Composition and compound for delivering therapeutic agent

Abstract

A lipid nanoparticle composition for delivering a therapeutic agent, comprising a compound of formula I or compounds of formula I and formula II, or a salt thereof.

Description

Compositions and compounds for delivery of therapeutic agents Technical Field

[0001] The present disclosure belongs to the field of medicine and relates to a composition and compound for delivering a therapeutic agent. Background Art

[0002] Nucleic acid-based drugs, such as messenger RNA (mRNA), antisense oligonucleotides, small interfering RNA (siRNA), plasmids, etc., have broad application prospects. How to safely and effectively deliver them to target organs and target cells in the body is a difficult problem that restricts the development of this technology.

[0003] Currently, nucleic acid drug delivery systems can be divided into viral vector systems and non-viral systems. Lipid nanoparticle-mediated nucleic acid drug delivery is the main method of non-viral delivery systems.

[0004] Lipid nanoparticles (LNPs) have been demonstrated as excellent carriers of nucleic acids for treating various diseases in gene therapy and vaccine applications. Composed of cationic lipids and other auxiliary lipids, such as cholesterol, phospholipids, and PEGylated lipids, LNPs encapsulate nucleic acids, protecting them from degradation, promoting cellular uptake, and reducing immune responses. Furthermore, LNPs offer other advantages for cellular delivery of bioactive ingredients, including good targeting, minimal side effects, excellent stability, and high transfection efficiency.

[0005] Historically, effective LNPs are composed of 4 components: ionizable cationic lipids, zwitterionic phospholipids, cholesterol and lipid poly (ethylene glycol) (PEG). However, these LNPs only result in the general delivery of nucleic acids, rather than the delivery of targeted organs or tissues. LNPs are usually only delivered to the liver by RNA. CN112930198A discloses a new class of LNPs, which add DOTAP (permanent cation) to prepare LNPs on the basis of 5A2-SC8 (ionizable cation), DOPE (zwitterionic), cholesterol, and DMG-PEG to achieve spleen and lung targeting. This application proves that DOTAP has specificity, and replacing DOTAP with other lipids cannot achieve corresponding effects.

[0006] The rapid development of the therapeutic field based on nucleic acid molecules has put forward greater demands on the delivery of nucleic acid drugs, and therefore it is necessary to develop efficient and safe nucleic acid delivery vectors.

[0007] Currently, the lipid compositions for delivering nucleic acid drugs that are clinically available can basically only target liver tissue or be administered locally. Therefore, the development of new nucleic acid drug delivery vectors that can achieve targeted delivery to other organs is of great significance to expanding the application of nucleic acid drugs. Summary of the Invention

[0008] The present disclosure provides a lipid nanoparticle composition comprising a compound represented by formula I or a salt thereof,

[0009] in,

[0010] L a and L b Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-;

[0011] H a 、H b and H c Each independently selected from C 1-10 alkylene;

[0012] R a and R b Each independently selected from C 1-24 Alkyl and C 2-24 alkenyl;

[0013] R c Selected from NR c1 R c2 and N + R c3 R c4 R c5 ;

[0014] R c1 and R c2 are each independently selected from hydrogen and C 1-6 alkyl;

[0015] R c3 、R c4 and R c5 Each independently selected from C 1-6 alkyl.

[0016] In some embodiments, only the following substituents or groups are defined, and the definitions of the substituents or groups not mentioned are the same as those described in any other embodiment (hereinafter referred to as "in some embodiments"), L a and L b Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-.

[0017] In some embodiments, L a and L b At least one (eg, one) selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-.

[0018] In some embodiments, L a and L b At least one (eg, one) selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, where C 1-6 The alkylene group is not n-propylene.

[0019] In some embodiments, L a Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L b Selected from -OC(O)- and -C(O)O-.

[0020] In some embodiments, L b Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L a Selected from -OC(O)- and -C(O)O-.

[0021] In some embodiments, L a and L b Each is independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-.

[0022] In some embodiments, L a and L b are each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-, and L a and L b At least one (e.g., one) is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, or -C(O)O(CH2)2O-.

[0023] In some embodiments, L a is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- or -C(O)O(CH2)2O-, L b Selected from -OC(O)- and -C(O)O-.

[0024] In some embodiments, L b is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- or -C(O)O(CH2)2O-, L a Selected from -OC(O)- and -C(O)O-.

[0025] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from linear or branched C 1-6 Alkylene.

[0026] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6C in alkylene-O- 1-6 The alkylene group is independently selected from C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene and C6 alkylene.

[0027] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from C 1-4 Alkylene.

[0028] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene groups are independently selected from methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene, isobutylene and tert-butylene.

[0029] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene groups are independently selected from methylene and ethylene.

[0030] In some embodiments, L a and L b In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene group is not n-propylene.

[0031] In some embodiments, H a Selected from linear or branched C1-10 In some embodiments, H a Selected from linear or branched C 3-8 In some embodiments, H a Selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-.

[0032] In some embodiments, H b Selected from linear or branched C 1-10 In some embodiments, H b Selected from linear or branched C 3-8 In some embodiments, H b Selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-.

[0033] In some embodiments, H c Selected from linear or branched C 1-10 In some embodiments, H c Selected from linear or branched C 1-6 In some embodiments, H c Selected from linear or branched C 1-4 In some embodiments, H c Selected from -CH2-, -(CH2)2-, -(CH2)3- and -(CH2)4-.

[0034] In some embodiments, H a 、H b and H c In the C 1-10 Each alkylene group is independently a straight chain C 1-10 Alkylene.

[0035] In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently branched C 1-10 Alkylene.

[0036] In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently selected from C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene and C 10Alkylene.

[0037] In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently selected from C 3-8 Alkylene.

[0038] In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently selected from C 1-6 In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently selected from C 1-4 In some embodiments, H a 、H b and H c In the C 1-10 The alkylene groups are each independently selected from methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene, isobutylene and tert-butylene.

[0039] In some embodiments, R a and R b Each independently selected from a linear or branched C 1-24 alkyl.

[0040] In some embodiments, R a and R b Each independently selected from a linear or branched C 4-18 alkyl.

[0041] In some embodiments, R a Selected from straight chain C 4-18 Alkyl, R b Selected from branched C 4-18 alkyl.

[0042] In some embodiments, R a Selected from branched C 4-18 Alkyl, R b Selected from branched C 4-18 alkyl.

[0043] In some embodiments, R a Selected from branched C 4-18 Alkyl, R b Selected from straight chain C 4-18 alkyl.

[0044] In some embodiments, R a and R b In the C 1-24 The alkyl groups are each independently selected from C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 Alkyl, C 19 Alkyl, C 20 Alkyl, C 21 Alkyl, C 22 Alkyl, C 23 Alkyl and C 24 In some embodiments, R a and R b In the C 1-24 The alkyl groups are each independently selected from C 4-18 alkyl.

[0045] In some embodiments, R a and R b In the C 1-24 The alkyl groups are each independently selected from linear C 1-24 Alkyl and branched C 3-24 alkyl.

[0046] In some embodiments, the linear C 1-24 Alkyl is selected from linear C 4-18 In some embodiments, the linear C 4-18 Alkyl is selected from C 7-13 alkyl.

[0047] In some embodiments, the branched chain C 3-24 Alkyl group is selected from branched C 4-18 In some embodiments, the branched C 4-18 Alkyl group is selected from branched C 9-18 In some embodiments, the branched C 3-24 Alkyl group is selected from Wherein, m1 and m2 are each independently an integer from 0 to 12, and the sum of m1 and m2 does not exceed 21.

[0048] In some embodiments, R a and R b Each independently selected from:

[0049] In some embodiments, R a and R b Each independently selected from branched C 4-18 Alkenyl.

[0050] In some embodiments, R a and R b Each independently selected from a linear C 4-18 Alkenyl.

[0051] In some embodiments, R a Selected from straight chain C 4-18 Alkenyl, R b Selected from branched C 4-18 Alkenyl.

[0052] In some embodiments, R a Selected from branched C 4-18 Alkenyl, R b Selected from branched C 4-18 Alkenyl.

[0053] In some embodiments, R a and R b In the C 2-24 Each alkenyl group independently includes, but is not limited to, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C 10 Alkenyl, C 11 Alkenyl, C 12 Alkenyl, C 13 Alkenyl, C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 17 Alkenyl, C 18 Alkenyl, C 19 Alkenyl, C 20 Alkenyl, C 21 Alkenyl, C 22 Alkenyl, C 23 Alkenyl, C 24 In some embodiments, R a and R b In the C 2-24 Alkenyl is selected from C 4-18 Alkenyl.

[0054] In some embodiments, R a and R b In the C 2-24 Each alkenyl group is independently selected from a straight chain C 2-24 Alkenyl and branched C 4-24 Alkenyl.

[0055] In some embodiments, the linear C 2-24 Alkenyl is selected from linear C 4-18 Alkenyl.

[0056] In some embodiments, the branched chain C 4-24 Alkenyl is selected from branched C 4-18 Alkenyl.

[0057] In some embodiments, R c Selected from NR c1 R c2 .

[0058] In some embodiments, R c Selected from NH2.

[0059] In some embodiments, R c Selected from NHR c1 and NHR c2 , R c1 and R c2 Each independently selected from C 1-6 In some embodiments, R c Selected from NHCH3.

[0060] In some embodiments, R c1 and R c2 In the C 1-6 The alkyl groups are each independently selected from C 1-4 In some embodiments, R c1 and R c2 In the C 1-6 Each alkyl group is independently selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. c1 and R c2 In the C 1-6 The alkyl group is selected from methyl.

[0061] In some embodiments, R c Selected from N + R c3 R c4 R c5 In some embodiments, R c Selected from N + (CH3)3.

[0062] In some embodiments, R c Selected from NH2, NHCH3 and N + (CH3)3.

[0063] In some embodiments, Rc3 、R c4 and R c5 In the C 1-6 The alkyl groups are each independently selected from C 1-4 In some embodiments, R c3 、R c4 and R c5 In the C 1-6 Each alkyl group is independently selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. c3 、R c4 and R c5 In the C 1-6 The alkyl group is selected from methyl.

[0064] In some embodiments, the compound represented by Formula I or a salt thereof is a compound represented by Formula Ia or a salt thereof,

[0065] Among them, L a , L b 、H a 、H b 、H c 、R a and R b As defined in any one of the embodiments of the compound shown in Formula I.

[0066] In some embodiments, the compound of formula I or its salt is selected from the compounds of formula IA, IB, IC, ID, IE and IF or their salts.

[0067] Among them, H c 、R a 、R b and R c As defined in any one of the embodiments of the compound shown in Formula I;

[0068] R d1 、R d2 、R d3 、R d4 、R d5 、R d6 、R e1 、R e2 、R e3 、R e4 、R e5 、R e6 、R f1 、R f2 、R f3 、R f4 、R f5 、R f6 、R g1、R g2 、R g3 、R g4 、R g5 、R g6 、R h3 、R h4 、R h5 、R h6 、R i3 、R i4 、R i5 and R i6 are each independently selected from hydrogen and C 1-6 alkyl;

[0069] n1, n2, n3, n4, n5, n6, n8, n9, n11, n12, n14 and n15 are each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

[0070] n7, n10, n13 and n16 are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

[0071] In some embodiments, the compound represented by formula I or a salt thereof is selected from the compound represented by formula IC or a salt thereof.

[0072] In some embodiments, R d1 、R d2 、R d3 、R d4 、R d5 、R d6 、R e1 、R e2 、R e3 、R e4 、R e5 、R e6 、R f1 、R f2 、R f3 、R f4 、R f5 、R f6 、R g1 、R g2 、R g3 、R g4 、R g5 、R g6 、R h3 、R h4 、R h5 、R h6 、R i3 、R i4 、R i5 and R i6 Selected from hydrogen.

[0073] In some embodiments, n1, n2, n3, n4, n5, n6, n8, n9, n11, n12, n14 and n15 are each independently selected from 3, 4, 5, 6 and 7.

[0074] In some embodiments, n7, n10, n13, and n16 are each independently selected from 1, 2, 4, 5, and 6.

[0075] In some embodiments, n7, n10, n13, and n16 are each independently selected from 1 and 2.

