Amino lipid compound as well as preparation method, composition and application thereof

Abstract

The invention discloses an amino lipid compound as well as a preparation method, a composition and application thereof. In particular to an amino lipid compound represented by a formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, use thereof for preparing lipid nanoparticles for delivering active ingredients, and a composition containing the amino lipid compound, especially lipid nanoparticles, and use thereof. # imgabs0 #

Description

Amino lipid compound, preparation method, composition and application thereof Technical Field

[0001] The present disclosure relates to amino lipid compounds that can be used to prepare lipid nanoparticles for delivering active ingredients and methods for preparing the same. The present disclosure also relates to compositions containing the amino lipid compounds, especially lipid nanoparticles, and uses thereof. Background Art

[0002] Gene medicine is a method of delivering genes with specific genetic information to target cells through artificial means, expressing target proteins or interfering with or regulating the expression of related genes to achieve clinical therapeutic effects. However, naked nucleic acids are difficult to introduce directly into cells, and gene delivery requires the help of external forces or carriers. However, differences in gene structure and cell structure require lipid particles of different properties to match. Therefore, it is necessary to develop different delivery vehicles or adjuvants, especially amino lipid compounds that can be used to deliver nucleic acid active ingredients and related preparation methods and applications to meet the delivery needs of gene medicines.

[0003] Summary of the Invention

[0004] One aspect of the present disclosure provides an amino lipid compound represented by formula (I):

[0005] wherein R1, R2, R3, R4, R5, Z1, Z2, Z3, Z4, Z5, Z6, Z7, Z8, A1, A2, A3, A4, A5, A6 and A7 are each as defined below.

[0006] Another aspect of the present disclosure provides a method for preparing the amino lipid compound.

[0007] Another aspect of the present disclosure provides use of the amino lipid compound for preparing a carrier for an active ingredient.

[0008] Another aspect of the present disclosure provides lipid nanoparticles containing the amino lipid compound.

[0009] Another aspect of the present disclosure provides a composition containing the amino lipid compound.

[0010] Another aspect of the present disclosure provides use of the amino lipid compound, lipid nanoparticle or composition in preparing a medicament.

[0011] Another aspect of the present disclosure provides use of the amino lipid compound, lipid nanoparticle or composition in the preparation of a medicament for nucleic acid transfer. DETAILED DESCRIPTION

[0012] definition

[0013] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. Reference to the technology used herein refers to the technology commonly understood in the art, including those changes in technology or equivalent technology substitutions that are obvious to those skilled in the art. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present disclosure.

[0014] As used herein, the terms "comprises," "comprising," "having," "containing," or "involving," and variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0015] The term "hydrocarbyl" as used herein means the group remaining after an aliphatic hydrocarbon loses one hydrogen atom, including straight-chain or branched, saturated or unsaturated hydrocarbon groups. Hydrocarbyl groups include alkyl, alkenyl and alkynyl groups. Preferably, the hydrocarbon group has 1 to 24 carbon atoms (C1-C 24 hydrocarbon radicals), for example, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms (C1, C2, C3, ... C 17 、C 18 、C 19 or C 20 Examples of hydrocarbon groups include, but are not limited to, C1-C 20 Hydrocarbon, C1-C 18 Hydrocarbon, C1-C 16 Hydrocarbon, C1-C 12 Hydrocarbon, C1-C 10 Hydrocarbon, C1-C8 hydrocarbon, C1-C7 hydrocarbon, C1-C6 hydrocarbon, C1-C4 hydrocarbon, C1-C3 hydrocarbon, C1-C2 hydrocarbon, C2-C8 hydrocarbon, C2-C4 hydrocarbon, C4-C8 hydrocarbon, C4-C9 hydrocarbon, C5-C8 hydrocarbon, C1-C4 hydrocarbon, C2-C8 hydrocarbon, C3 hydrocarbon, C4 hydrocarbon, C5 hydrocarbon, C6 hydrocarbon, C7 hydrocarbon and C8 hydrocarbon.

[0016] The term "alkylene group" used herein means a divalent group remaining after the above-mentioned alkyl group further loses one hydrogen atom.

[0017] The term "alkyl" as used herein is a straight-chain or branched saturated monovalent hydrocarbon group. Preferably, the alkyl group has 1 to 24 carbon atoms (C1-C 24 alkyl), for example having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms (C1, C2, C3, ... C17 、C 18 、C 19 or C 20 Examples of alkyl groups include, but are not limited to, C1-C 20 Alkyl, C1-C 18 Alkyl, C1-C 16 Alkyl, C1-C 12 Alkyl, C1-C 10 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, C2-C8 alkyl, C2-C4 alkyl, C4-C8 alkyl, C4-C9 alkyl, C5-C8 alkyl, C1-C4 alkyl, C2-C8 alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl and tridec-7-yl.

[0018] The term "alkylene" used herein refers to a divalent group remaining after the above-mentioned alkyl group further loses one hydrogen atom.

[0019] The term "alkenyl" as used herein is a linear or branched monovalent hydrocarbon group containing one or more double bonds (C=C). Preferably, the alkenyl group has 2 to 24 carbon atoms (C2-C 24 alkenyl), for example, having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms (C2, C3, ... C 17 、C 18 、C 19 or C 20 Alkenyl) and has 1, 2, 3, 4 or more double bonds. Alkenyl includes but is not limited to C2-C 20 Alkenyl, C2-C 18 Alkenyl, C2-C 16 Alkenyl, C2-C 12 Alkenyl, C2-C 10In some preferred embodiments, the alkenyl group comprises a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a C-C alkenyl, a

[0020] The term "alkenylene" used herein refers to a divalent group remaining after the above alkenyl group further loses one hydrogen atom.

[0021] The term "alkynyl" as used herein is a linear or branched monovalent hydrocarbon radical containing one or more triple bonds (C≡C). Preferably, the alkynyl radical has 2 to 24 carbon atoms (C2-C 24 alkynyl), for example having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms (C2, C3, ... C 17 、C 18 、C 19 or C 20 Alkynyl) and have 1, 2, 3, 4 or more triple bonds. Alkynyl includes but is not limited to C2-C 20 Alkynyl, C2-C 18 Alkynyl, C2-C 16 Alkynyl, C2-C 12 Alkynyl, C2-C 10 In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds. In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds. In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds. In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds. In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds. In some preferred embodiments, the alkynyl group comprises 1, 2, 3, 4 or more triple bonds.

[0022] The term "alkynylene group" used herein refers to a divalent group remaining after the above-mentioned alkynyl group further loses one hydrogen atom.

[0023] As used herein, the terms "cycloalkyl", "cycloalkylene" and "hydrocarbon ring" refer to saturated (i.e., "cycloalkyl" and "cycloalkylene") or unsaturated (i.e., having one or more double bonds (cycloalkenyl) and / or triple bonds (cycloalkynyl) within the ring) monocyclic or polycyclic hydrocarbon rings having, for example, 3-10 (suitably 3-8, more suitably 3-6, such as 5-6 or 5-7) ring carbon atoms, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclohexenyl, and the like.

[0024] The term "cycloalkyl" as used herein refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., a monocyclic ring such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or a bicyclic ring including spirocyclic, fused or bridged systems such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, decalinyl, etc.). The cycloalkyl group has, for example, 3 to 10, for example, 3-7, 5-6 or 5-7 carbon atoms.

[0025] As used herein, the term "heterocycle", "heterocyclyl" or "heterocyclylene" means a saturated or unsaturated cyclic group having ring heteroatoms selected from N, O and S. Preferably, the heterocycle is an optionally substituted 4 to 10 membered heterocycle having 1, 2, 3, 4, 5 or 6 ring heteroatoms selected from N, O and S. More preferably, the heterocycle is an optionally substituted 4 to 7 membered saturated heterocycle having 1, 2, 3 or 4 ring heteroatoms selected from N, O and S, more preferably an optionally substituted 5 to 7 membered (e.g., 5 to 6 membered) saturated heterocycle having 1, 2 or 3 ring heteroatoms selected from N, O and S. Examples of heterocycles include, but are not limited to, azetidine, oxetanyl, tetrahydrofuran, pyrrolidine, imidazolidine, pyrazolidine, tetrahydropyran, piperidine, morpholine, thiomorpholine, piperazine, preferably pyrrolidine, piperidine, piperazine and morpholine. The heterocycle may be optionally substituted by one or more substituents, as defined below for "optionally substituted".

[0026] As used herein, the term "aryl" refers to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π electron system. For example, as used herein, the term "C 6-14 "Aryl" means an aromatic group containing 6 to 14 (eg 6 to 12) carbon atoms, such as phenyl or naphthyl. Aryl is optionally substituted by 1 or more (eg 1 to 3) suitable substituents.

[0027] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) aromatic group having a conjugated π electron system, the ring atoms of which are composed of carbon atoms and at least one heteroatom, for example, having 5 to 14 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) ring atoms, wherein there are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 carbon atoms and 1, 2, 3, 4, or 5 identical or different heteroatoms independently selected from N, O, S, and S(O). One or more ring carbon atoms in the heteroaryl can be replaced by C(O). The heteroaryl can be benzo-fused. The heteroaryl can be optionally substituted by one or more suitable substituents.

[0028] As used herein, the term "optionally substituted" means that one or more hydrogen atoms attached to an atom or group are independently unsubstituted or substituted with one or more (e.g., 1, 2, 3, or 4) substituents. The substituents are independently selected from the group consisting of deuterium (D), halogen, -OH, mercapto, cyano, -CD3, C1-C6 alkyl (preferably C1-C3 alkyl), C2-C6 alkenyl, C2-C6 alkynyl, cycloalkyl (preferably C3-C8 cycloalkyl), aryl, heterocyclyl (preferably 3-8 membered heterocyclyl), heteroaryl, aryl C1-C6 alkyl-, heteroaryl C1-C6 alkyl, C1-C6 haloalkyl, -OC1-C6 alkyl (preferably -OC1-C3 alkyl), -OC2-C6 alkenyl, OC1-C6 alkylphenyl, C1-C6 alkyl-OH (preferably C1-C4 alkyl-OH), C1-C6 alkyl-SH, C1-C6 alkyl-O-C1-C6 alkyl, OC1-C6 haloalkyl, NH2, C1-C6 alkyl-NH2 (preferably C1-C3 alkyl-NH2), -N(C1-C6 alkyl)2 (preferably -N(C1-C3 alkyl)2), -NH(C1-C6 alkyl) (preferably -NH(C1-C3 alkyl)), -N(C1-C6 alkyl)(C1-C6 alkylphenyl), -NH(C1-C6 alkylphenyl), nitro, -C(O)-OH, -C(O)OC1-C6 alkyl (preferably -C(O)OC1-C3 alkyl), -CONRiRii (wherein Ri and Rii are H, D and C1-C6 alkyl, preferably C1-C3 alkyl), -NHC(O)(C1-C6 alkyl), -NHC(O)(phenyl), -N(C1-C6 alkyl) C(O)(C1-C6-alkyl), -N(C1-C6-alkyl)C(O)(phenyl), -C(O)C1-C6-alkyl, -C(O)heteroaryl (preferably -C(O)-5-7-membered heteroaryl), -C(O)C1-C6-alkylphenyl, -C(O)C1-C6-haloalkyl, -OC(O)C1-C6-alkyl (preferably -OC(O)C1-C3-alkyl), -S(O)2-C1-C6-alkyl, -S(O)-C1-C6-alkyl, -S(O)2-phenyl, -S(O)2-C1-C6-haloalkyl, -S(O)2NH2, -S(O)2NH(C1-C6-alkyl), -S(O)2NH(phenyl), -NHS(O)2(C1-C6-alkyl), -NHS(O)2(phenyl) and -NHS(O)2(C1-C6-haloalkyl),wherein each of the alkyl, cycloalkyl, phenyl, aryl, heterocyclyl and heteroaryl groups is optionally further substituted by one or more substituents selected from the group consisting of halogen, -OH, -NH2, cycloalkyl, 3-8 membered heterocyclyl, C1-C4 alkyl, C1-C4 haloalkyl-, -OC1-C4 alkyl, -C1-C4 alkyl-OH, -C1-C4 alkyl-O-C1-C4 alkyl, -OC1-C4 haloalkyl, cyano, nitro, -C(O)-OH, -C(O)OC1-C6 alkyl, -CON(C1-C6 -C1-C6 alkyl), -CONH2, -NHC(O)(C1-C6 alkyl), -NH(C1-C6 alkyl)C(O)(C1-C6 alkyl), -SO2(C1-C6 alkyl), -SO2(phenyl), -SO2(C1-C6 haloalkyl), -SO2NH2, -SO2NH(C1-C6 alkyl), -SO2NH(phenyl), -NHSO2(C1-C6 alkyl), -NHSO2(phenyl) and -NHSO2(C1-C6 haloalkyl). When an atom or group is substituted with multiple substituents, the multiple substituents may be the same or different.

