PEG-DSG.

6. The lipid carrier according to claim 5, characterized in that, The cationic lipids include DLin-KC2-DMA and DOTAP.

7. The lipid carrier according to claim 5, characterized in that, The anionic lipids include DOPG and DOPS.

8. The lipid carrier according to claim 5, characterized in that, The neutral lipids include DSPC.

9. The lipid carrier according to claim 5, characterized in that, The amphiphilic lipids include PEG-DMG and Tween-80.

10. The lipid carrier according to claim 4, characterized in that, The molar ratio of the first lipid compound, the anionic lipid, the neutral lipid, the sterol, and the amphiphilic lipid is (20~65):(0~20):(5~25):(25~55):(0.3~15); wherein, in the first lipid compound, the molar ratio of the compound according to any one of claims 1-3 or its pharmaceutically acceptable salt to the cationic lipid is (1~10):(0~10).

11. The lipid carrier according to claim 10, characterized in that, The molar ratio of the first lipid compound, the anionic lipid, the neutral lipid, the sterol, and the amphiphilic lipid is (20~55):(0~13):(5~25):(25~51.5):(0.5~15); wherein, in the first lipid compound, the molar ratio of the compound or its pharmaceutically acceptable salt according to any one of claims 1-3 to the cationic lipid is (3~4):(0~5).

12. A nucleic acid nanoparticle composition comprising a compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof, or a lipid carrier according to any one of claims 4-11, and a nucleic acid drug.

13. The nucleic acid nanoparticle composition according to claim 12, characterized in that, The nucleic acid drug includes one or more of the following: DNA, siRNA, mRNA, dsRNA, antisense nucleic acid, antisense oligonucleotide, microRNA, antisense microRNA, antagomir, microRNA inhibitor, microRNA activator, and immunostimulatory nucleic acid.

14. The nucleic acid nanoparticle composition according to claim 12 or 13, characterized in that, The mass ratio of the nucleic acid drug to the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3 is 1:(3-40); or The mass ratio of the nucleic acid drug to the lipid carrier according to any one of claims 4-11 is 1:(3~40).

15. A pharmaceutical formulation comprising a compound according to any one of claims 1-3 or a pharmaceutically acceptable salt thereof, or a lipid carrier according to any one of claims 4-11, or a nucleic acid lipid nanoparticle composition according to any one of claims 12-14, and a pharmaceutically acceptable excipient, carrier, and diluent.

16. The pharmaceutical preparation according to claim 15, characterized in that, The particle size of the pharmaceutical preparation is 30~500 nm.

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