[0076] In some embodiments, the compound represented by Formula I is selected from the following structures,

[0077] In some embodiments, the lipid nanoparticle composition further comprises a compound of Formula II or a salt thereof,

[0078] Among them, L 1 and L 2 Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, and L 1 and L 2 At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-;

[0079] H 1 、H 2 and H 3 Each independently selected from C 1-10 alkylene;

[0080] R 1 and R 2 Each independently selected from C 1-24 Alkyl and C 2-24 alkenyl;

[0081] R 3 Selected from hydroxyl groups.

[0082] In some embodiments, L 1 and L2 Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-.

[0083] In some embodiments, L 1 Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L 2 Selected from -OC(O)- and -C(O)O-.

[0084] In some embodiments, L 2 Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L 1 Selected from -OC(O)- and -C(O)O-.

[0085] In some embodiments, L 1 and L 2 Each is independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH(CH3)C(O)O-, -OC(O)CH(CH3)O-, -OC(CH3)2C(O)O-, -OC(O)C(CH3)2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, -C(O)O(CH2)2O-, -O(CH2)4OC(O)- and -C(O)O(CH2)4O-.

[0086] In some embodiments, L 1 and L 2are each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH(CH3)C(O)O-, -OC(O)CH(CH3)O-, -OC(CH3)2C(O)O-, -OC(O)C(CH3)2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, -C(O)O(CH2)2O-, -O(CH2)4OC(O)- and -C(O)O(CH2)4O-, and L 1 and L 2 At least one of them is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH(CH3)C(O)O-, -OC(O)CH(CH3)O-, -OC(CH3)2C(O)O-, -OC(O)C(CH3)2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, -C(O)O(CH2)2O-, -O(CH2)4OC(O)- and -C(O)O(CH2)4O-.

[0087] In some embodiments, L 1 and L 2 In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from linear or branched C 1-6 Alkylene.

[0088] In some embodiments, L 1 and L 2 In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene group is independently selected from C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene and C6 alkylene.

[0089] In some embodiments, L 1 and L 2 In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from C 1-4 Alkylene.

[0090] In some embodiments, L 1 and L 2 In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene groups are independently selected from methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene, isobutylene and tert-butylene.

[0091] In some embodiments, L 1 and L 2 In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene groups are independently selected from methylene and ethylene.

[0092] In some embodiments, H 1 Selected from linear or branched C 1-10 In some embodiments, H 1 Selected from linear or branched C 3-8 In some embodiments, H 1 Selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-.

[0093] In some embodiments, H 2 Selected from linear or branched C 1-10 In some embodiments, H 2 Selected from linear or branched C 3-8 In some embodiments, H 2 Selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-.

[0094] In some embodiments, H 3 Selected from linear or branched C1-10 In some embodiments, H 3 Selected from linear or branched C 1-6 In some embodiments, H 3 Selected from linear or branched C 1-4 In some embodiments, H 3 Selected from -CH2-, -(CH2)2-, -(CH2)3- and -(CH2)4-.

[0095] In some embodiments, H 1 、H 2 and H 3 In the C 1-10 Each alkylene group is independently a straight chain C 1-10 Alkylene.

[0096] In some embodiments, H 1 、H 2 and H 3 In the C 1-10 The alkylene groups are each independently branched C 1-10 Alkylene.

[0097] In some embodiments, H 1 、H 2 and H 3 In the C 1-10 The alkylene groups are each independently selected from C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene and C 10 Alkylene.

[0098] In some embodiments, H 1 、H 2 and H 3 In the C 1-10 The alkylene groups are each independently selected from C 3-8 Alkylene.

[0099] In some embodiments, H 1 、H 2 and H 3 In the C 1-10 The alkylene groups are each independently selected from C 1-6 In some embodiments, H 1 、H 2 and H 3 In the C 1-10 The alkylene groups are each independently selected from C 1-4 In some embodiments, H 1 、H 2 and H 3In the C 1-10 The alkylene groups are each independently selected from methylene, ethylene, n-propylene, isopropylene, n-butylene, sec-butylene, isobutylene and tert-butylene.

[0100] In some embodiments, R 1 and R 2 Each independently selected from a linear or branched C 1-24 alkyl.

[0101] In some embodiments, R 1 and R 2 Each independently selected from a linear or branched C 4-18 alkyl.

[0102] In some embodiments, R 1 Selected from straight chain C 4-18 Alkyl, R 2 Selected from branched C 4-18 alkyl.

[0103] In some embodiments, R 1 Selected from branched C 4-18 Alkyl, R 2 Selected from branched C 4-18 alkyl.

[0104] In some embodiments, R 1 Selected from straight chain C 4-18 Alkyl, R 2 Selected from straight chain C 4-18 alkyl.

[0105] In some embodiments, R 1 and R 2 In the C 1-24 The alkyl groups independently include, but are not limited to, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 Alkyl, C 19 Alkyl, C 20 Alkyl, C 21 Alkyl, C 22 Alkyl, C 23 Alkyl, C 24 In some embodiments, R 1 and R 2In the C 1-24 The alkyl groups are each independently selected from C 4-18 alkyl.

[0106] In some embodiments, R 1 and R 2 In the C 1-24 The alkyl groups are each independently selected from linear C 1-24 Alkyl and branched C 3-24 alkyl.

[0107] In some embodiments, the linear C 1-24 Alkyl is selected from linear C 4-18 In some embodiments, the linear C 4-18 Alkyl is selected from C 7-13 alkyl.

[0108] In some embodiments, the branched chain C 3-24 Alkyl group is selected from branched C 4-18 In some embodiments, the branched C 4-18 Alkyl group is selected from branched C 9-18 In some embodiments, the branched C 3-24 Alkyl group is selected from Wherein, p1 and p2 are each independently an integer from 0 to 12, and the sum of p1 and p2 does not exceed 21.

[0109] In some embodiments, R 1 and R 2 Each independently selected from:

[0110] In some embodiments, R 1 and R 2 Each independently selected from branched C 4-18 Alkenyl.

[0111] In some embodiments, R 1 and R 2 Each independently selected from a linear C 4-18 Alkenyl.

[0112] In some embodiments, R 1 Selected from straight chain C 4-18 Alkenyl, R 2 Selected from branched C 4-18 Alkenyl.

[0113] In some embodiments, R 1 Selected from branched C 4-18 Alkenyl, R 2 Selected from branched C 4-18 Alkenyl.

[0114] In some embodiments, R 1 and R 2 In the C 2-24 Alkenyl includes but is not limited to C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C 10 Alkenyl, C 11 Alkenyl, C 12 Alkenyl, C 13 Alkenyl, C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 17 Alkenyl, C 18 Alkenyl, C 19 Alkenyl, C 20 Alkenyl, C 21 Alkenyl, C 22 Alkenyl, C 23 Alkenyl, C 24 In some embodiments, R 1 and R 2 In the C 2-24 Alkenyl is selected from C 4-18 Alkenyl.

[0115] In some embodiments, R 1 and R 2 In the C 2-24 Alkenyl is selected from linear C 2-24 Alkenyl and branched C 4-24 Alkenyl.

[0116] In some embodiments, the linear C 2-24 Alkenyl is selected from linear C 4-18 Alkenyl.

[0117] In some embodiments, the branched chain C 4-24 Alkenyl is selected from branched C 4-18 Alkenyl.

[0118] In some embodiments, the compound represented by formula II or its salt is selected from the compounds represented by formula IIA, IIB, IIC and IID or their salts.

[0119] Among them, H 3 、R 1 、R 2 and R 3 As defined in any one of the schemes of the compound shown in formula II;

[0120] R 4a 、R 4b 、R 4c 、R 4d、R 5a 、R 5b 、R 5c 、R 5d 、R 6a 、R 6b 、R 6c 、R 6d 、R 7a 、R 7b 、R 7c 、R 7d 、R 8a 、R 8b 、R 8c 、R 8d 、R 9a 、R 9b 、R 9c and R 9d are each independently selected from hydrogen and C 1-6 alkyl;

[0121] o1, o2, o4, o5, o7, o8, o10 and o11 are each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

[0122] o3, o6, o9 and o12 are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

[0123] In some embodiments, the compound represented by formula II or a salt thereof is selected from the compound represented by formula IIA or a salt thereof.

[0124] In some embodiments, R 4a 、R 4b 、R 4c 、R 4d 、R 5a 、R 5b 、R 5c 、R 5d 、R 6a 、R 6b 、R 6c 、R 6d 、R 7a 、R 7b 、R 7c 、R 7d 、R 8a 、R 8b 、R 8c 、R 8d 、R 9a 、R 9b 、R 9c and R 9d Selected from hydrogen.

[0125] In some embodiments, o1, o2, o4, o5, o7, o8, o10, and o11 are each independently selected from 2, 3, 4, and 5.

[0126] In some embodiments, o3, o6, o9, and o12 are each independently selected from 1, 2, 3, and 4.

[0127] In some embodiments, o3 is selected from 1.

[0128] In some embodiments, o3 is selected from 1, R 4a and R 5a are each independently selected from hydrogen and methyl.

[0129] In some embodiments, the compound represented by formula II is selected from compound II-1,

[0130] In some embodiments, the molar percentage of the compound of Formula I or its salt is 5% to 80% (e.g., 5%, 10%, 11.25%, 12%, 15%, 17.5%, 20%, 24%, 25%, 30%, 35%, 36%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80% and any value therebetween). In some embodiments, the molar percentage of the compound of Formula I or its salt is 10% to 80% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 10% to 60% (e.g., 11.25%, 12%, 17.5%, 24%, 25%, 36% or 50%) of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 10% to 40% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 10% to 30% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 15% to 60% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 15% to 35% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of Formula I or its salt is 40% to 60% of the lipid nanoparticle composition.

[0131] In some embodiments, the molar percentage of the compound of formula II or its salt is 5% to 50% (e.g., 5%, 10%, 12%, 15%, 20%, 24%, 25%, 30%, 35%, 36%, 37.5%, 40%, 43.75%, 45%, 50%, and any value therebetween). In some embodiments, the molar percentage of the compound of formula II or its salt is 15% to 35% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of formula II or its salt is 10% to 45% (e.g., 12%, 24%, 25%, 30%, 36%, 37.5%, or 43.75%) of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of formula II or its salt is 20% to 45% of the lipid nanoparticle composition.

[0132] In some embodiments, the lipid nanoparticle composition comprises a phospholipid.

[0133] In some embodiments, the phospholipid is selected from 1,2-dipalmitoyl-sn-glycero-3-o-4'-(N,N,N-trimethyl)homoserine (DGTS), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, the phospholipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).

[0134] In some embodiments, the phospholipid comprises 1% to 25% by mole of the lipid nanoparticle composition (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, and any number therebetween). In some embodiments, the phospholipid comprises 1% to 15% by mole of the lipid nanoparticle composition (e.g., 5% or 10%). In some embodiments, the phospholipid comprises 1% to 8% by mole of the lipid nanoparticle composition. In some embodiments, the phospholipid comprises 8% to 15% by mole of the lipid nanoparticle composition.

[0135] In some embodiments, the lipid nanoparticle composition further comprises a steroid.

[0136] In some embodiments, the steroid is selected from cholesterol.

[0137] In some embodiments, the steroid comprises 10% to 50% by mole of the lipid nanoparticle composition (e.g., 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 29.5%, 30%. %, 48.5%, 49%, 49.5%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, In some embodiments, the steroid comprises 10% to 45% (e.g., 18.5%, 38.5%, or 40.5%) of the lipid nanoparticle composition by mole. In some embodiments, the steroid comprises 10% to 25% of the lipid nanoparticle composition by mole. In some embodiments, the steroid comprises 30% to 45% of the lipid nanoparticle composition by mole.

[0138] In some embodiments, the lipid nanoparticle composition further comprises a PEGylated lipid.

[0139] In some embodiments, the PEGylated lipid comprises a PEG component of about 1000 to about 10,000 Daltons (e.g., 1000 Daltons, 2000 Daltons, 3000 Daltons, 4000 Daltons, 5000 Daltons, 6000 Daltons, 7000 Daltons, 8000 Daltons, 9000 Daltons, or 10000 Daltons).