[0029] In certain embodiments, the substituents can be independently selected from, but are not limited to, halogen (e.g., chlorine, bromine, fluorine, or iodine), deuterium (D), tritium (T), carboxylic acid (e.g., -C(=O)OH), oxygen (e.g., =O), sulfur (e.g., =S), hydroxyl (e.g., -OH), ester (e.g., -C(=O)ORiii or -OC(=O)Riii), aldehyde (e.g., -C(=O)H), carbonyl (e.g., -C(=O)Riii, or represented by C=O), acyl halide (e.g., -C(=O)X, wherein X is selected from bromine, fluorine, chlorine, or iodine), carbonate (e.g., -OC(=O)ORiii), alkoxy (e.g., -ORiii), acetal (e.g., -C(ORiii)2Riii, wherein each ORiii is the same or different alkoxy), phosphate (e.g., P(=O)4 3- ), sulfhydryl (e.g. -SH), sulfoxide (e.g. -S(=O)Riii), sulfinic acid (e.g. -S(=O)OH), sulfonic acid (e.g. -S(=O)2OH), thialdehyde (e.g. -C(=S)H), sulfate (e.g. S(=O)4 2-), sulfonyl (e.g., -S(=O)2Riii), sulfinyl (e.g., -S(=O)Riii), amide (e.g., -C(=O)N(Riii)2 or -N(Riii)C(=O)Riii), azido (e.g., -N3), nitro (e.g., -NO2), cyano (e.g., -CN), isocyano (e.g., -NC), acyloxy (e.g., -OC(=O)Riii), amino (e.g., -N(Riii)2, -N(Riii)H or -NH2), carbamoyl (e.g., -OC(=O)N(Riii)2, -OC(=O)Riii), )N(Riii)H or -OC(=O)NH2), sulfonamide (e.g., -S(=O)2N(Riii)2, -S(=O)2N(Riii)H, -S(=O)2NH2, -N(Riii)S(=O)2Riii, -N(H)S(=O)2Riii, -N(Riii)S(=O)2H or -N(H)S(=O)2H), alkyl, alkenyl, alkynyl, cycloalkyl (e.g., cycloalkyl, cycloalkenyl or cycloalkynyl), heterocycloalkyl (e.g., heterocycloalkyl containing one or more heteroatoms selected from S, N and O, or ... multiple heterocycles selected from S, N and O heteroatoms), aryl (e.g., phenyl, or fused ring), heteroaryl (e.g., 8-10 membered bicyclic heteroaryl containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur), -C(=O)SRiii, -C(=N-CN)N(Riii)2, -C(=NO-CH3)N(Riii)2, -C(=N-SO2-NH2)N(Riii)2, -C(=CH-NO2)N(Riii)2, -OC(=O)N(Riii)2, -CHN(Riii)N(Riii)2, - C(=O)N(Riii)ORiii、-N(Riii)2C(=O)ORiii、-OP(=O)(ORiii)2、-P(=O)(ORiii)2、-N(ORiii)C(=O)Riii、-N(ORiii)S(=O)2Riii、-N(ORiii)C(=O)ORiii、-N(ORiii)C(=O)N(Riii)2、-N(ORiii)C(=S)N(Riii)2、-N(ORiii)C(NRiii)N(Riii)2、-N(ORiii)C(CHRiii)N(Riii)2。In any of the foregoing, Riii is hydrogen as defined herein, or alkyl, or alkenyl, or alkynyl, or heteroalkyl, or heteroalkenyl, or heteroalkynyl. In some embodiments, Riii is hydrogen as defined herein, or C1-C 12 Alkyl, or C1-C 12 Alkenyl, or C1-C 12 Alkynyl, or C1-C 12 Heteroalkyl, or C1-C12 Heteroalkenyl, or C1-C 12 Heteroalkynyl.

[0030] In certain embodiments, the substituent itself can be further substituted by, for example, one or more substituents as defined herein. For example, a C1-C6 alkyl group as a substituent can be further substituted by one or more substituents as described herein.

[0031] In certain embodiments, the hydrocarbyl, hydrocarbylene, alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, cycloalkyl, cycloalkylene, hydrocarbyl ring, cycloalkyl, heterocycle, heterocyclyl, heterocyclylene, aryl, and heteroaryl groups described herein may be optionally substituted with one or more substituents.

[0032] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0033] As used herein, the groups "C(=O)" and "(C=O)" are used interchangeably to indicate For example, the group -(C=O)O- represents

[0034] As used herein, the term "compound" encompasses all compounds that have been isotopically labeled by replacing one or more atoms with atoms having a different atomic weight or mass number. "Isotopes" refer to atoms having the same atomic number but different mass numbers due to the number of neutrons in their nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0035] The numerical ranges described herein should be understood to encompass any and all subranges contained therein. For example, the range "1 to 10" should be understood to include not only the explicitly stated values ​​of 1 and 10, but also any individual value (e.g., 2, 3, 4, 5, 6, 7, 8, and 9) and subranges (e.g., 1 to 2, 1.5 to 2.5, 1 to 3, 1.5 to 3.5, 2.5 to 4, 3 to 4.5, etc.) within the range of 1 to 10. This principle also applies to ranges that use only one value as a minimum or maximum value.

[0036] As used herein, "pharmaceutically acceptable salts" refer to acid addition salts or base addition salts of the disclosed compounds that retain the biological effectiveness and properties of the disclosed compounds and, generally, are not biologically or otherwise undesirable. In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0037] Pharmaceutically acceptable acid addition salts can be formed with inorganic and / or organic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, decanoic acid, hexanoic acid, octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactose disulfonic acid, 1,2-di ... Acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid and undecylenic acid, etc.

[0038] Pharmaceutically acceptable base addition salts can be formed with inorganic bases and / or organic bases and the compounds of the present disclosure. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts and aluminum salts, etc. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, denanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0039] The compounds of the present disclosure or their pharmaceutically acceptable salts may contain one or more asymmetric centers and thus may produce enantiomers, diastereomers and other stereoisomeric forms, such as amino acids, which can be defined as (R)- or (S)-, or as (D)- or (L)-, based on absolute stereochemistry. The present invention is intended to include all of these possible isomers, as well as racemic and optically pure forms thereof. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques such as chromatography and fractional crystallization. Conventional techniques for preparing / isolating single enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.

[0040] As used herein, the term "stereoisomer" refers to a compound composed of the same atoms bonded by the same bonds but having different three-dimensional structures that are not interconvertible. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A mixture of a pair of enantiomers in any proportion is called a "racemic" mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0041] "Stereoisomers" may also include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compounds described herein are isolated as E or Z isomers. In other embodiments, the compounds described herein are mixtures of E and Z isomers.

[0042] "Tautomers" refers to isomeric forms of a compound that are in equilibrium with each other. The concentration of the isomeric forms will depend on the environment in which the compound is found, and may differ depending on, for example, whether the compound is a solid or in organic or aqueous solution.

[0043] Amino lipid compounds

[0044] In one aspect, the present disclosure provides an amino lipid compound represented by the following formula (I):

[0045] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0046] in:

[0047] Z1, Z2, Z5, Z6, Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS- or a bond;

[0048] Z3 is -C(=O)- or a bond;

[0049] Z4 is -O-, -CH(OH)-;

[0050] A1, A2, A3, A4, A5, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond;

[0051] R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle;

[0052] R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; and

[0053] R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle;

[0054] Preferably, A5, Z5 and Z6 are bonds.

[0055] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein A1 and A2 are each independently C1-C8 alkylene or a bond, such as a C1-C6 alkylene or a bond, or a C1-C4 alkylene or a bond. In some embodiments, A1 and A2 are each independently C1-C8 alkylene, C2-C8 alkenylene or C2-C8 alkynylene. In some embodiments, A1 and A2 are each independently C1-C8 alkylene, such as a C1-C6 alkylene, C1-C4 alkylene, C2-C4 alkylene or C1-C3 alkylene. In some embodiments, A1 and A2 are both bonds.

[0056] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein A3 is C1-C 12 In some embodiments, A3 is a linear C1-C 12 In some embodiments, A3 is a straight chain C1-C 12 Alkylene, C2-C 12Alkenylene or C2-C 12 In some embodiments, A3 is a linear C1-C5 alkylene, such as a linear C1, C2, C3, C4 or C5 alkylene. In some embodiments, A3 is -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0057] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein A4 is C1-C 12 In some embodiments, A4 is a linear C1-C 12 In some embodiments, A4 is a straight chain C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 In some embodiments, A4 is a linear C1-C7 alkylene, such as a linear C1, C2, C3, C4, C5, C6, or C7 alkylene. In some embodiments, A4 is a linear C1-C3 alkylene, such as a linear C1, C2, or C3 alkylene. In some embodiments, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0058] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein A5 is a C1-C8 alkylene group or a bond, such as a C1-C6 alkylene group or a bond, a C1-C4 alkylene group or a bond, or a C1-C3 alkylene group or a bond. In some embodiments, A5 is a C1-C8 alkylene group, a C2-C8 alkenylene group, or a C2-C8 alkynylene group. In some embodiments, A5 is a C1-C8 alkylene group, such as a C1-C6 alkylene group, a C1-C4 alkylene group, a C2-C4 alkylene group, or a C1-C3 alkylene group. In some embodiments, A5 is a bond.

[0059] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein A6 and A7 are each independently C1-C 12 In some embodiments, A6 and A7 are each independently a linear C1-C 12 In some embodiments, A6 and A7 are each independently a linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 In some embodiments, A6 and A7 are each independently a linear C1-C 12Alkylene, such as straight-chain C2-C 12 Alkylene, C4-C 10 Alkylene, C6-C8 alkylene, C6-C 10 In some embodiments, A6 and A7 are each independently a linear C6-C8 alkylene group. 10 Alkylene, such as linear C6, C7, C8, C9 or C 10 In some embodiments, A6 and A7 are each independently -(CH2)6-, -(CH2)7-, -(CH2)8-, or -(CH2)9-.

[0060] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1 and R2 are each independently: H; C1-C 18 Hydrocarbon groups, such as C1-C 16 Hydrocarbon, C1-C 12 Hydrocarbon, C1-C 10 alkyl, C1-C8 hydrocarbon, C1-C6 hydrocarbon or C1-C4 hydrocarbon; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or a 3- to 7-membered heterocycle, such as a 4- to 6-membered heterocycle; or, R1 and R2, together with the nitrogen atom to which they are attached, form a 4- to 7-membered heterocycle, such as a 5- to 6-membered heterocycle.

[0061] In some embodiments, R1 and R2 are each independently: H; C1-C 18 Alkylene, C2-C 18 Alkenylene or C2-C 18 In some embodiments, R1 and R2 are each independently C1-C 18 alkyl, such as C1-C4 alkyl; C3-C6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; phenyl; or a 5- to 6-membered heterocyclic ring. In some embodiments, R1 and R2, together with the nitrogen atom to which they are attached, form a 5- to 6-membered heterocyclic ring, such as pyrrolidine, piperidine, piperazine or morpholine.

[0062] In some embodiments, R1 and R2 are each independently C1-C4 alkyl, such as C1, C2, C3 or C4 alkyl. In some embodiments, R1 and R2 are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl. In some embodiments, R1 and R2 are each independently C1-C3 alkyl, such as C1, C2 or C3 alkyl. In some embodiments, R1 and R2 are each independently methyl, ethyl or n-propyl.

[0063] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R3 is: H; C1-C 18 Hydrocarbon groups, such as C1-C 16 Hydrocarbon, C2-C 12 Hydrocarbon, C4-C 10 alkyl, or C4-C8 alkyl; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or 3 to 7 membered heterocyclic, such as 4 to 6 membered heterocyclic. In some embodiments, R3 is C1-C 18 Alkyl, C2-C 18 Alkenyl or C2-C 18 In some embodiments, R3 is C1-C 18 Alkyl groups, such as C2-C 10 In some embodiments, R3 is a linear C4-C8 alkyl group, such as a linear C4, C5, C6, C7, or C8 alkyl group. In some embodiments, R3 is n-butyl, n-pentyl, n-hexyl, n-heptyl, or n-octyl group.