[0140] In some embodiments, the PEGylated lipid is selected from PEGylated diacylglycerols. In some embodiments, the PEGylated lipid is selected from 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG-PEG). In some embodiments, the PEGylated lipid is DMG-PEG 2000.

[0141] In some embodiments, the PEGylated lipid comprises 0.5% to 10% (e.g., 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%) of the lipid nanoparticle composition by mole. In some embodiments, the PEGylated lipid comprises 0.5% to 5% (e.g., 1.5%) of the lipid nanoparticle composition by mole.

[0142] In some embodiments, the lipid nanoparticle composition comprises the compound of formula I or a salt thereof, a phospholipid, a steroid, and a pegylated lipid.

[0143] In some embodiments, the lipid nanoparticle composition comprises the compound of formula I or a salt thereof, a phospholipid, a steroid and a pegylated lipid; the compound of formula I or its salt accounts for 10% to 80% by mole of the lipid nanoparticle composition; the phospholipid is selected from DGTS, DOPE and DSPC, and the phospholipid accounts for 1% to 25% by mole of the lipid nanoparticle composition; the steroid is selected from cholesterol, and the steroid accounts for 10% to 50% by mole of the lipid nanoparticle composition; the pegylated lipid is selected from pegylated diacylglycerol, and the pegylated lipid accounts for 0.5% to 10% by mole of the lipid nanoparticle composition.

[0144] In some embodiments, the lipid nanoparticle composition comprises the compound of formula Ia or a salt thereof, a phospholipid, a steroid and a pegylated lipid; the compound of formula Ia or its salt accounts for 10% to 80% by mole of the lipid nanoparticle composition; the phospholipid is selected from DGTS, DOPE and DSPC, and the phospholipid accounts for 1% to 25% by mole of the lipid nanoparticle composition; the steroid is selected from cholesterol, and the steroid accounts for 10% to 50% by mole of the lipid nanoparticle composition; the pegylated lipid is selected from pegylated diacylglycerol, and the pegylated lipid accounts for 0.5% to 10% by mole of the lipid nanoparticle composition.

[0145] In some embodiments, the lipid nanoparticle composition comprises the compound represented by formula Ia or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar percentage of the compound represented by formula Ia or its salt is 15% to 60% of the lipid nanoparticle composition; the molar percentage of the DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of the cholesterol is 30% to 45% of the lipid nanoparticle composition; and the molar percentage of the DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0146] In some embodiments, the lipid nanoparticle composition comprises any one of the above-mentioned compounds I-1 to I-10 or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar percentage of any one of the compounds I-1 to I-10 or a salt thereof is 15% to 60% of the lipid nanoparticle composition; the molar percentage of the DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of the cholesterol is 30% to 45% of the lipid nanoparticle composition; and the molar percentage of the DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0147] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-1 or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar ratio of the compound I-1 or a salt thereof, DSPC, cholesterol and DMG-PEG is 50:10:38.5:1.5.

[0148] In some embodiments, the lipid nanoparticle composition comprises the compound of formula I or its salt, the compound of formula II or its salt, phospholipids, steroids and pegylated lipids.

[0149] In some embodiments, the lipid nanoparticle composition comprises the compound of formula I or its salt, the compound of formula II or its salt, a phospholipid, a steroid and a pegylated lipid; the molar percentage of the compound of formula I or its salt is 10% to 80% of the lipid nanoparticle composition; the molar percentage of the compound of formula II or its salt is 5% to 50% of the lipid nanoparticle composition; the phospholipid is selected from DGTS, DOPE and DSPC, and the molar percentage of the phospholipid is 1% to 25% of the lipid nanoparticle composition; the steroid is selected from cholesterol, and the molar percentage of the steroid is 10% to 50% of the lipid nanoparticle composition; the pegylated lipid is selected from pegylated diacylglycerol, and the molar percentage of the pegylated lipid is 0.5% to 10% of the lipid nanoparticle composition.

[0150] In some embodiments, the lipid nanoparticle composition comprises the compound of formula I or its salt, the compound of formula II or its salt, DSPC, cholesterol and DMG-PEG; the molar percentage of the compound of formula I or its salt is 15% to 60% of the lipid nanoparticle composition; the molar percentage of the compound of formula II or its salt is 15% to 35% of the lipid nanoparticle composition; the molar percentage of DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of cholesterol is 10% to 25% or 30% to 45% of the lipid nanoparticle composition; and the molar percentage of DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0151] In some embodiments, the lipid nanoparticle composition comprises the compound represented by formula Ia or a salt thereof, the compound represented by formula II or a salt thereof (e.g., compound II-1 or a salt thereof), DSPC, cholesterol and DMG-PEG; the molar percentage of the compound represented by formula I or its salt is 10% to 60% of the lipid nanoparticle composition; the molar percentage of the compound represented by formula II or its salt is 10% to 45% of the lipid nanoparticle composition; the molar percentage of DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of cholesterol is 10% to 45% of the lipid nanoparticle composition; and the molar percentage of DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0152] In some embodiments, the lipid nanoparticle composition comprises the compound represented by formula Ia or a salt thereof, the compound represented by formula II or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar percentage of the compound represented by formula I or its salt is 15% to 60% of the lipid nanoparticle composition; the molar percentage of the compound represented by formula II or its salt is 15% to 35% of the lipid nanoparticle composition; the molar percentage of DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of cholesterol is 10% to 25% or 30% to 45% of the lipid nanoparticle composition; and the molar percentage of DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0153] In some embodiments, the lipid nanoparticle composition comprises the compound represented by formula Ia or a salt thereof, the compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG; the compound represented by formula Ia or its salt accounts for 15% to 60% by mole of the lipid nanoparticle composition; the compound II-1 or its salt accounts for 15% to 35% by mole of the lipid nanoparticle composition; the DSPC accounts for 1% to 15% by mole of the lipid nanoparticle composition; the cholesterol accounts for 10% to 25% or 30% to 45% by mole of the lipid nanoparticle composition; and the DMG-PEG accounts for 0.5% to 5% by mole of the lipid nanoparticle composition.

[0154] In some embodiments, the lipid nanoparticle composition comprises any one of the above-mentioned compounds I-1 to I-10 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar percentage of any one of the compounds I-1 to I-10 or a salt thereof is 10% to 60% of the lipid nanoparticle composition; the molar percentage of the compound II-1 or a salt thereof is 10% to 45% of the lipid nanoparticle composition; the molar percentage of the DSPC is 1% to 15% of the lipid nanoparticle composition; the molar percentage of the cholesterol is 10% to 45% of the lipid nanoparticle composition; and the molar percentage of the DMG-PEG is 0.5% to 5% of the lipid nanoparticle composition.

[0155] In some embodiments, the lipid nanoparticle composition comprises any one of the above-mentioned compounds I-1 to I-10 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG; the molar percentage of any one of the compounds I-1 to I-10 or a salt thereof in the lipid nanoparticle composition is 15% to 60%; the molar percentage of the compound II-1 or its salt in the lipid nanoparticle composition is 15% to 35%; the molar percentage of the DSPC in the lipid nanoparticle composition is 1% to 15%; the molar percentage of the cholesterol in the lipid nanoparticle composition is 10% to 25% or 30% to 45%; the molar percentage of the DMG-PEG in the lipid nanoparticle composition is 0.5% to 5%.

[0156] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-1 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG, and the molar ratio of the compound I-1 or a salt thereof, compound II-1, DSPC, cholesterol and DMG-PEG is 25:25:10:38.5:1.5 or 50:25:5:18.5:1.5.

[0157] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-1 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG, and the molar ratio of the compound I-1 or a salt thereof, compound II-1, DSPC, cholesterol and DMG-PEG is 11.25:43.75:5:38.5:1.5, 12:36:10:40.5:1.5, 17.5:37.5:5:38.5:1.5, 24:24:10:40.5:1.5, 25:30:5:38.5:1.5 or 36:12:10:40.5:1.5.

[0158] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-2 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG, and the molar ratio of the compound I-2 or a salt thereof, compound II-1, DSPC, cholesterol and DMG-PEG is 25:25:10:38.5:1.5 or 50:25:5:18.5:1.5.

[0159] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-8 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG, and the molar ratio of the compound I-8 or a salt thereof, compound II-1, DSPC, cholesterol and DMG-PEG is 50:25:5:18.5:1.5.

[0160] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned compound I-10 or a salt thereof, the above-mentioned compound II-1 or a salt thereof, DSPC, cholesterol and DMG-PEG, and the molar ratio of the compound I-10 or a salt thereof, compound II-1, DSPC, cholesterol and DMG-PEG is 50:25:5:18.5:1.5.

[0161] The present disclosure provides a lipid nanoparticle composition comprising a primary amine lipid.

[0162] In some embodiments, the primary amine lipid is a compound represented by formula Ia or a salt thereof as described in any of the above schemes,

[0163] Among them, L a 、L b 、H a 、H b 、H c 、R a and R b As defined in any of the above embodiments of the compound represented by formula I.

[0164] In some embodiments, the compound represented by Formula Ia is selected from any one of the following compounds,

[0165] In some embodiments, the lipid nanoparticle composition further comprises a compound of formula II or a salt thereof as described in any of the above schemes,

[0166] Among them, L 1 、L 2 、H 1 、H 2 、H 3 、R 1 、R 2 and R 3 As defined in any of the preceding schemes.

[0167] In some embodiments, the compound represented by formula II is selected from compound II-1,

[0168] In some embodiments, the primary amine lipid comprises 5% to 80% by mole of the lipid nanoparticle composition (e.g., 5%, 10%, 11.25%, 12%, 15%, 17.5%, 20%, 24%, 25%, 30%, 35%, 36%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, and any number therebetween). In some embodiments, the primary amine lipid comprises 10% to 80% by mole of the lipid nanoparticle composition. In some embodiments, the primary amine lipid comprises 10% to 60% by mole of the lipid nanoparticle composition (e.g., 11.25%, 12%, 17.5%, 24%, 25%, 36%, or 50%). In some embodiments, the primary amine lipid comprises 10% to 40% by mole of the lipid nanoparticle composition. In some embodiments, the primary amine lipid comprises 10% to 30% by mole of the lipid nanoparticle composition. In some embodiments, the primary amine lipid comprises 15% to 60% by mole of the lipid nanoparticle composition. In some embodiments, the primary amine lipid comprises 15% to 35% by mole of the lipid nanoparticle composition. In some embodiments, the primary amine lipid comprises 40% to 60% by mole of the lipid nanoparticle composition.

[0169] In some embodiments, the molar percentage of the compound of formula II or its salt is 5% to 50% (e.g., 5%, 10%, 12%, 15%, 20%, 24%, 25%, 30%, 35%, 36%, 37.5%, 40%, 43.75%, 45%, 50%, and any value therebetween). In some embodiments, the molar percentage of the compound of formula II or its salt is 15% to 35% of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of formula II or its salt is 10% to 45% (e.g., 12%, 24%, 25%, 30%, 36%, 37.5%, or 43.75%) of the lipid nanoparticle composition. In some embodiments, the molar percentage of the compound of formula II or its salt is 20% to 45% of the lipid nanoparticle composition.

[0170] In some embodiments, the lipid nanoparticle composition comprises a phospholipid.

[0171] In some embodiments, the phospholipid is selected from 1,2-dipalmitoyl-sn-glycero-3-o-4'-(N,N,N-trimethyl)homoserine (DGTS), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, the phospholipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).

[0172] In some embodiments, the phospholipid comprises 1% to 25% by mole of the lipid nanoparticle composition (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, and any number therebetween). In some embodiments, the phospholipid comprises 1% to 15% by mole of the lipid nanoparticle composition (e.g., 5% or 10%). In some embodiments, the phospholipid comprises 1% to 8% by mole of the lipid nanoparticle composition. In some embodiments, the phospholipid comprises 8% to 15% by mole of the lipid nanoparticle composition.

[0173] In some embodiments, the lipid nanoparticle composition further comprises a steroid.

[0174] In some embodiments, the steroid is selected from cholesterol.

[0175] In some embodiments, the steroid comprises 10% to 50% by mole of the lipid nanoparticle composition (e.g., 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 29.5%, 30%. %, 48.5%, 49%, 49.5%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, In some embodiments, the steroid comprises 10% to 45% (e.g., 18.5%, 38.5%, or 40.5%) of the lipid nanoparticle composition by mole. In some embodiments, the steroid comprises 10% to 25% of the lipid nanoparticle composition by mole. In some embodiments, the steroid comprises 30% to 45% of the lipid nanoparticle composition by mole.