[0064] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z1, Z2, Z5, and Z6 are each independently -C(=O)-, -O(C=O)-, -O-, or a bond. In some embodiments, Z1, Z2, Z5, and Z6 are each independently a bond.

[0065] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z3 is -C(=O)-.

[0066] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-, or a bond.

[0067] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein one of Z7 or Z8 is -(C=O)O-, -O(C=O)-, or a bond.

[0068] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is -(C=O)O-. In some embodiments, the (C=O) in Z7 is connected to A6, and the (C=O) in Z8 is connected to A7.

[0069] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is -O(C=O)-. In some embodiments, (C=O) in Z7 is connected to R4, and (C=O) in Z8 is connected to R5.

[0070] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein one of Z7 or Z8 is a bond. In some embodiments, Z7 is a bond and Z8 is -(C=O)O-. In some embodiments, the (C=O) in Z8 is connected to A7. In some embodiments, Z7 is a bond and Z8 is -O(C=O)-. In some embodiments, the (C=O) in Z8 is connected to R5. In some embodiments, Z8 is a bond and Z7 is -(C=O)O-. In some embodiments, the (C=O) in Z7 is connected to A6. In some embodiments, Z8 is a bond and Z7 is -O(C=O)-. In some embodiments, the (C=O) in Z7 is connected to R4.

[0071] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is a bond.

[0072] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently: C1-C 20 alkyl; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or 3 to 7 membered heterocyclic, such as 4 to 6 membered heterocyclic. In some embodiments, R4 and R5 are each independently: C1-C 20 C3-C6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; phenyl; or a 5- to 6-membered heterocyclic ring.

[0073] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently C1-C 20 Hydrocarbon group.

[0074] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently C1-C 20 alkyl.

[0075] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently a branched C3-C 20 Alkyl groups, such as branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 In some embodiments, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 In some embodiments, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13 In some embodiments, R4 is a branched alkyl group with branching occurring at the α, β, or γ position relative to Z7, for example: R5 is a branched alkyl group in which the branching occurs at the α, β or γ position relative to Z8, for example: In some embodiments, R4 is a branched alkyl group with branching occurring at the α or β position relative to Z7; and R5 is a branched alkyl group with branching occurring at the α or β position relative to Z8.

[0076] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures:

[0077] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures:

[0078] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently

[0079] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein:

[0080] Z3 is -C(=O)-; and

[0081] A4 is C1-C 12 In some embodiments, A4 is a linear C1-C 12 Alkylene, such as straight chain C1-C 10 In some embodiments, A4 is -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0082] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z4 is -O-.

[0083] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z4 is -CH(OH)-.

[0084] In another aspect, the present disclosure provides amino lipid compounds represented by the following formula (II) and formula (III):

[0085] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0086] in:

[0087] Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS-, or a bond;

[0088] A3, A4, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond;

[0089] R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle;

[0090] R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; and

[0091] R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle.

[0092] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-, or a bond. In some embodiments, each of Z7 and Z8 is -(C=O)O-. In some embodiments, the (C=O) in Z7 is connected to A6, and the (C=O) in Z8 is connected to A7. In some embodiments, each of Z7 and Z8 is -O(C=O)-. In some embodiments, the (C=O) in Z7 is connected to R4, and the (C=O) in Z8 is connected to R5.

[0093] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein A3, A4, A6 and A7 are each independently C1-C 12 In some embodiments, A3, A4, A6 and A7 are each independently C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 Alkylene.

[0094] In some embodiments, A3 is a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12In some embodiments, A3 is a linear C1, C2, C3, C4, or C5 alkylene. In some embodiments, A3 is a linear C2, C3, or C4 alkylene, such as -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0095] In some embodiments, A4 is a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 In some embodiments, A4 is a linear C1, C2, C3, or C4 alkylene group, such as -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0096] In some embodiments, A6 and A7 are each independently a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 In some embodiments, A6 and A7 are each independently a linear C6, C7, C8, C9 or C 10 Alkylene, for example -(CH2)6-, -(CH2)7-, -(CH2)8- or -(CH2)9-.

[0097] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R1 and R2 are each independently C1-C 18 In some embodiments, R1 and R2 are each independently C1-C6 alkyl. In some embodiments, R1 and R2 are each independently methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R1 and R2 are each independently C1-C4 alkyl, such as C1, C2, C3, or C4 alkyl. In some embodiments, R1 and R2 are each independently methyl.

[0098] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclic ring. In some embodiments, R1 and R2, together with the nitrogen atom to which they are attached, form a 5-6 membered heterocyclic ring.

[0099] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R3 is C1-C18 In some embodiments, R3 is C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 In some embodiments, R3 is a linear C2, C3, C4, C5, C6, C7, C8, C9 or C 10 In some embodiments, R3 is a linear C4, C5, C6, C7, or C8 alkyl group, such as n-butyl, n-pentyl, n-hexyl, n-heptyl, or n-octyl.

[0100] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R4 and R5 are each independently C1-C 20 Hydrocarbon group.

[0101] In some embodiments, R4 and R5 are each independently C1-C 20 alkyl.

[0102] In some embodiments, R4 and R5 are each independently a branched C3-C 20 Alkyl groups, such as branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 In some embodiments, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 In some embodiments, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13Alkyl. In some embodiments, R4 is a branched alkyl group with branching occurring at the ɑ, β, or γ position relative to Z7; R5 is a branched alkyl group with branching occurring at the ɑ, β, or γ position relative to Z8. In some embodiments, R4 is a branched alkyl group with branching occurring at the ɑ or β position relative to Z7; R5 is a branched alkyl group with branching occurring at the ɑ or β position relative to Z8. In some embodiments, R4 and R5 are each independently one of the following structures:

[0103] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R4 and R5 are each independently one of the following structures:

[0104] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein R4 and R5 are each independently

[0105] In some embodiments, the present disclosure provides amino lipid compounds of formula (II) and formula (III) as described above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, wherein:

[0106] Z7 and Z8 are each independently -(C=O)O- or -O(C=O)-;

[0107] A3 is a straight-chain C1-C5 alkylene group;

[0108] A4 is a straight-chain C1-C4 alkylene group;

[0109] A6 and A7 are each independently C5-C 11 alkylene;

[0110] R1 and R2 are each independently C1-C4 alkyl;

[0111] R3 is a straight chain C2-C 10 Alkyl; and

[0112] R4 and R5 are each independently C1-C 24 alkyl.

[0113] In some embodiments, each of Z7 and Z8 is -(C=O)O-, and the (C=O) in Z7 is connected to A6, and the (C=O) in Z8 is connected to A7. In some embodiments, each of Z7 and Z8 is -O(C=O)-, and the (C=O) in Z7 is connected to R4, and the (C=O) in Z8 is connected to R5.

[0114] In some embodiments, A3 is a linear C1-C4 alkylene. In some embodiments, A3 is a linear C2-C4 alkylene, such as a linear C2, C3 or C4 alkylene. In some embodiments, A3 is -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0115] In some embodiments, A4 is a linear C1-C3 alkylene, such as a linear C1, C2 or C3 alkylene. In some embodiments, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0116] In some embodiments, A6 and A7 are each independently a linear C5-C 11 In some embodiments, A6 and A7 are each independently a linear C6-C 10 Alkylene, such as linear C6, C7, C8, C9 or C 10 In some embodiments, A6 and A7 are each independently a linear C7, C8, or C9 alkylene group, such as -(CH2)7-, -(CH2)8-, or -(CH2)9-.

[0117] In some embodiments, R1 and R2 are each independently C1, C2, C3 or C4 alkyl. In some embodiments, R1 and R2 are each independently methyl, ethyl, n-propyl or isopropyl. In some embodiments, R1 and R2 are each independently methyl.

[0118] In some embodiments, R3 is a linear C4-C8 alkyl, such as a linear C4, C5, C6, C7 or C8 alkyl. In some embodiments, R3 is n-butyl, n-pentyl, n-hexyl, n-heptyl or n-octyl.

[0119] In some embodiments, R4 and R5 are each independently a branched C3-C 20 Alkyl groups, such as branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 In some embodiments, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 In some embodiments, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13 Alkyl. In some embodiments, R4 is a branched alkyl group with branching occurring at the ɑ, β, or γ position relative to Z7; R5 is a branched alkyl group with branching occurring at the ɑ, β, or γ position relative to Z8. In some embodiments, R4 is a branched alkyl group with branching occurring at the ɑ or β position relative to Z7; R5 is a branched alkyl group with branching occurring at the ɑ or β position relative to Z8.

[0120] In some embodiments, R4 and R5 are each independently one of the following structures:

[0121] In some embodiments, R4 and R5 are each independently one of the following structures:

[0122] In some embodiments, R4 and R5 are each independently

[0123] In various embodiments, the present disclosure provides an amino lipid compound of formula (I), (II) or (III) as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the amino lipid compound has one of the structures shown in Table 1 below.

[0124] Table 1

[0125] The amino lipid compounds disclosed herein all have hydrophobic properties due to the presence of long non-polar residues, and simultaneously have hydrophilic properties due to the amino groups. Due to this amphiphilic property, the amino lipid compounds disclosed herein can be used to form lipid nanoparticles, such as lipid bilayers, micelles, liposomes, and the like.

[0126] In the context of the present disclosure, the term "lipid nanoparticles" refers to nanosized substances obtained by placing amino lipid compounds in aqueous solution. These particles are particularly lipid nanoparticles, lipid bilayer vesicles (liposomes), multilamellar vesicles or micelles.

[0127] In some preferred embodiments, the lipid nanoparticles are liposomes containing the amino lipid compounds of the present disclosure.In the context of the present disclosure, liposomes are microvesicles composed of a bilayer of lipid amphiphilic molecules enclosing an aqueous compartment.

[0128] Liposome formation is not a spontaneous process. Lipid vesicles first form when lipids are placed in water, forming a bilayer or a series of bilayers, each separated by water molecules. Liposomes can be formed by treating lipid vesicles in water with ultrasound.

[0129] For the purposes of this disclosure, the term "lipid bilayer" refers to a thin membrane formed by two layers of lipid molecules. The term "micelle" refers to an aggregate of surfactant molecules dispersed in a liquid colloid. Typical micelles in aqueous solutions form aggregates upon contact with water, with the hydrophilic head regions sequestering the hydrophobic single tail region at the center of the micelle.

[0130] In one aspect, the present disclosure provides a use of the amino lipid compound of the present disclosure for preparing a carrier of an active ingredient. In some embodiments, the carrier is in the form of a lipid nanoparticle, such as a lipid bilayer, micelle, or liposome.

[0131] lipid nanoparticles

[0132] In another aspect, the present disclosure provides lipid nanoparticles containing the amino lipid compounds of the present disclosure.

[0133] In some embodiments, the lipid nanoparticle further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0134] In some embodiments, the lipid nanoparticles further contain one or more of a helper lipid, a structural lipid, and a PEG-lipid (polyethylene glycol-lipid).

[0135] In some further embodiments, the lipid nanoparticle further contains the helper lipid, the structural lipid, and the PEG-lipid.

[0136] In some embodiments, the lipid nanoparticles comprise the following amounts (molar percentages) of the amino lipid compound, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid, and the PEG-lipid: about 25.0%-75.0%, for example, about 25.0%-28.0%, 28.0%-32.0%, 32.0%-35.0%, 35.0%-40.0%, 40.0%-5 ... 0.0%-42.0%, 42.0%-45.0%, 45.0%-48.0%, 48.0%-55.0%, 55.0%-65.0%, 65.0%-75.0%, 45.0%-46.3%, 46.3%-48.0%, 48.0%-49.5%, 49.5%-50.0%, 50.0%-55.0% or 60.0%-65.0%.

[0137] In some embodiments, the lipid nanoparticles comprise the following amount (molar percentage) of the helper lipid, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid, and the PEG-lipid: about 5.0%-45.0%, for example, about 5.0%-10.0%, 10.0%-16.0%, 16.0%-25.0%, 25.0%-33.5%, 33.5%-37.0%, 37.0%-38.0%, 39.0%-40.0%, 40.0%-45 ... 7.0%-40.0%, 40.0%-42.0%, 42.0%-45.0%, 5.0%-9.0%, 9.0%-9.4%, 9.4%-10.0%, 10.0%-10.5%, 10.5%-11.0%, 11.0%-15.0%, 15.0%-16.0%, 16.0%-18.0%, 18.0%-20.0% or 20.0%-25.0%.