[0176] In some embodiments, the lipid nanoparticle composition further comprises a PEGylated lipid.

[0177] In some embodiments, the PEGylated lipid comprises a PEG component of about 1000 to about 10,000 Daltons (e.g., 1000 Daltons, 2000 Daltons, 3000 Daltons, 4000 Daltons, 5000 Daltons, 6000 Daltons, 7000 Daltons, 8000 Daltons, 9000 Daltons, or 10000 Daltons).

[0178] In some embodiments, the PEGylated lipid is selected from PEGylated diacylglycerols. In some embodiments, the PEGylated lipid is selected from 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG-PEG). In some embodiments, the PEGylated lipid is DMG-PEG 2000.

[0179] In some embodiments, the PEGylated lipid comprises 0.5% to 10% (e.g., 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%) of the lipid nanoparticle composition by mole. In some embodiments, the PEGylated lipid comprises 0.5% to 5% (e.g., 1.5%) of the lipid nanoparticle composition by mole.

[0180] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned primary amine lipids, phospholipids, steroids and pegylated lipids; the primary amine lipids account for 10% to 80% of the lipid nanoparticle composition by mole; the phospholipids are selected from DGTS, DOPE and DSPC, and the phospholipids account for 1% to 25% of the lipid nanoparticle composition by mole; the steroid is selected from cholesterol, and the steroid accounts for 10% to 50% of the lipid nanoparticle composition by mole; the pegylated lipid is selected from pegylated diacylglycerol, and the pegylated lipid accounts for 0.5% to 10% of the lipid nanoparticle composition by mole.

[0181] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned primary amine lipid, DSPC, cholesterol and DMG-PEG; the primary amine lipid accounts for 15% to 60% of the lipid nanoparticle composition by mole; the DSPC accounts for 1% to 15% of the lipid nanoparticle composition by mole; the cholesterol accounts for 30% to 45% of the lipid nanoparticle composition by mole; and the DMG-PEG accounts for 0.5% to 5% of the lipid nanoparticle composition by mole.

[0182] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned primary amine lipid, the above-mentioned compound shown in Formula II, a phospholipid, a steroid and a pegylated lipid; the primary amine lipid accounts for 10% to 80% of the lipid nanoparticle composition by mole; the compound shown in Formula II or its salt accounts for 5% to 50% of the lipid nanoparticle composition by mole; the phospholipid is selected from DGTS, DOPE and DSPC, and the phospholipid accounts for 1% to 25% of the lipid nanoparticle composition by mole; the steroid is selected from cholesterol, and the steroid accounts for 10% to 50% of the lipid nanoparticle composition by mole; the pegylated lipid is selected from pegylated diacylglycerol, and the pegylated lipid accounts for 0.5% to 10% of the lipid nanoparticle composition by mole.

[0183] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned primary amine lipid, the above-mentioned compound shown in Formula II, DSPC, cholesterol and DMG-PEG; the primary amine lipid accounts for 15% to 60% of the lipid nanoparticle composition by mole; the compound shown in Formula II or its salt accounts for 15% to 35% of the lipid nanoparticle composition by mole; the DSPC accounts for 1% to 15% of the lipid nanoparticle composition by mole; the cholesterol accounts for 10% to 25% or 30% to 45% of the lipid nanoparticle composition by mole; the DMG-PEG accounts for 0.5% to 5% of the lipid nanoparticle composition by mole.

[0184] In some embodiments, the lipid nanoparticle composition comprises the above-mentioned primary amine lipid, the compound shown in formula II, DSPC, cholesterol and DMG-PEG; the primary amine lipid accounts for 10% to 60% of the lipid nanoparticle composition by mole; the compound shown in formula II or its salt accounts for 10% to 45% of the lipid nanoparticle composition by mole; the DSPC accounts for 1% to 15% of the lipid nanoparticle composition by mole; the cholesterol accounts for 10% to 45% of the lipid nanoparticle composition by mole; the DMG-PEG accounts for 0.5% to 5% of the lipid nanoparticle composition by mole.

[0185] In some embodiments, the lipid nanoparticle composition of the present disclosure further comprises a therapeutic agent selected from the group consisting of a small molecule, a protein, and a nucleic acid.

[0186] In some embodiments, the therapeutic agent is selected from nucleic acids.

[0187] In some embodiments, the nucleic acid is selected from a therapeutic nucleic acid.

[0188] In some embodiments, the nucleic acid is selected from siRNA, miRNA, pri-miRNA, messenger RNA (mRNA), clustered regularly interspaced short palindromic repeats (CRISPR)-related nucleic acids, single guide RNA (sgRNA), CRISPR-RNA (crRNA), trans-activating crRNA (tracrRNA), plasmid DNA (pDNA), transfer RNA (tRNA), antisense oligonucleotide (ASO), guide RNA (gRNA), double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), single-stranded RNA (ssRNA) and double-stranded RNA (dsRNA). In some embodiments, the nucleic acid is selected from mRNA.

[0189] In some embodiments, the weight ratio of the lipid nanoparticle composition to the nucleic acid is about 1:1 to about 100:1.

[0190] In some embodiments, the compositions described herein preferentially deliver the therapeutic agent to a target organ selected from the group consisting of the lung, heart, liver, spleen, or stomach.

[0191] In some embodiments, the target organ is the lung.

[0192] In some embodiments, the therapeutic agent is distributed more highly in the lungs than in other organs.

[0193] The present disclosure provides a compound represented by formula III or a salt thereof,

[0194] in,

[0195] L A and L B Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, and L A and L B At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, where C 1-6 The alkylene group is not n-propylene;

[0196] H a 、H b 、H c 、R a 、R b and R c As defined in any one of the embodiments of the compound of formula I.

[0197] In some embodiments, L A and L B Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC1-6 Alkylene-O-, where C 1-6 The alkylene group is not n-propylene.

[0198] In some embodiments, L A and L B At least one (eg, one) selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-.

[0199] In some embodiments, L A Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L B Selected from -OC(O)- and -C(O)O-, wherein C 1-6 The alkylene group is not n-propylene.

[0200] In some embodiments, L B Selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, L A Selected from -OC(O)- and -C(O)O-, wherein C 1-6 The alkylene group is not n-propylene.

[0201] In some embodiments, L A and L B Each is independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-.

[0202] In some embodiments, L A and L Bare each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-, and L A and L B At least one (e.g., one) is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, or -C(O)O(CH2)2O-.

[0203] In some embodiments, L A and L B In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from linear or branched C 1-6 Alkylene.

[0204] In some embodiments, L A and L B In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene group is independently selected from C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene and C6 alkylene.

[0205] In some embodiments, L A and L B In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene is independently selected from C 1-4 Alkylene.

[0206] In some embodiments, L A and L B In, -OC 1-6Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 The alkylene groups are independently selected from methylene, ethylene, isopropylene, n-butylene, sec-butylene, isobutylene and tert-butylene.

[0207] In some embodiments, L A and L B In, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 C in alkylene-O- 1-6 Alkylene groups are independently selected from methylene and ethylene.

[0208] In some embodiments, the compound represented by formula III or its salt is selected from the compound represented by formula IIIa or its salt.

[0209] Among them, L A 、L B 、H a 、H b 、H c 、R a and R b As defined in any of the embodiments of the compound of formula III.

[0210] In some embodiments, the compound represented by formula III or a salt thereof is selected from the compounds represented by formula IIIA, IIIB, IIIC and IIID or a salt thereof,

[0211] Among them, H c 、R a 、R b and R c As defined in any one of the embodiments of the compound of formula I;

[0212] R d3 、R d4 、R d5 、R d6 、R e3 、R e4 、R e5 、R e6 、R f3 、R f4 、R f5 、R f6 、Rg3 、R g4 、R g5 、R g6 、R h3 、R h4 、R h5 、R h6 、R i3 、R i4 、R i5 、R i6 As defined in any one of the compounds of formula IC, ID, IE and IF;

[0213] n5, n6, n7, n8, n9, n10, n11, n12, n13, n14, n15 and n16 are as defined in any of the embodiments of the compounds of Formula IC, ID, IE and IF.

[0214] In some embodiments, the compound represented by Formula III or its salt is a compound or its salt having the following structure:

[0215] The present disclosure provides an isotope substitution of the above-mentioned compound or its salt. In some embodiments, the isotope substitution is a deuterium substitution.

[0216] The present disclosure provides a pharmaceutical composition comprising the lipid nanoparticle composition or the compound or its salt or the isotope substitution described above, and a pharmaceutically acceptable excipient.

[0217] In some embodiments, the aforementioned lipid nanoparticle composition, the aforementioned compound or salt thereof, or the aforementioned isotope substitution is in a therapeutically effective amount.

[0218] In some embodiments, the pharmaceutical composition contains 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the pharmaceutical composition. In some embodiments, the pharmaceutical composition contains 0.1% to 99.9% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 0.5% to 99.5% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 1% to 99% of a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition contains 2% to 98% of a pharmaceutically acceptable excipient.

[0219] The present disclosure also provides a use of the above-mentioned lipid nanoparticle composition, the above-mentioned compound or its salt, the above-mentioned isotope substitution, or the above-mentioned pharmaceutical composition in the preparation of a drug for inducing an immune response in a subject.

[0220] The present disclosure also provides a use of the above-mentioned lipid nanoparticle composition, the above-mentioned compound or its salt, the above-mentioned isotope substitution, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing and / or treating diseases or conditions related to polypeptide overexpression.

[0221] The present disclosure also provides a use of the above-mentioned lipid nanoparticle composition, the above-mentioned compound or its salt, the above-mentioned isotope substitution, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing and / or treating diseases or conditions related to insufficient polypeptide expression.

[0222] In some embodiments, the disease or condition includes, but is not limited to, cancer, infection, autoimmune disease, neurodegenerative disease, and inflammation.

[0223] The present disclosure also provides a use of the above-mentioned lipid nanoparticle composition, the above-mentioned compound or its salt, the above-mentioned isotope substitution, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing and / or treating cancer, infection, autoimmune disease, neurodegenerative disease and inflammation.

[0224] On the other hand, the present disclosure also provides a method for preventing and / or treating a disease or condition associated with inducing an immune response in a subject, comprising administering the above-mentioned lipid nanoparticle composition, or the above-mentioned compound or its salt, or the above-mentioned isotope substitute, or the above-mentioned pharmaceutical composition to the subject.

[0225] The present disclosure also provides a method for preventing and / or treating a disease or condition associated with polypeptide overexpression, comprising administering the lipid nanoparticle composition, the compound or its salt, the isotope substitution, or the pharmaceutical composition to a subject.

[0226] The present disclosure also provides a method for preventing and / or treating a disease or condition associated with insufficient polypeptide expression, comprising administering the lipid nanoparticle composition, the compound or its salt, the isotope substitution, or the pharmaceutical composition to a subject.

[0227] The present disclosure also provides a method for preventing and / or treating cancer, infection, autoimmune disease, neurodegenerative disease and inflammation, comprising administering the lipid nanoparticle composition, the compound or its salt, the isotope substitution or the pharmaceutical composition to a subject.

[0228] On the other hand, the present disclosure also provides the above-mentioned lipid nanoparticle composition or the above-mentioned compound or salt thereof or the above-mentioned isotope substitution or the above-mentioned pharmaceutical composition for preventing and / or treating a disease or condition associated with inducing an immune response in a subject.

[0229] The present disclosure also provides the lipid nanoparticle composition, the compound or its salt, the isotope substitution, or the pharmaceutical composition for preventing and / or treating diseases or disorders associated with polypeptide overexpression.

[0230] The present disclosure also provides the lipid nanoparticle composition, the compound or salt thereof, the isotope substitution, or the pharmaceutical composition for preventing and / or treating diseases or conditions associated with insufficient polypeptide expression.

[0231] The present disclosure also provides the lipid nanoparticle composition, the compound or its salt, the isotope substitution, or the pharmaceutical composition for preventing and / or treating cancer, infection, autoimmune disease, neurodegenerative disease, and inflammation.

[0232] The present disclosure also provides a use of the compound represented by formula I or a salt thereof or the compound represented by formula III or a salt thereof for preparing a lipid nanoparticle composition targeted to the lung.

[0233] The lipid nanoparticle compositions and compounds disclosed herein have good targeting properties to the lungs.

[0234] Definition of terms

[0235] The salts of the compounds described in the present disclosure include "acid" addition salts and "base" addition salts. For example, salts formed by acid-base reaction with a basic group (amino group), the acid including organic acid or inorganic acid.

[0236] The compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All of these isomers and their mixtures are included within the scope of the present disclosure. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

[0237] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary groups are cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and then the pure enantiomers are recovered. In addition, separation of enantiomers and diastereoisomers is typically accomplished using chromatography using a chiral stationary phase, optionally combined with chemical derivatization (e.g., to form carbamates from amines).

[0238] In the chemical structures of the compounds disclosed herein, the bond Indicates that the configuration is not specified, that is, if chiral isomers exist in the chemical structure, the bond Can be or include both Two configurations. In the chemical structure of the compound disclosed in the present invention, the bond The configuration is not specified, that is, it can be Z configuration or E configuration, or contain both configurations.

[0239] Unless otherwise specified, the symbols used in this paper are It means that it can be linked to one or more any groups according to the disclosure described herein.

[0240] The compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also referred to as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerizations. An example of a lactam-lactim equilibrium is between A and B as shown below:

[0241] All tautomeric forms are within the scope of the present disclosure. The naming of compounds does not exclude any tautomers.

[0242] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0243] Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have at least 1000 times the abundance of deuterium greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 10% deuterium incorporation). In the example, the compound has a natural abundance greater than deuterium that can be at least 1000 times the abundance of deuterium, at least 2000 times the abundance of deuterium, at least 3000 times the abundance of deuterium, at least 4000 times the abundance of deuterium, at least 5000 times the abundance of deuterium, at least 6000 times the abundance of deuterium or more abundant deuterium. The disclosure also includes various deuterated forms of formula (I) compounds. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of formula (I) compounds with reference to relevant literature. Commercially available deuterated starting materials may be used in the preparation of deuterated forms of the compounds of formula (I), or they may be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0244] "Optionally" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "C 1-6 The term "alkyl" means that halogen or cyano may but need not be present, and the description includes both the case where the alkyl is substituted by halogen or cyano and the case where the alkyl is not substituted by halogen and cyano.

[0245] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0246] "Pharmaceutically acceptable excipients" or "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the U.S. Food and Drug Administration for use in humans or domestic animals.

[0247] "Prevention" includes: (1) inhibiting the onset of a disease in a subject or patient who may be at risk for and / or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease; and / or (2) slowing the onset of symptoms or signs of a disease in a subject or patient who may be at risk for and / or susceptible to the disease but who does not yet experience or display any or all of the symptoms or signs of the disease.

[0248] "Treating" includes (1) inhibiting the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., arresting further development of the symptoms and / or signs), (2) ameliorating the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., reversing the symptoms and / or signs), and / or (3) achieving any measurable reduction in the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease.

[0249] As used herein, an "effective amount," "effective dose," "effective therapeutic amount," or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition being treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

[0250] As used herein, "subject," "patient," "subject," or "individual" are used interchangeably and include humans or non-human animals, such as mammals, eg, humans or monkeys.

[0251] In this disclosure, the terms "comprising", "including" or "containing" may be replaced with "consisting of".

[0252] The term "nucleic acid" is a polymer composed of nucleotides, such as deoxyribonucleotides (DNA) or ribonucleotides (RNA). Exemplary nucleic acids include, but are not limited to, deoxyribonucleotides (DNA), ribonucleic acids (RNA), including messenger RNA, RNAi-inducing agents, shRNA, siRNA, miRNA, antisense RNA, and the like.

[0253] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.

[0254] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group (C 1-24 In some embodiments, an alkyl group is a straight or branched chain group (C 3-24 In some embodiments, an alkyl group is a straight or branched chain group (C 4-18 In some embodiments, an alkyl group is a straight or branched chain group (C 9-18 In some embodiments, an alkyl group is a straight or branched chain group (C 7-13 In some embodiments, an alkyl group is a straight or branched chain group of 1 to 10 carbon atoms (C 1-10 In some embodiments, an alkyl group is a straight or branched chain group of 4 to 8 carbon atoms (C 4-8 In some embodiments, an alkyl group is a straight or branched chain group of 1 to 6 carbon atoms (C 1-6 In some embodiments, an alkyl group is a straight or branched chain group of 1 to 4 carbon atoms (C 1-4Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof.

[0255] The alkyl group may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0256] The term "alkylene" is the remaining group after removing one hydrogen atom from an "alkyl" group, wherein the definition of alkyl is as described above.

[0257] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing at least one carbon-carbon double bond, including straight-chain and branched alkenyl groups. In some embodiments, alkenyl is a straight-chain or branched alkenyl group (C 2-24 In some embodiments, alkenyl is a straight or branched alkenyl group (C 4-24 In some embodiments, alkenyl is a straight or branched alkenyl group of 4 to 18 carbon atoms (C 4-18 In some embodiments, alkenyl is a straight or branched alkenyl group of 9 to 18 carbon atoms (C 9-18 In some embodiments, alkenyl is a straight or branched alkenyl group of 7 to 13 carbon atoms (C 7-13 In some embodiments, alkenyl is a straight or branched alkenyl group of 2 to 10 carbon atoms (C2-10 In some embodiments, alkenyl is a straight or branched alkenyl group of 4 to 10 carbon atoms (C 4-10 In some embodiments, alkenyl is a straight or branched alkenyl group of 4 to 8 carbon atoms (C 4-8 Exemplary alkenyl groups include ethenyl, propenyl, n-butenyl, isobutenyl, 3-methylbut-2-enyl, n-pentenyl, heptenyl, octenyl, cyclohexyl-butenyl, and decenyl.

[0258] Alkenyl groups may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 7 membered cycloalkyl group.

[0259] The term "alkoxy" refers to an -O-alkyl group, wherein alkyl is as defined above.

[0260] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 8 carbon atoms, preferably 3 to 7 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, etc.; polycyclic cycloalkyls include spirocyclic, paracyclic, and bridged cycloalkyls. Cycloalkyls may be unsubstituted or substituted with one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 7 membered cycloalkyl group.

[0261] The terms "branched alkyl" and "branched alkenyl" refer to alkyl and alkenyl groups in which a non-terminal carbon atom of the carbon chain is bonded to at least one other carbon atom, and do not form a cyclic group, such as a spirocycle. Examples: C6 alkyl groups include: (straight chain), (branched chain) or (branched chain).

[0262] The term "hydroxy" refers to -OH.

[0263] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0264] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) whether substitution is possible or not without undue effort. BRIEF DESCRIPTION OF THE DRAWINGS

[0265] Figure 1 shows the fluorescence intensity of various tissues in mice injected with mRNA lipid nanoparticles through the tail vein. From left to right, each group includes heart, lung, liver, spleen, and kidney. * in Figure 1 indicates statistically significant differences, P < 0.05.

[0266] Figure 2 shows the percentage of protein expression in each tissue of mice injected with mRNA lipid nanoparticles through the tail vein. From left to right, each group includes heart, lung, liver, spleen, and kidney.

[0267] Figure 3 shows the fluorescence intensity of various tissues in mice after intratracheal injection of mRNA lipid nanoparticles. From left to right, each group is heart, lung, liver, spleen and kidney. In Figure 3, * represents statistically significant difference, P < 0.05.

[0268] Figure 4 shows the proportion of protein expression in each tissue of mice after intratracheal injection of mRNA lipid nanoparticles. From left to right, each group is heart, lung, liver, spleen and kidney.

[0269] FIG5 shows the detection of serum interleukin-6 (IL-6) concentration after mRNA lipid nanoparticles were injected into the tail vein of mice. ** in FIG5 represents a statistically significant difference, P<0.01.

[0270] FIG6 shows the detection of serum interleukin-6 (IL-6) concentration after intratracheal injection of mRNA lipid nanoparticles in mice. ** in FIG6 represents a statistically significant difference, P<0.01.

[0271] Figure 7 shows the encapsulation efficiency of LNP after atomization. In the figure, *** represents a statistically significant difference, P < 0.001.

[0272] Figure 8 shows the relative protein expression in the lungs of LNPs with different group ratios.

[0273] Figure 9 shows the fluorescence intensity of the lungs of mice injected with mRNA lipid nanoparticles through the tail vein. * in the figure represents a statistically significant difference, P < 0.05. DETAILED DESCRIPTION

[0274] The present disclosure is further described below with reference to examples, but these examples are not intended to limit the scope of the present disclosure.

[0275] Experimental methods in the examples disclosed herein that do not specify specific conditions are generally performed under conventional conditions or according to the conditions recommended by the raw material or product manufacturers. Reagents that do not specify their specific sources are conventional reagents purchased from the market.

[0276] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (Methanol-d4), with tetramethylsilane (TMS) as the internal standard.

[0277] HPLC determination used Agilent1100 high pressure liquid chromatograph, GAS15B DAD UV detector, Water Vbridge C18 150×4.6mm 5μm chromatographic column.

[0278] MS was determined using an Agilent 6120 triple quadrupole mass spectrometer, a G1315D DAD detector, and a Waters Xbridge C18 4.6×50 mm, 5 μm column. The mass was scanned in positive / negative ion mode with a mass scan range of 80 to 1200.

[0279] The thin layer chromatography silica gel plate used was Yantai Huanghai HSGF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) was 0.2 mm ± 0.03 mm. The specification used for thin layer chromatography separation and purification products was 0.4 mm - 0.5 mm.

[0280] The flash column purification system used was Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage).

[0281] Forward column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh or 300-400 mesh silica gel as the carrier, or uses Changzhou Santai pre-packed ultra-pure normal phase silica gel column (40-63μm, 60g, 24g, 40g, 120g or other specifications).

[0282] The known starting materials in the present disclosure can be synthesized by methods known in the art, or can be purchased from companies such as Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Bid Pharmaceuticals.

[0283] Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

[0284] Nitrogen atmosphere means that the reaction bottle is connected to a nitrogen balloon with a capacity of about 1L.

[0285] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0286] Hydrogen was produced by a QPH-1L hydrogen generator from Shanghai Quanpu Scientific Instrument Co., Ltd.

[0287] The nitrogen atmosphere or hydrogen atmosphere is usually evacuated and filled with nitrogen or hydrogen, and the operation is repeated three times.

[0288] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0289] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0290] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used to purify the compound, and the developing solvent system for thin layer chromatography, the volume ratio of the solvent were adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could be added for adjustment.

[0291] Example 1: Preparation of Compound I-1

[0292] Step 1:

[0293] Add water (46.8 mL) to a 500 mL three-necked flask. Cool to below 5°C and then add sodium hydroxide (46.8 g, 1170 mmol), maintaining the internal temperature below 15°C. Once the sodium hydroxide is completely dissolved, add toluene (46.8 mL) and compound 1a (9.71 g, 50.0 mmol, purchased from Adamas) and stir at room temperature. Cool again to below 5°C and add compound 1b (34.1 g, 175 mmol, purchased from Adamas) dropwise. Heat to room temperature in an oil bath and stir until the reaction is complete. The reaction solution was cooled to below 5°C, water (200 mL) was added, and the mixture was extracted with methyl tert-butyl ether (100 mL×3). The aqueous phase was cooled to below 5°C, concentrated hydrochloric acid was added until the pH value reached about 3, and then extracted with DCM (100 mL×5). The DCM phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (EA / PE=0-100%) to obtain 11.97 g of compound 1c (yield: 94%).

[0294] 1 H NMR (400MHz, CDCl3): δ8.50(brs,1H),7.36-7.26(m,5H),4.51(s,2H),4.13~ 4.10(m,2H),3.56(t,2H),3.48(t,2H),1.67~1.61(m,4H),1.50~1.43(m,2H).