[0138] In some embodiments, the lipid nanoparticles comprise the following amount (molar percentage) of the structural lipid, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid, and the PEG-lipid: about 0.0%-50.0%, for example, about 0.0%-10.0%, 10.0%-15.5%, 15.5%-22.5%, 22.5%-35.0%, 35.0%-36.5%, 36.5%-39.5%, 39.5%-40.5%, 40.5%-41.5%, 41.5%-45.0%, 45.0%-46.5%. 5%, 46.5%-50.0%, 15.5%-18.5%, 18.5%-22.5%, 22.5%-23.5%, 23.5%-28.5%, 28.5%-33.5%, 33.5%-35.0%, 36.5%-38.0%, 38.0%-38.5%, 38.5%-39.0%, 39.0%-39.5%, 41.5%-42.5%, 42.5%-42.7%, 42.7%-43.0%, 43.0%-43.5%, 43.5%-45.0% or 46.5%-48.5%. In some embodiments, the lipid nanoparticle comprises the structural lipid in an amount (molar percentage) of about 50.0%-55.0%, such as about 50.0%-51.5%, 51.5%-53.5%, or 53.5%-55.0%.

[0139] In some embodiments, the lipid nanoparticles comprise the PEG-lipid in an amount (molar percentage) of about 0.5%-5.0%, for example, about 0.5%-1.0%, 1.0%-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, 4.5%-5.0%, 1.5%-1.6% or 1.6%-2.0%, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid and the PEG-lipid.

[0140] In some embodiments as described above, the helper lipid is a phospholipid. The phospholipid is generally semi-synthetic, but may also be of natural origin or chemically modified. The phospholipid includes, but is not limited to, DSPC (distearoylphosphatidylcholine), DOPE (dioleoylphosphatidylethanolamine), DOPC (dioleoylphosphatidylcholine), DOPS (dioleoylphosphatidylserine), DSPG (1,2-dioctadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol)), DPPG (dipalmitoylphosphatidylglycerol), DPPC (dipalmitoylphosphatidylcholine), DGTS (1,2-dipalmitoyl-sn-glycero-3-O-4'-(N,N,N-trimethyl)homoserine), lysophospholipids, and the like. Preferably, the helper lipid is one or more selected from DSPC, DOPE, DOPC, and DOPS. In some embodiments, the helper lipid is DSPC and / or DOPE.

[0141] In some embodiments, the structured lipid is a sterol substance, including but not limited to cholesterol, cholesterol ester, sterol hormones, sterol vitamins, bile acid, cholesterol, ergosterol, β-sitosterol and oxidized cholesterol derivatives. Preferably, the structured lipid is at least one selected from cholesterol, cholesterol ester, sterol hormones, sterol vitamins and bile acids. In some embodiments, the structured lipid is cholesterol, preferably high-purity cholesterol, especially injection-grade high-purity cholesterol, such as CHO-HP (produced by AVT).

[0142] As used herein, the term PEG-lipid (polyethylene glycol-lipid) is a conjugate of polyethylene glycol and a lipid structure. Preferably, the PEG-lipid is selected from PEG-DMG and PEG-distearoylphosphatidylethanolamine (PEG-DSPE), preferably PEG-DMG. Preferably, the PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristyl glyceride. Preferably, the average molecular weight of the PEG is about 2000 to 5000, preferably about 2000, such as PEG2000-DMG.

[0143] In some embodiments as described above, in the lipid nanoparticles, the molar ratio of the amino lipid compound of the present disclosure: helper lipid: structural lipid: PEG-lipid is about 45:11:41.5:2.5, or 42.0:10.5:45.0:2.5, or 42.0:16.0:39.5:2.5, or 40.0:16.0:41.5:2.5, or 40.0:18.0:39.5:2.5, or 35.0: 16.0:46.5:2.5, or 35.0:25.0:36.5:3.5, or 28.0:33.5:35.0:3.5, or 32.0:37.0:40.5:0.5, or 35.0:40.0:22.5:2.5, or 40.0:42.0:15.5:2.5, or 45:10:42.5:2.5, or 40.0:20.0:38.5:1.5, or 45.0:15.0:3 8.5:1.5, or 55.0:5.0:38.5:1.5, or 60.0:5.0:33.5:1.5, or 45.0:20.0:33.5:1.5, or 50.0:20.0:28.5:1.5, or 55.0:20.0:23.5:1.5, or 60.0:20.0:18.5:1.5, or 40.0:15.0:43.5:1.5, or 50.0:15.0:33.5 : 1.5, or 55.0:15.0:28.5:1.5, or 60.0:15.0:23.5:1.5, or 40.0:10.0:48.5:1.5, or 45.0:10.0:43.5:1.5, or 55.0:10.0:33.5:1.5, or 40.0:5.0:53.5:1.5, or 45.0:5.0:48.5:1.5, or 50.0:5.0:43.5:1.5. In some such embodiments, the helper lipid is DOPE and the structural lipid is CHO-HP.

[0144] In other embodiments as described above, in the lipid nanoparticles, the molar ratio of the amino lipid compound of the present disclosure: helper lipid: structural lipid: PEG-lipid is about 48.0:10.0:40.5:1.5, or 50.0:10.0:38.5:1.5, or 50.0:9.0:38.0:3.0, or 49.5:10.0:39.0:1.5, or 46.3:9.4:42.7:1.6, or 45.0:10.0:38.5:1.5. :9.0:43.0:3.0, or 45.0:11.0:41.5:2.5, or 42.0:10.5:45.0:2.5, or 42.0:16.0:39.5:2.5, or 40.0:16.0:41.5:2.5, or 40.0:18.0:39.5:2.5, or 35.0:40.0:22.5:2.5, or 40.0:20.0:38.5:1.5, or 45.0:15. 0:38.5:1.5, or 55.0:5.0:38.5:1.5, or 60.0:5.0:33.5:1.5, or 45.0:20.0:33.5:1.5, or 50.0:20.0:28.5:1.5, or 55.0:20.0:23.5:1.5, or 60.0:20.0:18.5:1.5, or 40.0:15.0:43.5:1.5, or 50.0:15.0:33. : 1.5, or 55.0:15.0:28.5:1.5, or 60.0:15.0:23.5:1.5, or 40.0:10.0:48.5:1.5, or 45.0:10.0:43.5:1.5, or 55.0:10.0:33.5:1.5, or 40.0:5.0:53.5:1.5, or 45.0:5.0:48.5:1.5, or 50.0:5.0:43.5:1.5. In some such embodiments, the helper lipid is DSPC and the structural lipid is CHO-HP.

[0145] In some embodiments, the lipid nanoparticles have the amino lipid compound, helper lipid, structural lipid and PEG-lipid of the present disclosure in the mole percentages (%) shown in No. 1-12, No. 13-24 in Table 2 below, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid and the PEG-lipid:

[0146] Table 2

[0147] In some embodiments, the lipid nanoparticles have the amino lipid compound, helper lipid, structural lipid and PEG-lipid of the present disclosure in the mole percentages (%) shown in No. 25-42 in Table 3 below, based on the total amount of the amino lipid compound, the helper lipid, the structural lipid and the PEG-lipid:

[0148] Table 3

[0149] As described above, the lipid nanoparticles of the present disclosure can be used as carriers of active ingredients.

[0150] In some embodiments, the active ingredient includes a therapeutic and / or prophylactic agent.

[0151] The term "therapeutic agent" or "prophylactic agent" refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or prophylactic effect and / or elicits a desired biological and / or pharmacological effect.

[0152] An "effective amount" or "therapeutically effective amount" refers to an amount of an amino lipid compound of the present disclosure or a lipid nanoparticle comprising the amino lipid compound of the present disclosure that is sufficient to achieve treatment in a mammal (preferably a human) when administered to the mammal (preferably a human). The amount of lipid nanoparticles of the present disclosure that constitutes a "therapeutically effective amount" will depend on the amino lipid compound, the condition and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one of ordinary skill in the art based on their own knowledge and this disclosure.

[0153] In some embodiments, the active ingredient is a pharmaceutical active ingredient, including but not limited to anti-tumor agents, antibiotics, immunomodulators, anti-inflammatory agents, agents acting on the central nervous system, antigens or fragments thereof, proteins, peptides, polypeptides, polypeptoids, vaccines and small molecules, and mixtures thereof. Preferably, the pharmaceutical active ingredient is a biologically active ingredient.

[0154] Within the scope of the present disclosure, a biologically active ingredient is a substance that has a biological effect when introduced into a cell or host, for example, by stimulating an immune response or an inflammatory response, by exerting enzymatic activity, or by complementing a mutation, etc. The biologically active ingredients include, but are not limited to, nucleic acids, antigens or fragments thereof, proteins, peptides, polypeptides, polypeptides, antibodies, vaccines, and small molecules, and mixtures thereof.

[0155] When lipid nanoparticles encapsulate active ingredients in their internal aqueous space, they can be referred to as "lipid nanoparticle drugs."

[0156] In the context of the present disclosure, the term "cell" is a general term and includes cultures of single cells, tissues, organs, insect cells, avian cells, fish cells, amphibian cells, mammalian cells, primary cells, continuous cell lines, stem cells, and / or genetically engineered cells (e.g., recombinant cells expressing heterologous polypeptides or proteins). Recombinant cells include, for example, cells expressing heterologous polypeptides or proteins (e.g., growth factors or blood factors).

[0157] In some preferred embodiments, the bioactive component is a nucleic acid.

[0158] In some embodiments as described above, the lipid nanoparticles of the present disclosure further contain a nucleic acid.

[0159] In some embodiments, in the lipid nanoparticles, the mass ratio of the amino lipid compound of the present disclosure to the nucleic acid is about (5-30):1, such as about (5-10):1, (10-15):1, (15-20):1, (20-25):1 or (25-30):1, preferably about 10:1.

[0160] In some embodiments, the nucleic acid is selected from RNA, antisense oligonucleotide, DNA.

[0161] In some embodiments, the RNA is selected from messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single-stranded guide RNA (sgRNA), cas9 mRNA, or a mixture thereof.

[0162] In some embodiments, the messenger RNA (mRNA) encodes a polypeptide and / or protein of interest. This includes any naturally occurring or non-naturally occurring or otherwise modified polypeptide. In some embodiments, the polypeptide and / or protein encoded by the mRNA can have a therapeutic and / or preventive effect when expressed in a cell.

[0163] In some embodiments, the RNA is a siRNA that can selectively reduce the expression of a gene of interest or downregulate the expression of the gene. For example, the selection of the siRNA can silence the gene associated with a particular disease, disorder, or condition after the lipid nanoparticles comprising the siRNA are administered to a subject in need. The siRNA can comprise a sequence complementary to the mRNA sequence encoding a gene or protein of interest. In some embodiments, the siRNA can be an immunomodulatory siRNA.

[0164] In certain embodiments, the RNA is sgRNA and / or cas9 mRNA. sgRNA and / or cas9 mRNA can be used as gene editing tools. For example, the sgRNA-cas9 complex can affect the translation of mRNA of a cellular gene.

[0165] In some embodiments, the RNA is shRNA or its encoding vector or plasmid. shRNA can be produced inside the target cell after the appropriate construct is delivered to the nucleus. The construct and mechanism associated with shRNA are well known in the relevant art.

[0166] In some embodiments, the DNA is a plasmid.

[0167] In some embodiments, the lipid nanoparticles are used to transfer nucleic acids. In some embodiments, the lipid nanoparticles can be used, for example, for gene therapy, genetic vaccination, protein replacement therapy, antisense therapy, or therapy performed by interfering RNA.

[0168] Composition

[0169] In another aspect, the present disclosure provides a composition comprising the lipid nanoparticles as described above and a pharmaceutically acceptable carrier, diluent, or excipient.

[0170] In some embodiments, the composition further comprises a buffer. In some such embodiments, the buffer is selected from phosphate buffer and Tris buffer, preferably phosphate buffer. In some embodiments, the concentration of the buffer is about 5 mmol / L-30 mmol / L, preferably about 10 mmol / L. In some embodiments, the pH of the buffer is about 6-8, preferably about 7-8, and more preferably about 7-7.5.