[0295] Step 2:

[0296] Compound 1c (21.7 g, 86.0 mmol) was weighed into a reaction flask and added sequentially with DCM (434 mL), compound 1d (19.9 g, 77.4 mmol, prepared according to a reference method, Tetrahedron, 2011, vol. 67, #2, pp. 303-311), DMAP (10.5 g, 30.1 mmol), and EDCI (19.8 g, 103 mmol). The mixture was allowed to react overnight at room temperature. TLC monitored the reaction completion. Water (500 mL) was added to the reaction mixture, and the layers were separated. The aqueous phase was extracted with DCM (300 mL). The combined organic phases were washed with saturated sodium bicarbonate solution and then with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (EA / PE = 0-5%) to obtain 41.5 g of compound 1e (98% yield).

[0297] 1 H NMR (400MHz, CDCl3): δ7.34~7.33(m,4H),7.28~7.27(m,1H),4.97~4.95(m,1H),4.50(s,2H),4.04 (s,2H),3.54~3.46(m,4H),1.69~1.62(m,4H),1.58~1.43(m,2H),1.25(s,26H),0.89~0.83(m,8H).

[0298] Step 3:

[0299] Compound 1e (41.5 g, 84.3 mmol) was weighed and dissolved in THF (208 mL). Pd(OH)2 / C (4.15 g, 20%) was added and hydrogen was introduced via a double-layer hydrogen balloon. The mixture was stirred at room temperature for 5 hours. The reaction was monitored for completion by TLC. The mixture was filtered through Celite and the filter cake was washed with EA (100 mL x 3), concentrated under reduced pressure, and purified by column chromatography (EA / PE = 50%) to obtain 27.0 g of compound 1f (80% yield).

[0300] 1 H NMR (400MHz, CDCl3): δ4.98~4.93(m,1H),4.05(s,2H),3.66(t,2H),3.54(t, 2H), 1.70~1.58(m,4H), 1.53~1.43(m,4H), 1.25(s,26H), 0.89~0.86(m,6H).

[0301] Step 4:

[0302] Compound 1f (6.01 g, 15.0 mmol) was weighed and dissolved in extra-dry DCM (60 mL). Carbon tetrabromide (7.46 g, 22.5 mmol) was added under ice-bath. After stirring for 10 minutes, triphenylphosphine (5.51 g, 21.0 mmol) was added. After the addition, the mixture was stirred at room temperature until the reaction was complete. The mixture was purified by column chromatography (EA / PE = 0-5%) to obtain 6.75 g of compound 1g (yield 97%).

[0303] 1 H NMR (400MHz, CDCl3): δ4.99~4.93(m,1H),4.05(s,2H),3.54(t,2H),3.41(t,2H),1 .94~1.86(m,2H),1.70~1.63(m,2H),1.58~1.52(m,5H),1.26(s,25H),0.88(t,6H).

[0304] Step 5:

[0305] Compound 1g (1.0 g, 2.2 mmol) and compound 1h (1.9 g, 11.0 mmol, purchased from Anaiji) were dissolved in 20 mL of anhydrous ethanol and heated under a nitrogen atmosphere until the reaction was complete. The mixture was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography (DCM:MeOH = 10:1 to 5:1) to obtain 1.0 g of compound 1i (yield: 83%).

[0306] 1 H NMR (400MHz, CDCl3): δ5.13 (br s 1H),4.99~4.93(m,1H),4.04(s,2H),3.53(t,2H),3.25~3.16(m,2H),2.69(t,2H),2.63 (t,2H),1.71~1.61(m,4H),1.57~1.40(m,18H),1.33~1.19(m,24H),0.92~0.84(m,6H).

[0307] Step 6:

[0308] Compound 1i (0.487 g, 0.87 mmol), compound 1j (0.300 g, 0.795 mmol, prepared according to the method of patent WO2018 / 170306), potassium iodide (0.145 g, 0.87 mmol), and potassium carbonate (0.438 g, 3.018 mmol) were added to a 25 mL flask, and 5 mL each of cyclopentane methyl ether and anhydrous acetonitrile were added. Under a nitrogen atmosphere, the reaction was heated until the reaction was complete, cooled to room temperature, filtered, concentrated, and purified by column chromatography (PE:EA=10:1) to obtain 0.5 g of compound 1k (yield: 74%).

[0309] 1 H NMR (400MHz, CDCl3): δ5.60 (br s 1H),4.99~4.93(m,1H),4.84~4.78(m,1H),4.04(s,2H),3.53(t,2H),3.21 ~3.12(m,2H), 2.51~2.25(m,8H), 1.67~1.25(m,71H), 0.89~0.83(m,12H).

[0310] Step 7:

[0311] Compound 1k (0.5 g, 0.586 mmol) was dissolved in anhydrous dichloromethane (6.0 mL), and trifluoroacetic acid (2.0 mL) was added at room temperature and the reaction was almost complete. The reaction was concentrated under reduced pressure and purified by column chromatography (water:acetonitrile = 35%-95%) to obtain 370 mg of compound I-1 (yield: 84%).

[0312] 1 H NMR (400MHz, CDCl3): δ4.99~4.93(m,1H),4.84~4.78(m,1H),4.04(s,2H),3.51(t,2H) ,2.72(t,2H),2.46~2.35(m,6H),2.29(t,2H),1.67~1.25(m,62H),0.89~0.83(m,12H).

[0313] Example 2: Preparation of Compound I-2

[0314] Prepared with reference to the preparation method in WO2018170306A1.

[0315] Example 3: Preparation of Compound II-1

[0316] Prepared by referring to the preparation method in WO2023125738A.

[0317] Example 4: Preparation of Compound I-8

[0318] Step 1:

[0319] Compound 1g (1.0 g, 2.2 mmol) and 4b (2.0 g, 10.8 mmol, purchased from Anaiji) were dissolved in 50 mL of anhydrous ethanol and heated to an internal temperature of 60°C for 16 hours. The reaction mixture was then cooled to room temperature after heating. The reaction solution was concentrated and the crude product was purified by silica gel column chromatography (DCM:MeOH = 10:1-5:1) to afford approximately 0.3 g of 4c, in a yield of 27%.

[0320] 1 H NMR (400MHz, CDCl3): δ4.92~4.96(m,1H), 4.03(s,2H), 3.51~3.54(m,2H), 3.39~3.42(m,2H), 2 .84~2.93(m,7H), 2.17~2.29(m,2H), 1.88~1.92(m,2H), 1.25~1.70(m,42H), 0.86~0.89(m,6H).

[0321] Step 2:

[0322] 4c (0.23 g, 0.40 mmol), 1j (0.15 g, 0.40 mmol), potassium iodide (0.068 g, 0.40 mmol), and potassium carbonate (0.28 g, 2.01 mmol) were added to a 25 mL flask, and 5 mL each of cyclopentyl methyl ether and anhydrous acetonitrile were added. The mixture was heated to an external temperature of 86°C and reacted for 16 hours. The mixture was then cooled to room temperature, filtered, concentrated, and the crude product was subjected to column chromatography (PE:EA=10:1) to obtain approximately 0.1 g of 4d, in a yield of 28%.

[0323] 1 H NMR (400MHz, CDCl3): δ4.94~4.97(m,1H), 4.79~4.82(m,1H), 4.13(s,2H), 3.50~3.53(m,2H), 3.39~ 3.42(m,2H), 3.20(s,2H), 2.84(s,3H), 2.20~2.40(m,8H), 1.25~1.70(m,69H), 0.86~0.89(m,12H).

[0324] Step 3:

[0325] Compound 4d (0.1 g, 0.12 mmol) was dissolved in 4.0 mL of anhydrous dichloromethane. 1.0 mL of trifluoroacetic acid was added at room temperature. The reaction was allowed to react at room temperature for 4 hours. LC / MS control indicated the reaction was complete. Post-treatment: concentration was performed. The crude product was prepared by reverse phase ODS (water:acetonitrile = 35%-95%) to obtain approximately 80.3 mg of I-8 in a yield of 79%.

[0326] MS(ESI):m / z 767.6[M-CF3COOH+H] + .

[0327] 1H NMR (400MHz, CDCl3): δ4.90~4.94(m,1H), 4.78~4.82(m,1H), 4.03(s,2H), 3.52~3.55(m,2H), 3.39~3. 42(m,2H), 3.05~3.08(m,8H), 2.74(s,3H), 2.27~2.31(m,4H), 1.25~1.77(m,59H), 0.86~0.89(m,12H).

[0328] Example 5: Preparation of Compound I-10

[0329] Step 1:

[0330] Compound 5a (2.0 g, 9.2 mmol, prepared by the method of patent US2003 / 235916) was added to a flask, and anhydrous DCM (5 mL) was added and stirred to dissolve. 4 M HCl / dioxane solution (11.5 mL) was added at room temperature and reacted until the reaction was complete. The mixture was concentrated under reduced pressure to obtain 1.6 g of crude product, which was directly used in the next step.

[0331] 1 H NMR (400MHz, DMSO): δ8.24(br s,3H), 3.45~3.44(m,2H), 3.08(s,9H), 2.88~2.83(m,2H), 2.07~1.99(m,2H).

[0332] Step 2:

[0333] Compound 5b (197 mg, 0.70 mmol) and compound 4a (65 mg, 0.14 mmol) were added to a flask, and anhydrous ethanol (5 mL) was added and stirred to dissolve. Sodium carbonate (149 mg, 1.41 mmol) was added, and the reaction was heated under a nitrogen atmosphere until the reaction was complete. The mixture was filtered, concentrated, and purified by column chromatography (acetonitrile:methanol = 0-100%) to obtain 70 mg of compound 5c (yield: 80%).

[0334] 1 H NMR (400MHz, CDCl3): δ4.89~4.51(m,1H), 3.68~3.46(m,3H), 3.36~3.16(m,10H ), 2.31~2.21(m,2H), 1.98~1.78(m,2H), 1.63~1.17(m,41H), 0.92~0.82(m,6H).

[0335] Step 3:

[0336] Compound 5c (340 mg, 0.54 mmol) and compound 1g (252 mg, 0.54 mmol) were added to a flask, and potassium carbonate (375.5 g, 2.72 mmol) and potassium iodide (99.4 mg, 0.6 mmol) were added, and 5 mL each of anhydrous acetonitrile and cyclopentane methyl ether were added as solvents. Under a nitrogen atmosphere, the reaction was heated until the reaction was complete. The mixture was cooled to room temperature, filtered, concentrated, and purified by column chromatography (acetonitrile: methanol = 0-100%) to obtain 390 mg of compound I-10 (yield: 81%).

[0337] 1 H NMR (400MHz, MeOD): δ4.99~4.92(m,1H),4.88~4.83(m,1H),4.08(s,2H),3.56(t,2H),3.4~3.40(m,2H) ), 3.23~3.19(m,14H), 2.33~2.22(m,4H), 1.85~1.53(m,18H), 1.41~1.29(m,55H), 0.91~0.88(m,12H).

[0338] Example 6: Preparation of lipid nanoparticle composition

[0339] Preparation method

[0340] Compound I-1, compound I-2, compound II-1, compound I-8, compound I-10 and DOTAP (Ai Wei Tuo) are dissolved in ethanol solution (0.2mg / ml) respectively, respectively with DSPC, cholesterol, DMG-PEG (Ai Wei Tuo) solution dissolved in ethanol, with the molar percentage in table 1 mixed, prepare ethanol lipid solution.Luciferase mRNA (Luciferase, GenBank:MN728548.1) is dissolved in 50mM pH 5 acetic acid buffer, prepare mRNA aqueous solution (0.2mg / mL).Ethanol lipid solution and mRNA aqueous solution are mixed by microfluidics, ethanol phase and aqueous phase volume ratio is 1:3, and the weight ratio of total lipid and mRNA is about 20:1, prepares liposome.Ethanol is dialyzed out in 20mM Tris pH 7.5 solution, finally replaced in 20mM Tris pH 7.5 8% sucrose solution, obtain the lipid nanoparticle composition encapsulating mRNA.

[0341] Table 1

[0342] 1.2. Characterization of lipid nanoparticle composition

[0343] The size and polydispersity index (PDI) of lipid nanoparticles were determined by dynamic light scattering using a Malvern Zetasizer Pro in 173° backscattering detection mode.

[0344] The encapsulation efficiency was determined using the Quant-iT RiboGreen RNA Assay Kit.

[0345] The pKa values ​​of cations in liposome nanoparticles were determined using a fluorescence assay based on 6-(p-toluidine)-2-naphthalenesulfonic acid sodium salt (TNS). Buffer solutions of varying pH values ​​between 3 and 11.5 were prepared using 150 mM NaCl, 10 mM sodium phosphate, 10 mM sodium citrate, and 10 mM sodium borate. A 300 μM TNS solution was prepared and added to the buffer. Lipid nanoparticles were added to each buffer solution of varying pH values ​​and mixed thoroughly. Fluorescence intensity was measured at room temperature using a fluorescence microplate reader with an excitation wavelength of 325 nm and an emission wavelength of 435 nm. Fluorescence data were fitted and analyzed, and the pKa value was determined as the pH value that produces half-maximal fluorescence intensity.

[0346] Table 2

[0347] Test Example 1: Evaluation of mRNA delivery efficiency and tissue distribution of lipid nanoparticle compositions via tail vein injection in vivo

[0348] 6-8 week old female C57 BL / 6 mice (purchased from Shanghai Southern Model Organisms Science Co., Ltd.) were injected with a lipid nanoparticle composition encapsulating mRNA expressing luciferase (Luciferase, GenBank: MN728548.1) via the tail vein at a dose of 0.5 mg / kg. Six hours after administration, each mouse was intraperitoneally injected with luciferase substrate, and the heart, liver, spleen, lung, and kidney were dissected and taken using an IVIS small animal optical in vivo imaging instrument (PerkinElme). Fluorescence images were taken of the tissues and the fluorescence intensity of each tissue was counted. The level of fluorescence intensity represents the level of expression of luciferase protein, which reflects the efficiency of the lipid nanoparticle composition in delivering mRNA in vivo and its distribution in different organs and tissues. The average fluorescence intensity was calculated for at least three biological replicates of the lipid nanoparticles corresponding to each compound. The data are shown in Table 3 and Figure 1. The fluorescence intensity in Figure 1 is the fluorescence intensity corresponding to each tissue of the mouse captured and counted by the IVIS small animal optical in vivo imaging instrument. The fluorescence intensity represents the expression level of luciferase protein, which indicates the efficiency of mRNA delivery and tissue distribution of the lipid nanoparticle composition in vivo.

[0349] Using LNP-4 and LNP-8 as controls, the mRNA delivery efficiency and tissue distribution of lipid nanoparticle compositions corresponding to the composition LNP-1 containing compound I-1, the composition LNP-2 containing compound I-1 and compound II-1, and LNP-3 were detected after tail vein injection in mice.

[0350] Table 3

[0351] Table 4

[0352] Conclusion: As shown in Table 3 and Figure 1, the fluorescence intensity of the lipid nanoparticle composition LNP1-3 composed of Compound I-1 was the highest in the lungs, indicating the highest expression of luciferase protein in the lungs. Unlike traditional lipid nanoparticle compositions, such as the clinically available siRNA lipid nanoparticles that deliver siRNA to the liver, lipid nanoparticle compositions containing Compound I-1 deliver more nucleic acid drugs to the lungs. Furthermore, the protein expression in the lungs of the lipid nanoparticle composition LNP-3 composed of Compound I-1 was approximately 10 times that of the lipid nanoparticle composition LNP-4 composed of DOTAP.

[0353] As shown in Table 4 and Figure 2, the proportion of fluorescence intensity, which represents the proportion of protein expression in various organ tissues, shows that the proportion of lipid nanoparticle compositions LNP1-3 composed of compound I-1 in the lungs is higher than that in the liver and spleen, of which LNP-3 accounts for more than 90% in the lungs. The lipid nanoparticle composition LNP-3 containing compound I-1 has a higher proportion of protein expression in the lungs than the lipid nanoparticle composition LNP-4 composed of DOTAP. The lipid nanoparticle composition LNP-8 corresponding to compound II-1 is mainly distributed in the liver, with a distribution in the lungs of less than 1%.

[0354] Test Example 2: Evaluation of mRNA delivery efficiency and tissue distribution of lipid nanoparticle compositions via intratracheal administration

[0355] 6-8 week old female C57 BL / 6 mice (purchased from Shanghai South Model Organisms Science Co., Ltd.) were intratracheally administered with a lipid nanoparticle composition encapsulating mRNA expressing luciferase at a dose of 0.25 mg / kg. Six hours after administration, each mouse was intraperitoneally injected with luciferase substrate. The heart, liver, spleen, lung, and kidney were dissected and the tissues were imaged using an IVIS small animal optical in vivo imaging instrument (PerkinElme). Fluorescence images were taken of the tissues and the fluorescence intensity of each tissue was calculated. The fluorescence intensity represents the expression level of luciferase protein, reflecting the efficiency of the lipid nanoparticle composition in delivering mRNA in vivo and its distribution in different organs and tissues. The average fluorescence intensity was calculated for at least three biological replicates of the lipid nanoparticle composition corresponding to each compound, and the data are shown in Table 5. The percentage of the average fluorescence intensity of each group to the total fluorescence intensity was calculated to assess the tissue distribution of the protein expression of the lipid nanoparticle composition, as shown in Table 6. The fluorescence intensity in Figure 3 is the fluorescence intensity corresponding to each tissue of the mouse captured and counted by the IVIS small animal optical in vivo imaging instrument. The fluorescence intensity represents the expression level of luciferase protein, which indicates the efficiency of mRNA delivery and tissue distribution of the lipid nanoparticle composition in vivo.

[0356] Using a lipid nanoparticle composition containing DOTAP (LNP-4) as a control, the protein expression and tissue distribution of lipid nanoparticle compositions LNP-1 and LNP-2 containing compound I-1 were detected in the mouse respiratory tract under administration conditions.

[0357] Table 5

[0358] Table 6

[0359] Conclusion: As shown in Table 5 and Figure 3, the fluorescence intensity of lipid nanoparticle compositions composed of Compound I-1, LNP-1 and LNP-3, was the highest in the lungs, indicating the highest expression of luciferase protein in the lungs. Furthermore, the protein expression in the lungs of the lipid nanoparticle composition composed of Compound I-1, LNP-1, was approximately 32 times that of the lipid nanoparticle composition composed of DOTAP, LNP-4.

[0360] As shown in Table 6 and Figure 4, the fluorescence intensity ratio represents the ratio of protein expression in various organs and tissues. The lipid nanoparticle compositions LNP-1 and LNP-3 composed of compound I-1 have a higher ratio in the lungs than in the liver and spleen, of which LNP-1 has a ratio in the lungs exceeding 90%. The lipid nanoparticle compositions LNP-1 and LNP-3 containing compound I-1 have a higher ratio of protein expression in the lungs than the lipid nanoparticle composition LNP-4 composed of DOTAP. The fluorescence intensity and fluorescence intensity ratio of LNP-1 in the lungs are higher than those of LNP-8. The fluorescence intensity of LNP-8 in the liver is much higher than that of LNP-1 and LNP-3. This indicates that compound I-1 has greater lung expression among the LNP components and has better targeting.

[0361] Test Example 3: Evaluation of the in vivo safety of lipid nanoparticle compositions

[0362] Foreign substances entering the mammalian body trigger an innate immune response, thereby promoting the production of cytokines. These substances, upon entry, trigger an inflammatory response, which can easily lead to adverse reactions such as fever and edema. Therefore, the immunogenicity of lipid nanoparticle compositions in vivo was assessed by evaluating the concentration of cytokines, such as interleukin-6 (IL-6), in the blood of mice after injection with the lipid nanoparticle compositions. Lower cytokine concentrations indicate that the lipid nanoparticle compositions have lower immunogenicity and, therefore, better biosafety.

[0363] 6-8 week old BALB / c female mice (purchased from Shanghai Model Organisms Science Co., Ltd.) were injected via the tail vein at a dose of 0.5 mg / kg with lipid nanoparticle compositions encapsulating mRNA expressing a luciferase reporter gene. Six hours later, blood was collected for serum isolation. Serum IL-6 concentrations were measured using a mouse IL-6 ELISA kit. Serum IL-6 concentrations were measured in at least three biological replicates for each lipid nanoparticle composition, and the mean value was calculated.

[0364] Table 7

[0365] Conclusion: As shown in Table 7 and Figure 5, the IL-6 concentration in mouse serum of the LNP-3 lipid nanoparticle composition was significantly lower than that of the control lipid nanoparticle composition LNP-4. This indicates that the lipid nanoparticle composition containing compound I-1 has lower immunogenicity and better biosafety in vivo than the lipid nanoparticle composition corresponding to the control compound DOTAP.

[0366] 6-8 week old female BALB / c mice were intratracheally injected with lipid nanoparticle compositions containing mRNA expressing a luciferase reporter gene at a dose of 0.25 mg / kg. Blood was collected 6 hours later for serum isolation. Serum IL-6 concentrations were measured using a mouse IL-6 ELISA kit. Serum IL-6 concentrations were measured in at least three biological replicates for each lipid nanoparticle composition, and the mean value was calculated.

[0367] Table 8

[0368] Conclusion: As shown in Table 8 and Figure 6, the IL-6 concentration in mouse serum of the LNP-1 lipid nanoparticle composition was significantly lower than that of the control lipid nanoparticle composition LNP-4. This indicates that the lipid nanoparticle composition containing compound I-1 has lower immunogenicity and better biosafety in vivo than the lipid nanoparticle composition corresponding to the control compound DOTAP.

[0369] Test Example 4: Evaluation of Characterization Parameters of Lipid Nanoparticle Compositions after Nebulization

[0370] Select LNP-1 and LNP-8 to do example, collect after vibrating mesh nebulizer (Aerogen-Solo) atomization, detect the encapsulation efficiency of lipid nanoparticle composition, results such as Table 8 and Fig. 7.After atomizer atomization, LNP-8, i.e. the lipid nanoparticle composition corresponding to compound II-1, is affected by atomization shear force, and structure is destroyed, and encapsulation efficiency significantly decreases. And LNP-1, LNP-9 and LNP-10 can tolerate the influence of atomization shear force, and encapsulation efficiency remains consistent before and after atomization. Therefore, the lipid nanoparticle composition corresponding to compound I-1, compound I-8 and compound I-10 can be applicable to atomization inhalation and is used for pulmonary drug delivery.

[0371] Table 9

[0372] Test Example 5: Evaluation of mRNA protein expression in intratracheal delivery of lipid nanoparticle compositions

[0373] According to the preparation and detection methods of Example 6, lipid nanoparticle compositions were mixed and formulated with the molar percentages in Table 10 and characterized. The prepared lipid nanoparticles are shown in Tables 10 and 11.

[0374] Female C57 BL / 6 mice aged 6-8 weeks (purchased from Shanghai Southern Model Organisms Science Co., Ltd.) were intratracheally administered with a lipid nanoparticle composition encapsulating human hepatocyte growth factor (NCBI: NM_000601.6) at a dose of 0.5 mg / kg. 24 hours after administration, the mice were dissected and the lungs were removed to detect the protein expression of stem cell growth factor. The average protein expression was calculated for at least 3 biological replicates per group of lipid nanoparticle compositions. The relative protein expression of LNPs 11-17 was calculated, with the average protein expression of LNP-11 as the unit of 1. Taking LNP-12 as an example, the relative protein expression of LNP-12 = LNP-12 protein expression / LNP-11 protein expression.

[0375] Table 10

[0376] Table 11

[0377] Table 12

[0378] As shown in Table 12 and Figure 8, the protein expression levels of the lipid nanoparticle compositions containing compound I-1 were higher than those of the lipid nanoparticle compositions corresponding to compound II-1. Among them, the protein expression levels of the lipid nanoparticle compositions corresponding to LNP-12 and LNP-15 were the highest.