[0171] In some embodiments, the composition further comprises a cryoprotectant. In some such embodiments, the cryoprotectant is selected from sucrose and trehalose, preferably sucrose. In some embodiments, the concentration of the cryoprotectant is about 50 mg / ml-100 mg / ml.

[0172] In some embodiments as described above, the composition further comprises a cryoprotectant. In some such embodiments, the cryoprotectant is selected from sucrose and trehalose, preferably sucrose. In some embodiments, the concentration of the cryoprotectant is about 50 mg / ml-100 mg / ml.

[0173] In some embodiments, the composition is a pharmaceutical composition.

[0174] use

[0175] The lipid nanoparticles of the present disclosure have excellent performance in encapsulating bioactive ingredients. Lipid nanoparticles containing bioactive ingredients can be used to deliver any of a variety of therapeutic agents into cells. The present disclosure includes the use of the lipid nanoparticles described above to deliver bioactive ingredients into cells. The present disclosure also provides a method for delivering a bioactive ingredient into a cell, the method comprising contacting a lipid nanoparticle of the present disclosure containing the bioactive ingredient with the cell.

[0176] The lipid nanoparticles of the present disclosure can be used to transfect multicellular tissues or organs. Therefore, the present disclosure also provides a method for transfecting cells, multicellular tissues or organs, the method comprising contacting the lipid nanoparticles of the present disclosure containing the nucleic acid with the cells, tissues or organs. This provides new therapeutic treatment possibilities for individuals.

[0177] In some embodiments, the cell is a mammalian cell; more preferably, the mammalian cell is in a mammal.

[0178] In some embodiments, the tissue or organ is selected from the group consisting of spleen, liver, kidney, lung, femur, eye tissue, vascular endothelium in blood vessels, lymphatic, and tumor tissue.

[0179] As used herein, a subject can be any mammal, preferably selected from mice, rats, pigs, cats, dogs, horses, goats, cows, monkeys, etc. In some preferred embodiments, the subject is a human.

[0180] In another aspect, the present disclosure provides a method for producing a polypeptide and / or protein of interest in a mammalian cell, the method comprising contacting the cell with a lipid nanoparticle comprising mRNA encoding the polypeptide and / or protein of interest, wherein after the cell is contacted with the lipid nanoparticle, the mRNA can be taken up into the cell and translated to produce the polypeptide and / or protein of interest.

[0181] In another aspect, the present disclosure provides the use of the amino lipid compound, lipid nanoparticle or composition of the present disclosure in the preparation of a drug. In another aspect, the present disclosure provides the lipid nanoparticle or composition of the present disclosure for use as a drug. In another aspect, the present disclosure provides a drug comprising the lipid nanoparticle or composition of the present disclosure. In some embodiments, the drug is used for the treatment and / or prevention of a disease. In some embodiments, the disease is selected from the group consisting of: rare diseases, infectious diseases, cancer, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases. In some embodiments, the drug is used for, for example, gene therapy, gene vaccination, protein replacement therapy, antisense therapy or treatment by interfering RNA. Preferably, the gene therapy can be used for the treatment of cancer and genetic diseases. In some embodiments, the cancer is one or more selected from lung cancer, gastric cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, blood cancer or prostate cancer. In some embodiments, the genetic disease is one or more selected from hemophilia, thalassemia, and Gaucher's disease. In some embodiments, the gene vaccine is preferably used to treat cancer, allergy, toxicity and pathogen infection. In some embodiments, the pathogen is one or more selected from viruses, bacteria or fungi.

[0182] In yet another aspect, the present disclosure provides a method of treating a disease or disorder in a mammal in need thereof, comprising administering to the mammal a therapeutically effective amount of the lipid nanoparticle according to the aforementioned method.

[0183] Preferably, the disease or condition is selected from the group consisting of rare diseases, infectious diseases, cancer, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases.

[0184] In another aspect, the disclosure provides amino lipid compounds, lipid nanoparticles or compositions for use in the preparation of a drug for nucleic acid transfer. In another aspect, the disclosure provides lipid nanoparticles or compositions of the disclosure for use as a drug for nucleic acid transfer. In another aspect, the disclosure provides a drug comprising lipid nanoparticles or compositions of the disclosure for nucleic acid transfer. In some embodiments, the nucleic acid is selected from RNA, antisense oligonucleotides, and DNA. In some embodiments, the RNA is selected from emissive RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA). In some embodiments, the DNA is a plasmid.

[0185] Preparation method

[0186] In another aspect, the present disclosure also provides a general synthetic method for preparing the amino lipid compound of formula (II) of the present disclosure, as shown below:

[0187] wherein A3, A4, A6, A7, R1, R2, R3, R4, R5, Z7 and Z8 each have the same meaning as defined above for the amino lipid compound of formula (I) or formula (II).

[0188] Specifically, the method includes:

[0189] Step 1: Reductive amination of a compound of formula DTN with a compound of formula di-N to obtain a compound of formula F; and

[0190] Step 2: Reacting the compound of formula F with the compound of formula G to obtain the amino lipid compound of formula (II).

[0191] In some embodiments, the reductive amination reaction in step 1 is carried out in an organic solvent (e.g., a C1-C3 alcohol, preferably ethanol). In some embodiments, the reductive amination reaction is carried out in the presence of a catalyst (e.g., palladium on carbon) under a hydrogen atmosphere. In some embodiments, the reductive amination reaction is carried out at an elevated pressure (e.g., about 2-8 or about 4-6 MPa) and an elevated temperature (e.g., about 50-90, or about 60-80° C.).

[0192] In some embodiments, the reaction described in Step 2 is performed in the presence of a base (eg, pyridine or DMAP).

[0193] In another aspect, the present disclosure also provides a general synthetic method for preparing the amino lipid compound of formula (III) of the present disclosure, as shown below:

[0194] wherein A3, A4, A6, A7, R1, R2, R3, R4, R5, Z7 and Z8 each have the same meaning as defined above for the compound of formula (I) or formula (III).

[0195] Specifically, the method includes:

[0196] Step 1: Reductive amination of a compound of formula DTN with a compound of formula di-N to obtain a compound of formula F; and

[0197] Step 2: Reacting the compound of formula F with the compound of formula H to obtain the amino lipid compound of formula (III).

[0198] In some embodiments, the reductive amination reaction in step 1 is carried out in an organic solvent (e.g., a C1-C3 alcohol, preferably ethanol). In some embodiments, the reductive amination reaction is carried out in the presence of a catalyst (e.g., palladium on carbon) under a hydrogen atmosphere. In some embodiments, the reductive amination reaction is carried out at an elevated pressure (e.g., about 2-8 or about 4-6 MPa) and an elevated temperature (e.g., about 50-90, or about 60-80° C.).

[0199] In another aspect, the lipid nanoparticles or compositions of the present invention can be prepared according to methods known in the art. For example, the method can include the following steps:

[0200] (1) Ingredients: preparing a suitable aqueous phase; and preparing an organic phase comprising the amino lipid compound of the present disclosure and optionally a helper lipid, a structural lipid and / or a PEG-lipid;

[0201] (2) Encapsulation: mixing appropriate amounts of the aqueous phase and the organic phase;

[0202] (3) Dialysis: Optionally, the mixture of step (2) is dialyzed;

[0203] (4) Sterilization: The product of step (3) is optionally sterilized, for example, by passing it through a sterilizing filter, such as a 0.22 μm microporous filter membrane.

[0204] In some embodiments, the lipid nanoparticles or compositions of the present disclosure containing nucleic acids, particularly mRNA, can be prepared by a method comprising the following steps:

[0205] (1) Ingredients: preparing an aqueous phase containing the nucleic acid; and preparing an organic phase (e.g., an ethanol phase) containing the amino lipid compound of the present disclosure and optionally a helper lipid, a structural lipid, and / or a PEG-lipid;

[0206] (2) Encapsulation: mixing appropriate amounts of the aqueous phase and the organic phase;

[0207] (3) Dialysis: Optionally, the mixture of step (2) is dialyzed;

[0208] (4) Sterilization: The product of step (3) is optionally sterilized, for example, by passing it through a sterilizing filter, such as a 0.22 μm microporous filter membrane.

[0209] The present disclosure also includes the following embodiments:

[0210] Embodiment 1. An amino lipid compound having a structure of formula (I):

[0211] or a pharmaceutically acceptable salt or stereoisomer thereof,

[0212] in:

[0213] Z1, Z2, Z5, Z6, Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS- or a bond;

[0214] Z3 is -C(=O)- or a bond;

[0215] Z4 is -O-, -CH(OH)-;

[0216] A1, A2, A3, A4, A5, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond;

[0217] R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle;

[0218] R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle;

[0219] R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle;

[0220] Preferably, A5, Z5 and Z6 are bonds.

[0221] Embodiment 2. The amino lipid compound of embodiment 1, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-, or a bond.

[0222] Embodiment 3. The amino lipid compound of embodiment 1, wherein one of Z7 or Z8 is -(C=O)O-, -O(C=O)-, or a bond.

[0223] Embodiment 4. The amino lipid compound according to embodiment 1, wherein each of Z7 and Z8 is -(C=O)O-.

[0224] Embodiment 5. The amino lipid compound of embodiment 1, wherein one of Z7 or Z8 is a bond.

[0225] Embodiment 6. The amino lipid compound of embodiment 1, wherein each of Z7 and Z8 is a bond.

[0226] Embodiment 7. The amino lipid compound according to embodiment 1, wherein R4 and R5 are each independently C1-C 20 Hydrocarbon group.

[0227] Embodiment 8. The amino lipid compound according to embodiment 1, wherein R4 and R5 are each independently C1-C 20 alkyl.

[0228] Embodiment 9. The amino lipid compound according to embodiment 1, wherein R4 and R5 are each independently a branched C3-C 20 alkyl.

[0229] Embodiment 10. The amino lipid compound of embodiment 1, wherein R4 and R5 are each independently:

[0230] Embodiment 11. An amino lipid compound having a structure of formula (II):

[0231] or a pharmaceutically acceptable salt or stereoisomer thereof,

[0232] in:

[0233] Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS-, or a bond;

[0234] A3, A4, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond;

[0235] R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle;

[0236] R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle;

[0237] R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle.

[0238] Embodiment 12. The amino lipid compound according to embodiment 11, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-.

[0239] Embodiment 13. The amino lipid compound according to embodiment 11, wherein A3, A4, A6 and A7 are each independently C1-C 12 Alkylene.

[0240] Embodiment 14. The amino lipid compound according to embodiment 11, wherein R1 and R2 are each independently C1-C 18 alkyl.

[0241] Embodiment 15. According to the amino lipid compound of embodiment 14, R1 and R2 are each independently a C1-C6 alkyl group.

[0242] Embodiment 16. According to the amino lipid compound of embodiment 14, R1 and R2 are each independently methyl, ethyl, n-propyl or isopropyl.

[0243] Embodiment 17. The amino lipid compound of embodiment 11, wherein R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclic ring.

[0244] Embodiment 18. The amino lipid compound of embodiment 17, wherein R1 and R2, together with the nitrogen atom to which they are attached, form a 5-6 membered heterocyclic ring.

[0245] Embodiment 19. The amino lipid compound according to embodiment 11, wherein R3 is C1-C 18 alkyl.

[0246] Embodiment 20. The amino lipid compound according to embodiment 11, wherein R4 and R5 are each independently C1-C 20 Hydrocarbon group.

[0247] Embodiment 21. The amino lipid compound according to embodiment 11, wherein R4 and R5 are each independently C1-C 20 alkyl.

[0248] Embodiment 22. The amino lipid compound according to embodiment 11, wherein R4 and R5 are each independently a branched C3-C 18 alkyl.

[0249] Embodiment 23. The amino lipid compound of embodiment 11, wherein R4 and R5 are each independently:

[0250] Embodiment 24. An amino lipid compound having a structure of formula (III):

[0251] or a pharmaceutically acceptable salt or stereoisomer thereof,

[0252] in:

[0253] Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS-, or a bond;

[0254] A3, A4, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond;

[0255] R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle;

[0256] R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle;

[0257] R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle.

[0258] Embodiment 25. The amino lipid compound according to embodiment 24, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-.

[0259] Embodiment 26. The amino lipid compound according to embodiment 24, wherein A3, A4, A6 and A7 are each independently C1-C 12 Alkylene.

[0260] Embodiment 27. The amino lipid compound according to embodiment 24, wherein R1 and R2 are each independently C1-C 18 alkyl.