[0379] Test Example 6: Evaluation of mRNA delivery efficiency and tissue distribution of lipid nanoparticle compositions via tail vein injection in vivo

[0380] 6-8 week old female C57 BL / 6 mice (purchased from Shanghai Southern Model Organisms Science Co., Ltd.) were injected with a lipid nanoparticle composition encapsulating mRNA expressing luciferase (Luciferase, GenBank: MN728548.1) via the tail vein at a dose of 0.5 mg / kg. 6 hours after administration, each mouse was intraperitoneally injected with luciferase substrate (D-luciferin potassium salt abs42075819), and the lungs and livers of the mice were dissected. Fluorescence images were taken of the tissues using an IVIS small animal optical in vivo imaging instrument (PerkinElme), and the fluorescence intensity of each tissue was counted. The level of fluorescence intensity represents the level of luciferase protein expression, which reflects the efficiency of the lipid nanoparticle composition in delivering mRNA in vivo and its distribution in different organs and tissues. The average fluorescence intensity was calculated for at least 3 biological replicates of the lipid nanoparticles corresponding to each compound. The data are shown in Table 13 and Figure 9. The fluorescence intensity in Figure 9 is the fluorescence intensity corresponding to each tissue of the mouse captured and counted by the IVIS small animal optical in vivo imaging instrument. The fluorescence intensity represents the expression level of luciferase protein, which indicates the efficiency of mRNA delivery and tissue distribution of the lipid nanoparticle composition in vivo.

[0381] Using LNP-4 as a control, the mRNA delivery efficiency and tissue distribution of lipid nanoparticle compositions corresponding to the composition LNP-3 containing compound I-1 and compound II-1, the composition LNP-9 containing compound I-8 and compound II-1, and the composition LNP-10 containing compound I-10 and compound II-1 were detected after tail vein injection in mice.

[0382] Table 13

[0383] Conclusion: As shown in Table 13 and Figure 9, the fluorescence intensity of the lipid nanoparticle composition LNP-3 composed of compound I-1 has the highest fluorescence intensity in the lungs, that is, the expression of luciferase protein in the lungs is the highest, which is better than LNP-4, LNP-9 and LNP-10. The fluorescence intensity of the lung is divided by the fluorescence intensity of the liver to obtain the lung / liver ratio of tissue distribution. Among them, LNP-3 has the highest expression, followed by LNP-9, which is better than LNP-4 and LNP-10. That is, LNP containing compound I-1 and compound I-8 has higher expression in the lungs.

Claims

1. A lipid nanoparticle composition comprising: 1) a compound represented by formula I or a salt thereof, or 2) a compound represented by formula I or a salt thereof, and a compound represented by formula II or a salt thereof, in, L a and L b Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; H a , H b and H c Each independently selected from C 1-10 Alkylene; R a and R b Each independently selected from C 1-24 Alkyl and C 2-24 alkenyl; R c Selected from NR c1 R c2 and N + R c3 R c4 R c5 ; R c1 and R c2 are each independently selected from hydrogen and C 1-6 alkyl; R c3 , R c4 and R c5 Each independently selected from C 1-6 alkyl; L 1 and L 2 Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, and L 1 and L 2 At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; H 1 , H 2 and H 3 Each independently selected from C 1-10 Alkylene; R 1 and R 2 Each independently selected from C 1-24 Alkyl and C 2-24 alkenyl; R 3 Selected from hydroxyl groups.

2. The lipid nanoparticle composition according to claim 1, wherein L a and L b Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; L 1 and L 2 Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; Preferably, L a and L b At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; More preferably, L a and L b are each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-, and L a and L b At least one of them is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- or -C(O)O(CH2)2O-; L 1 and L 2 are each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH(CH3)C(O)O-, -OC(O)CH(CH3)O-, -OC(CH3)2C(O)O-, -OC(O)C(CH3)2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, -C(O)O(CH2)2O-, -O(CH2)4OC(O)- and -C(O)O(CH2)4O-, and L 1 and L 2 At least one of the following is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH(CH3)C(O)O-, -OC(O)CH(CH3)O-, -OC(CH3)2C(O)O-, -OC(O)C(CH3)2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)-, -C(O)O(CH2)2O-, -O(CH2)4OC(O)- and -C(O)O(CH2)4O-.

3. The lipid nanoparticle composition according to claim 1 or 2, wherein H a Selected from straight chain or branched C 1-10 Alkylene; H b Selected from straight chain or branched C 1-10 Alkylene; H c Selected from straight chain or branched C 1-10 Alkylene; H 1 Selected from straight or branched chain C 1-10 Alkylene; H 2 Selected from straight chain or branched C 1-10 Alkylene; H 3 Selected from straight or branched chain C 1-10 Alkylene; Preferably, H a Selected from straight or branched chain C 3-8 Alkylene; H b Selected from straight or branched chain C 3-8 Alkylene; H c Selected from straight or branched chain C 1-6 Alkylene; H 1 Selected from straight or branched chain C 3-8 Alkylene; H 2 Selected from straight or branched chain C 3-8 Alkylene; H 3 Selected from straight or branched chain C 1-6 Alkylene; More preferably, H a is selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-; H b is selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-; H c is selected from -CH2-, -(CH2)2-, -(CH2)3- and -(CH2)4-; H 1 is selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-; H 2 is selected from -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7- and -(CH2)8-; H 3 Selected from -CH2-, -(CH2)2-, -(CH2)3- and -(CH2)4-.

4. The lipid nanoparticle composition according to any one of claims 1 to 3, wherein R a and R b Each independently selected from a linear or branched C 1-24 Alkyl; R 1 and R 2 Each independently selected from a linear or branched C 1-24 alkyl; Preferably, R a and R b Each independently selected from a linear or branched C 4-18 Alkyl; R 1 and R 2 Each independently selected from a linear or branched C 4-18 alkyl; More preferably, R a Selected from straight chain C 4-18 Alkyl, R b Selected from branched C 4-18 Alkyl, or R a Selected from branched C 4-18 Alkyl, R b Selected from branched C 4-18 alkyl; R 1 Selected from straight chain C 4-18 Alkyl, R 2 Selected from branched C 4-18 Alkyl, or R 1 Selected from branched C 4-18 Alkyl, R 2 Selected from branched C 4-18 alkyl; More preferably, R a , R b , R 1 and R 2 Each independently selected from:

5. The lipid nanoparticle composition according to any one of claims 1 to 4, wherein The compound represented by formula I or its salt is a compound represented by formula Ia or its salt, Among them, L a , L b , H a , H b , H c , R a and R b As defined in any one of claims 1 to 4.

6. The lipid nanoparticle composition according to any one of claims 1 to 5, wherein The compound represented by formula I or its salt is selected from the compounds represented by formula IA, IB, IC, ID, IE and IF or their salts, Among them, H c , R a , R b and R c As defined in any one of claims 1 to 5; R d1 , R d2 , R d3 , R d4 , R d5 , R d6 , R e1 , R e2 , R e3 , R e4 , R e5 , R e6 , R f1 , R f2 , R f3 , R f4 , R f5 , R f6 , R g1 , R g2 , R g3 , R g4 , R g5 , R g6 , R h3 , R h4 , R h5 , R h6 , R i3 , R i4 , R i5 and R i6 are each independently selected from hydrogen and C 1-6 alkyl; n1, n2, n3, n4, n5, n6, n8, n9, n11, n12, n14 and n15 are each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; n7, n10, n13 and n16 are each independently selected from 0, 1, 2, 3, 4, 5 and 6.

7. The lipid nanoparticle composition according to any one of claims 1 to 6, wherein The compound represented by formula I is selected from the following structures, And / or, the compound represented by formula II is selected from compound II-1, 8. The lipid nanoparticle composition according to any one of claims 1 to 7, wherein The molar percentage of the compound represented by formula I or its salt in the lipid nanoparticle composition is 10% to 80%, preferably 10% to 60%; And / or, the molar percentage of the compound represented by formula II or its salt in the lipid nanoparticle composition is 5% to 50%, preferably 10% to 45%.

9. A lipid nanoparticle composition comprising: 1) a primary amine lipid; or, 2) a primary amine lipid, and a compound of formula II or a salt thereof according to any one of claims 1 to 7, in, L 1 , L 2 , H 1 , H 2 , H 3 , R 1 , R 2 and R 3 As defined in any one of claims 1 to 4; Preferably, the primary amine lipid is a compound represented by formula Ia according to claim 5 or a salt thereof, Among them, L a , L b , H a , H b , H c , R a and R b As defined in any one of claims 1 to 4.

10. The lipid nanoparticle composition according to claim 9, wherein The primary amine lipid accounts for 10% to 80% by mole of the lipid nanoparticle composition, preferably 10% to 60%; And / or, the molar percentage of the compound represented by formula II or its salt in the lipid nanoparticle composition is 5% to 50%, preferably 10% to 45%.

11. The lipid nanoparticle composition according to any one of claims 1 to 10, wherein The lipid nanoparticle composition comprises a phospholipid; Preferably, the phospholipid is selected from DGTS, DOPE and DSPC, and / or the phospholipid accounts for 1% to 25% by mole of the lipid nanoparticle composition; More preferably, DSPC, and / or, the phospholipids account for 1% to 15% by mole of the lipid nanoparticle composition.

12. The lipid nanoparticle composition according to any one of claims 1 to 11, wherein The lipid nanoparticle composition further comprises a steroid; Preferably, the steroid is selected from cholesterol, and / or the molar percentage of the steroid in the lipid nanoparticle composition is 10% to 50%; More preferably, the steroid accounts for 10% to 45% by mole of the lipid nanoparticle composition.

13. The lipid nanoparticle composition according to any one of claims 1 to 12, wherein The lipid nanoparticle composition further comprises a PEGylated lipid; Preferably, the PEGylated lipid is selected from PEGylated diacylglycerol, and / or the PEGylated lipid accounts for 0.5% to 10% by mole of the lipid nanoparticle composition; More preferably, the PEGylated lipid is selected from DMG-PEG, and / or the PEGylated lipid accounts for 0.5% to 5% by mole of the lipid nanoparticle composition.

14. The lipid nanoparticle composition according to any one of claims 1 to 13, wherein The composition preferentially delivers the therapeutic agent to a target organ selected from the group consisting of: lung, heart, liver, spleen, or stomach; Preferably, the target organ is the lung.

15. The lipid nanoparticle composition according to any one of claims 1-14 further comprises a therapeutic agent selected from the group consisting of small molecules, proteins and nucleic acids; Preferably, the therapeutic agent is selected from nucleic acids; More preferably, the nucleic acid is selected from mRNA.

16. A compound represented by formula III or a salt thereof, in, L A and L B Each independently selected from -O-, -C(O)-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, and L A and L B At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, where C 1-6 The alkylene group is not n-propylene; H a , H b , H c , R a , R b and R c As defined in any one of claims 1 to 7.

17. The compound or salt thereof according to claim 16, wherein L A and L B Each independently selected from -OC(O)-, -C(O)O-, -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-, where C 1-6 The alkylene group is not n-propylene; Preferably, L A and L B At least one selected from -OC 1-6 Alkylene-C(O)O-, -OC(O)-C 1-6 Alkylene-O-, -OC 1-6 Alkylene-OC(O)- and -C(O)OC 1-6 Alkylene-O-; More preferably, L A and L B are each independently selected from -OC(O)-, -C(O)O-, -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- and -C(O)O(CH2)2O-, and L A and L B At least one of them is selected from -OCH2C(O)O-, -OC(O)CH2O-, -OCH2OC(O)-, -C(O)OCH2O-, -O(CH2)2OC(O)- or -C(O)O(CH2)2O-.

18. The compound or salt thereof according to claim 16 or 17, wherein The compound represented by formula III or its salt is selected from the compounds represented by formula IIIA, IIIB, IIIC and IIID or their salts, Among them, H c , R a , R b and R c As defined in any one of claims 1 to 7; R d3 , R d4 , R d5 , R d6 , R e3 , R e4 , R e5 , R e6 , R f3 , R f4 , R f5 , R f6 , R g3 , R g4 , R g5 , R g6 , R h3 , R h4 , R h5 , R h6 , R i3 , R i4 , R i5 , R i6 As defined in claim 6; n5, n6, n7, n8, n9, n10, n11, n12, n13, n14, n15 and n16 are as defined in claim 6; Preferably, the compound represented by formula III or its salt is a compound or its salt having the following structure:

19. A pharmaceutical composition comprising the lipid nanoparticle composition according to any one of claims 1 to 15 or the compound or salt thereof according to any one of claims 16 to 18, and a pharmaceutically acceptable excipient.

20. Use of a lipid nanoparticle composition according to any one of claims 1 to 15, a compound or salt thereof according to any one of claims 16 to 18, or a pharmaceutical composition according to claim 19 in the preparation of a medicament for preventing and / or treating cancer, infection, autoimmune disease, neurodegenerative disease and inflammation.

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