[0261] Embodiment 28. According to the amino lipid compound of embodiment 27, R1 and R2 are each independently a C1-C6 alkyl group.

[0262] Embodiment 29. According to the amino lipid compound of embodiment 28, R1 and R2 are each independently methyl, ethyl, n-propyl or isopropyl.

[0263] Embodiment 30. The amino lipid compound of embodiment 24, wherein R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclic ring.

[0264] Embodiment 31. The amino lipid compound of embodiment 24, wherein R1 and R2, together with the nitrogen atom to which they are attached, form a 5-6 membered heterocyclic ring.

[0265] Embodiment 32. The amino lipid compound according to embodiment 24, wherein R3 is C1-C 18 alkyl.

[0266] Embodiment 33. The amino lipid compound according to embodiment 24, wherein R4 and R5 are each independently C1-C 20 Hydrocarbon group.

[0267] Embodiment 34. The amino lipid compound according to embodiment 24, wherein R4 and R5 are each independently C1-C 20 alkyl.

[0268] Embodiment 35. The amino lipid compound according to embodiment 24, wherein R4 and R5 are each independently a branched C3-C 18 alkyl.

[0269] Embodiment 36. The amino lipid compound of embodiment 24, wherein R4 and R5 are each independently:

[0270] Embodiment 37. A lipid nanoparticle comprising the amino lipid compound according to any one of embodiments 1 to 36, and optionally a pharmaceutically acceptable carrier, diluent or excipient.

[0271] Embodiment 38. The lipid nanoparticle of embodiment 37, further comprising one or more of a helper lipid, a structural lipid, and a PEG-lipid;

[0272] Furthermore, the lipid nanoparticles further contain the auxiliary lipid, the structural lipid and the PEG-lipid.

[0273] Embodiment 39. The lipid nanoparticle of embodiment 38, wherein the lipid nanoparticle comprises the amino lipid compound of any one of embodiments 1 to 36 in an amount (molar percentage) of about 25.0%-75.0%, such as about 25.0%-28.0%, 28.0%-32.0%, 32.0%-35.0%, 35.0%-40.0%, 40.0%-42.0%, 42.0%-45.0%, 45.0%-48.0%, 48.0%-55.0%, 55.0%-65.0%, or 65.0%-75.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0274] Embodiment 40. The lipid nanoparticle of any one of embodiments 38 or 39, wherein the lipid nanoparticle comprises the helper lipid in an amount (molar percentage) of about 5.0%-45.0%, such as about 5.0%-10.0%, 10.0%-16.0%, 16.0%-25.0%, 25.0%-33.5%, 33.5%-37.0%, 37.0%-40.0%, 40.0%-42.0%, or 42.0%-45.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0275] Embodiment 41. The lipid nanoparticle of any one of embodiments 38 to 40, wherein the lipid nanoparticle comprises the structural lipid in an amount (molar percentage) of about 0.0%-50.0%, such as about 0.0%-10.0%, 10.0%-15.5%, 15.5%-22.5%, 22.5%-35.0%, 35.0%-36.5%, 36.5%-39.5%, 39.5%-40.5%, 40.5%-41.5%, 41.5%-45.0%, 45.0%-46.5%, or 46.5%-50.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0276] Embodiment 42. The lipid nanoparticle of any one of embodiments 38 to 41, wherein the lipid nanoparticle comprises the PEG-lipid in an amount (molar percentage) of about 0.5%-5.0%, such as about 0.5%-1.0%, 1.0%-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, or 4.5%-5.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0277] Embodiment 43. A lipid nanoparticle according to any one of embodiments 38 to 42, wherein the helper lipid is one or more selected from DSPC, DOPE, DOPC, DOPS, DSPG, DPPG, DPPC, DGTS and lysophospholipids, preferably one or more selected from DSPC, DOPE, DOPC and DOPS, more preferably DSPC and / or DOPE.

[0278] Embodiment 44. The lipid nanoparticle of any one of embodiments 38 to 43, wherein the structural lipid is at least one selected from cholesterol, cholesterol esters, steroid hormones, steroid vitamins, bile acids, cholesterol, ergosterol, β-sitosterol, and oxidized cholesterol derivatives, preferably at least one selected from cholesterol, cholesterol esters, steroid hormones, steroid vitamins, and bile acids, more preferably cholesterol, even more preferably high-purity cholesterol, particularly injection-grade high-purity cholesterol, such as CHO-HP.

[0279] Embodiment 45. The lipid nanoparticle of any one of embodiments 38 to 44, wherein the PEG-lipid is selected from PEG-DMG and PEG-DSPE, preferably PEG-DMG;

[0280] Preferably, the PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristyl glycerol;

[0281] Preferably, the average molecular weight of the PEG is about 2000 to 5000, preferably about 2000.

[0282] Embodiment 46. The lipid nanoparticle of any one of embodiments 38 to 45, wherein the molar ratio of the amino lipid compound of any one of embodiments 1 to 36: helper lipid: structural lipid: PEG-lipid is about 45:11:41.5:2.5, or 42.0:10.5:45.0:2.5, or 42.0:16.0:39.5:2.5, or 40.0:16.0:4 1.5:2.5, or 40.0:18.0:39.5:2.5, or 35.0:16.0:46.5:2.5, or 35.0:25.0:36.5:3.5, or 28.0:33.5:35.0:3.5, or 32.0:37.0:40.5:0.5, or 35.0:40.0:22.5:2.5, or 40.0:42.0:15.5:2.5.

[0283] Embodiment 47. The lipid nanoparticle of embodiment 46, wherein the auxiliary lipid is DOPE and the structural lipid is CHO-HP.

[0284] Embodiment 48. The lipid nanoparticle of any one of embodiments 38 to 45, wherein the molar ratio of the amino lipid compound of any one of embodiments 1 to 36:helper lipid:structural lipid:PEG-lipid is about 48.0:10.0:40.5:1.5.

[0285] Embodiment 49. The lipid nanoparticle of embodiment 48, wherein the helper lipid is DSPC and the structural lipid is CHO-HP.

[0286] Embodiment 50. The lipid nanoparticle of any one of embodiments 37 to 49, further comprising a nucleic acid.

[0287] Embodiment 51. The lipid nanoparticle of embodiment 50, wherein the mass ratio of the amino lipid compound according to any one of embodiments 1 to 36 and the nucleic acid is about (5-30):1, such as about (5-10):1, (10-15):1, (15-20):1, (20-25):1 or (25-30):1, preferably about 10:1.

[0288] Embodiment 52. The lipid nanoparticle of embodiment 50 or 51, wherein the nucleic acid is selected from RNA, antisense oligonucleotide, DNA;

[0289] Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA);

[0290] Preferably, the DNA is a plasmid.

[0291] Embodiment 53. A composition comprising the amino lipid compound according to any one of embodiments 1 to 36 and a pharmaceutically acceptable carrier, diluent or excipient.

[0292] Embodiment 54. The composition of embodiment 53, further comprising one or more of a helper lipid, a structural lipid, and a PEG-lipid;

[0293] Furthermore, the composition further contains the helper lipid, the structural lipid and the PEG-lipid.

[0294] Embodiment 55. The composition of embodiment 54, wherein the composition comprises the amino lipid compound of any one of embodiments 1 to 36 in an amount (molar percentage) of about 25.0%-75.0%, such as about 25.0%-28.0%, 28.0%-32.0%, 32.0%-35.0%, 35.0%-40.0%, 40.0%-42.0%, 42.0%-45.0%, 45.0%-48.0%, 48.0%-55.0%, 55.0%-65.0%, or 65.0%-75.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipids, the structural lipids, and the PEG-lipids.

[0295] Embodiment 56. The composition of any one of embodiments 54 to 55, wherein the composition comprises the helper lipid in an amount (molar percentage) of about 5.0%-45.0%, such as about 5.0%-10.0%, 10.0%-16.0%, 16.0%-25.0%, 25.0%-33.5%, 33.5%-37.0%, 37.0%-40.0%, 40.0%-42.0%, or 42.0%-45.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0296] Embodiment 57. The composition of any one of embodiments 54 to 56, wherein the composition comprises the following amount (molar percentage) of the structural lipid, based on the total amount of the amino lipid compound according to any one of embodiments 1 to 36, the helper lipid, the structural lipid and the PEG-lipid: about 0.0%-50.0%, such as about 0.0%-10.0%, 10.0%-15.5%, 15.5%-22.5%, 22.5%-35.0%, 35.0%-36.5%, 36.5%-39.5%, 39.5%-40.5%, 40.5%-41.5%, 41.5%-45.0%, 45.0%-46.5%, or 46.5%-50.0%.

[0297] Embodiment 58. The composition of any one of embodiments 54 to 57, wherein the composition comprises the PEG-lipid in an amount (molar percentage) of about 0.5%-5.0%, such as about 0.5%-1.0%, 1.0%-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, or 4.5%-5.0%, based on the total amount of the amino lipid compound of any one of embodiments 1 to 36, the helper lipid, the structural lipid, and the PEG-lipid.

[0298] Embodiment 59. A composition according to any one of embodiments 54 to 58, wherein the helper lipid is selected from one or more of DSPC, DOPE, DOPC, DOPS, DSPG, DPPG, DPPC, DGTS and lysophospholipids, preferably one or more of DSPC, DOPE, DOPC and DOPS, more preferably DSPC and / or DOPE.

[0299] Embodiment 60. A composition according to any one of embodiments 54 to 58, wherein the structured lipid is at least one selected from cholesterol, cholesterol esters, steroid hormones, steroid vitamins, bile acids, cholesterol, ergosterol, β-sitosterol and oxidized cholesterol derivatives, preferably at least one selected from cholesterol, cholesterol esters, steroid hormones, steroid vitamins and bile acids, more preferably cholesterol, even more preferably high-purity cholesterol, in particular injection-grade high-purity cholesterol, such as CHO-HP.

[0300] Embodiment 61. The composition of any one of embodiments 54 to 58, wherein the PEG-lipid is selected from PEG-DMG and PEG-DSPE, preferably PEG-DMG;

[0301] Preferably, the PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristyl glycerol;

[0302] Preferably, the average molecular weight of the PEG is about 2000 to 5000, preferably about 2000.

[0303] Embodiment 62. The composition of any one of embodiments 54 to 61, wherein the molar ratio of the amino lipid compound of any one of embodiments 1 to 36: helper lipid: structural lipid: PEG-lipid is about 45:11:41.5:2.5, or 42.0:10.5:45.0:2.5, or 42.0:16.0:39.5:2.5, or 40.0:16.0:41 .5:2.5, or 40.0:18.0:39.5:2.5, or 35.0:16.0:46.5:2.5, or 35.0:25.0:36.5:3.5, or 28.0:33.5:35.0:3.5, or 32.0:37.0:40.5:0.5, or 35.0:40.0:22.5:2.5, or 40.0:42.0:15.5:2.5.

[0304] Embodiment 63. A composition according to embodiment 62, wherein the helper lipid is DOPE and the structural lipid is CHO-HP.

[0305] Embodiment 64. The composition of any one of embodiments 54 to 61, wherein the molar ratio of the amino lipid compound of any one of embodiments 1 to 36:helper lipid:structural lipid:PEG-lipid is about 48.0:10.0:40.5:1.5.

[0306] Embodiment 65. The composition of embodiment 64, wherein the helper lipid is DSPC and the structural lipid is CHO-HP.

[0307] Embodiment 66. The composition of any one of Embodiments 53 to 65, further comprising a nucleic acid.

[0308] Embodiment 67. A composition according to embodiment 66, wherein the mass ratio of the amino lipid compound according to any one of embodiments 1 to 36 and the nucleic acid is about (5-30):1, such as about (5-10):1, (10-15):1, (15-20):1, (20-25):1 or (25-30):1, preferably about 10:1.

[0309] Embodiment 68. The composition of embodiment 66 or 67, wherein the nucleic acid is selected from RNA, antisense oligonucleotide, DNA;

[0310] Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA);

[0311] Preferably, the DNA is a plasmid.

[0312] Embodiment 69. The composition of any one of embodiments 53 to 68, further comprising a buffer;

[0313] Preferably, the buffer is selected from phosphate buffer and Tris buffer, preferably phosphate buffer; and / or

[0314] Preferably, the concentration of the buffer is about 5 mmol / L-30 mmol / L, preferably about 10 mmol / L; and / or

[0315] Preferably, the pH of the buffer is about 6-8, preferably about 7-8, more preferably about 7-7.5.

[0316] Embodiment 70. The composition of any one of embodiments 53 to 68, further comprising a cryoprotectant;

[0317] Preferably, the cryoprotectant is selected from sucrose and trehalose, preferably sucrose; and / or

[0318] Preferably, the concentration of the cryoprotectant is about 50 mg / ml-100 mg / ml.

[0319] Embodiment 71. Use of the amino lipid compound according to any one of embodiments 1 to 36 for the preparation of a carrier for an active ingredient.

[0320] Embodiment 72. The use according to embodiment 71, wherein:

[0321] The carrier is a lipid nanoparticle according to any one of embodiments 37 to 52; and / or

[0322] The active ingredient is a pharmaceutical active ingredient, preferably a nucleic acid; and / or

[0323] Preferably, the nucleic acid is selected from RNA, antisense oligonucleotide, DNA;

[0324] Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA);

[0325] Preferably, the DNA is a plasmid.

[0326] Embodiment 73. Use of the amino lipid compound of any one of embodiments 1 to 36, the lipid nanoparticle of any one of embodiments 37 to 52, or the composition of any one of embodiments 53 to 70 in the preparation of a medicament.

[0327] Embodiment 74. The use according to embodiment 73, wherein the medicament is for gene therapy, genetic vaccination, protein replacement therapy, antisense therapy or treatment by interfering RNA.

[0328] Embodiment 75. The use according to embodiment 74, wherein the gene therapy is used for the treatment of cancer and genetic diseases.

[0329] Embodiment 76. The use according to embodiment 75, wherein the cancer is one or more selected from lung cancer, stomach cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, blood cancer or prostate cancer; and the genetic disease is one or more selected from hemophilia, thalassemia and Gaucher's disease.

[0330] Embodiment 77. The use according to embodiment 75, wherein the genetic vaccination is used to treat cancer, allergy, toxicity and pathogen infection.

[0331] Embodiment 78. The use according to embodiment 77, wherein the pathogen is one or more selected from viruses, bacteria or fungi.

[0332] Embodiment 79. Use of the amino lipid compound of any one of embodiments 1 to 36, the lipid nanoparticle of any one of embodiments 37 to 52, or the composition of any one of embodiments 53 to 70 in the preparation of a medicament for nucleic acid transfer.

[0333] Embodiment 80. The use according to embodiment 79, wherein the nucleic acid is selected from RNA, antisense oligonucleotide, DNA;

[0334] Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA);

[0335] Preferably, the DNA is a plasmid. Beneficial effects

[0336] The amino lipid compounds disclosed herein can form carriers with excellent bioactive ingredient encapsulation properties, such as lipid nanoparticles, which can be used to deliver bioactive ingredients, particularly poorly water-soluble drugs or easily degradable or degradable active ingredients (e.g., nucleic acids), thereby improving their bioavailability and efficacy, immunocompetence, transfection efficiency (for nucleic acids), safety, or preference for specific organs or tissues. In particular, lipid nanoparticles containing the amino lipid compounds disclosed herein have a significant preference for delivery to the spleen.

[0337] Lipid nanoparticles containing the amino lipid compounds disclosed herein have obvious delivery preferences for immune cells in different tissues and organs, including spleen, lymph, peripheral blood, bone marrow, lung, liver, etc., and immune cells include neutrophils, NK cells-NK T cells, macrophages-monocytes, dendritic cells, B cells, T cells, CD4 T cells, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0338] Figures 1A to 1F show the delivery efficiency of lipid nanoparticles containing different amino lipid compounds to immune cell populations in different tissues and organs, among which Figure 1A is neutrophils; Figure 1B is NK cells-NK T cells; Figure 1C is macrophages-monocytes; Figure 1D is dendritic cells; Figure 1E is B cells; and Figure 1F is CD4 T cells.

[0339] In order to make the purpose, technical solutions and advantages of the present disclosure more clear, the present disclosure is described below in conjunction with specific embodiments. The following embodiments are only used to illustrate the present disclosure, rather than to limit it.

[0340] Example

[0341] The following examples are provided for purposes of illustration and not limitation.

[0342] Experimental methods without specific conditions specified in the examples are generally conventional conditions, or conditions recommended by the raw material or product manufacturers; reagents without source specified are generally conventional reagents available through commercial channels.

[0343] The abbreviations used in the examples have the following meanings: Pd / C palladium / carbon; EA ethyl acetate; DCM dichloromethane; MPa megapascal; DMF N,N-dimethylformamide; EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; DMAP 4-dimethylaminopyridine; TEMPO 2,2,6,6-tetramethylpiperidinium N-oxide; NaDCC sodium dichloroisocyanurate; h hour; min minute.

[0344] Example 1. Synthesis of amino lipid compound 125

[0345] (1) Synthesis of compound 1522F-1

[0346] A 200ml autoclave was charged with 10.0g DTN-T (14.1mmol), 2.88g N,N-dimethylaminopropylamine, 0.5g Pd / C (10%), and 80ml of anhydrous ethanol. The autoclave was sealed and purged with nitrogen three times, then hydrogen three times, and then filled with hydrogen to a pressure of 4MPa. The temperature was then raised to 70°C. After reacting for 48 hours, the autoclave was filtered, the filter cake was washed with anhydrous ethanol, and the filtrate was concentrated to obtain 11g of a crude product. This crude product was chromatographed on a silica gel column using 9:1 EA:methanol as the eluent to obtain 9g of 1522F-1, for an 80% yield.

[0347] (2) Synthesis of Compound 125-G

[0348] To a 500mL two-necked flask, add 8.67g of sodium hydroxide and 195ml of DMF. Cool to 0°C under a nitrogen-protected descender, then slowly add 15.0g of 1,3-propylene glycol (197.1mmol). After reacting for 30 minutes, warm to room temperature, continue to react for 1 hour, and then cool to 0°C. Slowly add 35.7g of 1-bromopentane dropwise to the reaction solution. After reacting for 30 minutes, warm to room temperature, continue to react for 15 hours, add 100mL of water, extract with 100mL of EA, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The concentrate was chromatographed on a silica gel column using EA:n-hexane = 1:8 as the eluent to obtain 17.10g of 125G-1, with a yield of 59.3%.

[0349] To a 1000 mL round-bottom flask were added 17 g of 125G-1 (116.2 mmol), 100 mL of DCM, 0.54 g of TEMPO, 23.3 g of KHCO₃, and 1.2 g of NaBr. With stirring, 38.34 g of NaDCC aqueous solution was added. The reaction was stirred at room temperature for 15 h and then filtered. The organic phase was separated from the filtrate, and the aqueous phase was extracted twice with 100 mL of DCM. The combined organic phases were concentrated under reduced pressure, and the concentrate was chromatographed on a silica gel column using 1:8 EA:n-hexane as the eluent to afford 11 g of 125-G, a 64.7% yield.

[0350] 1 H NMR in(CDCl3)δ3.71-3.69(m,2H),3.47-3.44(m,2H),2.64-2.61(m,2H),1.57-1.54(m,2H),1.34-1.25(m,6H),0.89-0.87(m,3H).

[0351] (3) Synthesis of amino lipid compound 125

[0352] To a 100 ml round-bottom flask, 0.47 g EDCI, 0.02 g DMAP, and 30 ml DCM were added sequentially. After stirring for 10 min, a solution of 1522F-1 (1.3 g, 1.64 mmol) in DCM (3 ml), a solution of 125-G (0.34 g) in DCM (3 ml), and a solution of pyridine (0.2 g) in DCM (3 ml) were added dropwise. After stirring at room temperature for 15 h, the pH was adjusted to 3 with dilute hydrochloric acid, 30 ml of water was added, and the mixture was extracted with 30 ml of DCM. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 1.5 g of crude product. The crude product was chromatographed on a silica gel column using EA:n-hexane = 1.5:1 as the eluent to obtain amino lipid compound 125 (1.1 g) with a purity of 95.88% and a yield of 72%.

[0353] 1 H NMR(CDCl3)δ4.86(m,2H),3.74-3.72(m,2H),3.62(m,1H),3.43-3.42(m,2H),3.12(m,2H) ,2.60(m,2H),2.28-2.21(m,12H),1.71-1.44(m,20H),1.25(m,56H),0.87-0.86(m,15H).

[0354] LC-MS (ESI): (M+H) 936.4.

[0355] Example 2. Synthesis of amino lipid compound 120

[0356] According to the general synthesis method, amino lipid compound 120 (0.56 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.31 g 120-G with a purity of 90.85% and a yield of 37%.

[0357] 1 H NMR(CDCl3)δ4.86-4.84(m,2H),4.13-4.09(m,2H),3.55-3.46(m,3H),3.14-3.11(m, 2H),2.30-2.21(m,12H),1.74-1.44(m,20H),1.31-1.25(m,56H),0.87-0.85(m,15H).

[0358] LC-MS (ESI): (M+H) 922.4.

[0359] Example 3. Synthesis of amino lipid compound 130

[0360] According to the general synthetic method, amino lipid compound 130 (1.05 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.37 g 130-G with a purity of 92.04% and a yield of 67%.

[0361] 1 H NMR(CDCl3)δ4.87-4.85(m,2H),3.65(m,1H),3.45-3.43(m,2H),3.39-3.37(m,2H),3.12-3.11(m,2H),2.41 -2.37(m,2H),2.30-2.22(m,12H),1.93-1.89(m,2H),1.71-1.44(m,20H),1.25(m,56H),0.88-0.86(m,15H).

[0362] LC-MS (ESI): (M+H) 950.4.

[0363] Example 4. Synthesis of amino lipid compound 144

[0364] According to the general synthetic method, amino lipid compound 144 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.28 g of 144-G as a colorless oil with a purity of 95.67% and a yield of 68%.

[0365] 1 H NMR(CDCl3)δ4.87-4.85(m,2H),4.15-4.09(m,2H),3.51-3.47(m,3H),3.25-3.23(m,2H),2. 47-2.45(m,2H),2.28-2.25(m,10H),1.60-1.35(m,20H),1.25(m,52H),0.92-0.85(m,15H).

[0366] LC-MS (ESI): (M+H) 894.3.

[0367] Example 5. Synthesis of amino lipid compound 149

[0368] According to the general synthetic method, amino lipid compound 149 (0.73 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.31 g of 149-G as a colorless oil with a purity of 91.68% and a yield of 49%.

[0369] 1H NMR(CDCl3)δ4.87-4.85(m,2H),3.75-3.72(m,2H),3.63-3.61(m,1H),3.45-3.41(m,2 H),3.26-3.22(m,2H),2.61-2.58(m,2H),2.44-2.42(m,2H),2.28-2.25(m,10H),1.61- 1.33(m,20H),1.30-1.25(m,52H),0.92-0.86(m,15H).

[0370] LC-MS (ESI): (M+H) 908.3.

[0371] Example 6. Synthesis of amino lipid compound 154

[0372] According to the general synthetic method, amino lipid compound 154 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.34 g 154-G as a colorless oil with a purity of 93.80% and a yield of 66%.

[0373] 1 H NMR(CDCl3)δ4.87-4.85(m,2H),3.65-3.63(m,1H),3.45-3.43(m,2H),3.40-3.38(m,2H),3.24-3.21(m,2H),2. 44-2.36(m,4H),2.29-2.25(m,10H),1.93-1.89(m,2H),1.60-1.33(m,20H),1.25(m,52H),0.92-0.85(m,15H).

[0374] LC-MS (ESI): (M+H) 922.3.

[0375] Example 7. Synthesis of amino lipid compound 166

[0376] According to the general synthetic method, amino lipid compound 166 (0.33 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.28 g of 144-G as a colorless oil with a purity of 91.31% and a yield of 22%.

[0377] 1H NMR(CDCl3)δ4.87-4.84(m,2H),4.10(d,J=20Hz,1H),3.50-3.47(m,3H),3.09-3.07(m,2H),2.30 -2.25(m,6H),2.22(d,J=3.7Hz,6H),1.61-1.36(m,24H),1.29-1.25(m,52H),0.92-0.85(m,15H).

[0378] LC-MS (ESI): (M+H) 922.3.

[0379] Example 8. Synthesis of amino lipid compound 172

[0380] According to the general synthetic method, amino lipid compound 172 (0.43 g) was prepared as a colorless oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.37 g of 125-G with a purity of 97.49% and a yield of 28%.

[0381] 1 H NMR(CDCl3)δ4.87-4.85(m,2H),3.74-3.72(m,2H),3.64-3.59(m,1H),3.44-3.42(m,2H),3.10-3.06(m, 2H),2.61-2.60(m,2H),2.28-2.23(m,12H),1.62-1.43(m,22H),1.31-1.26(m,52H),0.90-0.86(m,15H).

[0382] LC-MS (ESI): (M+H) 950.4.

[0383] Example 9. Synthesis of amino lipid compound 176

[0384] According to the general synthetic method, amino lipid compound 176 (0.26 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.37 g of 154-G as a colorless oil with a purity of 92.77% and a yield of 17%.

[0385] 1H NMR(CDCl3)δ4.87-4.85(m,2H),3.64-3.62(m,1H),3.45-3.42(m,2H),3.40-3.38(m,2H),3.07-3.05(m,2H),2.41-2.36 (m,2H),2.28-2.25(m,6H),2.22(s,6H),1.91-1.90(m,2H),1.61-1.33(m,24H),1.27-1.25(m,52H),0.92-0.86(m,15H).

[0386] LC-MS (ESI): (M+H) 950.4.

[0387] Example 10. Synthesis of amino lipid compound 119

[0388] According to the general synthetic method, amino lipid compound 119 (0.55 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.28 g of 144-G as a colorless oil with a purity of 89.83% and a yield of 37%.

[0389] 1 H NMR(CDCl3)δ4.86-4.85(m,2H),4.13-4.09(m,2H),3.49-3.48(m,3H),3.13(m,2H) ,2.26-2.21(m,12H),1.74-1.45(m,20H),1.37-1.25(m,54H),0.91-0.85(m,15H).

[0390] LC-MS (ESI): (M+H) 908.4.

[0391] Example 11. Synthesis of amino lipid compound 121

[0392] According to the general synthetic method, amino lipid compound 121 (0.88 g) was prepared as a colorless oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.34 g of 121-G with a purity of 92.94% and a yield of 57%.

[0393] 1H NMR(CDCl3)δ4.86-4.85(m,2H),4.13-4.09(m,2H),3.55-3.46(m,3H),3.14-3.13(m, 2H),2.31-2.21(m,12H),1.75-1.45(m,20H),1.33-1.25(m,58H),0.87-0.85(m,15H).

[0394] LC-MS (ESI): (M+H) 936.5.

[0395] Example 12. Synthesis of amino lipid compound 124

[0396] According to the general synthetic method, amino lipid compound 124 (1.2 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.28 g of 124-G as a colorless oil with a purity of 95.32% and a yield of 79%.

[0397] 1 H NMR(CDCl3)δ4.86-4.85(m,2H),3.75-3.72(m,2H),3.64-3.62(m,1H),3.44-3.42(m,2H),3.15-3.10(m, 2H),2.61-2.59(m,2H),2.28-2.21(m,12H),1.72-1.44(m,20H),1.35-1.25(m,54H),0.91-0.86(m,15H).

[0398] LC-MS (ESI): (M+H) 922.3.

[0399] Example 13. Synthesis of amino lipid compound 129

[0400] According to the general synthetic method, amino lipid compound 129 (0.79 g) was prepared as a colorless oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.34 g of 154-G with a purity of 92.29% and a yield of 51%.

[0401] 1H NMR(CDCl3)δ4.87-4.85(m,2H),3.65(p,J=7.0Hz,1H),3.44(q,J=6.2Hz,2H),3.39(t,J=6.6Hz,2H),3.13-3.1 0(m,2H),2.41-2.22(m,14H),1.97-1.88(m,2H),1.61-1.44(m,20H),1.36-1.25(m,54H),0.92-0.86(m,15H).

[0402] LC-MS (ESI): (M+H) 936.4.

[0403] Example 14. Synthesis of amino lipid compound 131

[0404] According to the general synthetic method, amino lipid compound 131 (1.05 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.40 g of 131-G as a colorless oil with a purity of 94.66% and a yield of 66%.

[0405] 1 H NMR(CDCl3)δ4.88-4.84(m,2H),3.65(p,J=7.0Hz,1H),3.44(q,J=6.2Hz,2H),3.38(t,J=6.7Hz,2H),3.13-3.10(m,2H),2.43-2.3 7(m,2H),2.31-2.22(m,12H),1.95-1.88(m,2H),1.74-1.67(m,2H),1.61-1.44(m,18H),1.32-1.26(m,58H),0.89-0.86(m,15H).

[0406] LC-MS (ESI): (M+H) 964.4.

[0407] Example 15. Synthesis of amino lipid compound 132

[0408] According to the general synthetic method, amino lipid compound 132 (1.08 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.43 g of 132-G as a colorless oil with a purity of 97.18% and a yield of 67%.

[0409] 1H NMR(CDCl3)δ4.88-4.84(m,2H),3.65(p,J=7.0Hz,1H),3.44(q,J=6.2Hz,2H),3.38(t,J=6.6Hz,2H),3.13-3.10(m,2H),2.43-2.3 7(m,2H),2.30-2.22(m,12H),1.95-1.88(m,2H),1.73-1.67(m,2H),1.61-1.44(m,18H),1.30-1.26(m,60H),0.88-0.86(m,15H).

[0410] LC-MS (ESI): (M+H) 978.4.

[0411] Example 16. Synthesis of amino lipid compound 133

[0412] According to the general synthetic method, the amino lipid compound 133 (1.26 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.46 g of 133-G as a colorless oil with a purity of 95.54% and a yield of 77%.

[0413] 1 H NMR(CDCl3)δ4.87-4.83(m,2H),3.64(p,J=7.0Hz,1H),3.43(q,J=6.2Hz,2H),3.37(t,J=6.7Hz,2H),3.12-3.09(m,2H),2.42-2.3 6(m,2H),2.30-2.20(m,12H),1.94-1.87(m,2H),1.72-1.66(m,2H),1.62-1.42(m,18H),1.29-1.24(m,62H),0.87-0.85(m,15H).

[0414] LC-MS (ESI): (M+H) 992.4.

[0415] Example 17. Synthesis of amino lipid compound 036

[0416] According to the general synthesis method, amino lipid compound 036 (1.1 g) was prepared from compound 1522F-4 (3.0 g, 4.07 mmol) and 0.93 g of 130-G as a pale yellow oil with a purity of 96.18% and a yield of 30.3%.

[0417] 1H NMR (600MHz, CDCl3) δ4.95-4.85(m,2H),3.73-3.63(m,1H),3.48(t,J=6.2H z,2H),3.41(t,J=6.7Hz,2H),3.17-3.11(m,2H),2.45(t,J=7.3Hz,1H),2.4 1(t,J=7.3Hz,1H),2.34-2.28(m,6H),2.24(m,6H),2.04-1.86(m,4H),1.79 -1.70(m,2H),1.65-1.43(m,16H),1.38-1.10(m,48H),0.96-0.84(m,15H).

[0418] LC-MS(ESI):(M+H)893.9.

[0419] Example 18. Synthesis of amino lipid compound 037

[0420] According to the general synthesis method, amino lipid compound 037 (0.25 g) was prepared from compound 1522F-5 (0.60 g, 0.75 mmol) and 0.27 g of 130-G as a pale yellow oil with a purity of 95.17% and a yield of 35.4%.

[0421] 1 H NMR (600MHz, CDCl3) δ4.08 (m, 4H), 3.73-3.64 (m, 1H), 3.48 (t, J = 6.1Hz, 2H), 3.42(t,J=6.7Hz,2H),3.15(m,2H),2.45(t,J=7.3Hz,1H),2.42(t,J=7.3Hz,1 H),2.36-2.28(m,4H),2.25(m,6H),1.98-1.87(m,4H),1.78-1.71(m,2H),1. 66-1.55(m,10H),1.52-1.42(m,6H),1.39-1.20(m,56H),0.94-0.84(m,15H).

[0422] LC-MS (ESI): (M+H) 950.0.

[0423] Example 19. Synthesis of amino lipid compound 038

[0424] According to the general method, amino lipid compound 038 (0.65 g) was prepared from compound 1522F-6 (0.82 g, 1.03 mmol) and 0.20 g of 130-G as a light yellow oil with a purity of 95.32% and a yield of 66.2%.

[0425] 1 H NMR(600MHz, CDCl3) δ3.90-3.85(t,J=6.5Hz,4H),3.70-3.62(m,1H),3.47-3.44(m,2 H),3.42-3.40(t,J=6.7Hz,2H),3.13-3.10(m,2H),2.46-2.42(t,J=7.3Hz,1H),2.42- 2.39(t,J=7.3Hz,1H),2.33-2.27(m,6H),2.23(s,6H),1.90-1.86(m,4H),1.67-1.60( m,6H),1.58-1.54(m,2H),1.44-1.40(m,4H),1.36-1.13(m,60H),0.93-0.80(m,15H).

[0426] LC-MS (ESI): (M+H) 950.6.

[0427] Example 20. Synthesis of amino lipid compound 039

[0428] According to the general method, amino lipid compound 039 (0.92 g) was prepared from compound 1522F-7 (2.2 g, 2.59 mmol) and 0.59 g 130-G as a pale yellow oil with a purity of 95.23% and a yield of 35.3%.

[0429] 1 H NMR (600MHz, CDCl3) δ4.91-4.87(m,2H),3.74-3.71(m,1H),3.45-3.42(m,2H),3.41- 3.39(t,J=6.7Hz,2H),3.14-3.03(m,2H),2.54-2.50(m,4H),2.46-2.41(m,4H),2.30 -2.26(m,4H),2.01-1.86(m,4H),1.78-1.69(m,2H),1.67-1.61(m,4H),1.60-1.55(m ,2H),1.52-1.50(m,6H),1.47-1.40(m,4H),1.34-1.20(m,60H),0.92-0.87(m,21H).

[0430] LC-MS (ESI): (M+H) 1006.4.

[0431] Example 21. Synthesis of amino lipid compound 040

[0432] According to the general method, amino lipid compound 040 (2.4 g) was prepared from compound 1522F-8 (4.3 g, 5.23 mmol) and 1.19 g of 130-G as a pale yellow oil with a purity of 95.77% and a yield of 46.9%.

[0433] 1 H NMR (600MHz, CDCl3) δ4.91-4.87(m,2H),3.78-3.76(m,1H),3.48-3.46(m,2H),3.42-3.40(t ,J=6.7Hz,2H),3.17-3.05(m,2H),2.57-2.52(m,4H),2.48-2.40(m,4H),2.32-2.27(m,4H), 2.00-1.86(m,4H),1.77-1.68(m,2H),1.66-1.60(m,4H),1.60-1.56(m,2H),1.53-1.52(m,6 H),1.49-1.41(m,4H),1.35-1.21(m,56H),1.06-1.03(t,J=7.1Hz,6H),0.91-0.88(m,15H).

[0434] LC-MS(ESI):(M+H)978.3.

[0435] Example 22. Synthesis of amino lipid compound 041

[0436] According to the general method, amino lipid compound 041 (0.56 g) was prepared from compound 1522F-9 (1.03 g, 1.4 mmol) and 0.32 g of 130-G as a light yellow oil with a purity of 96.26% and a yield of 44.9%.

[0437] 1H NMR (600MHz, CDCl3) δ4.93-4.83(m,2H),3.78-3.58(m,1H),3.47(t,J=5.9Hz,2H),3.41(t,J=6.7Hz,2H),3.14(m,2H),2.49-2.3 7(m,3H),2.36-2.24(m,11H),1.99-1.87(m,2H),1.80-1.69(m,2H),1.66-1.43(m,18H),1.37-1.22(m,48H),0.97-0.85(m,15H).

[0438] LCMS (ESI): (M+H) 893.9.

[0439] Example 23. Synthesis of amino lipid compound 042

[0440] According to the general method, amino lipid compound 042 (0.93 g) was prepared as a pale yellow oil from compound 1522F-10 (1.91 g, 2.25 mmol) and 0.47 g of 130-G with a purity of 95.58% and a yield of 41.1%.

[0441] 1 H NMR (600MHz, CDCl3) δ4.95-4.82(m,2H),3.73-3.62(m,1H),3.50-3.44(m,2H),3.4 4-3.39(m,2H),3.16-3.13(m,2H),2.44-2.41(m,4H),2.32-2.29(m,4H),2.29(s,4 H),2.26(s,2H),1.98-1.86(m,4H),1.79-1.71(m,2H),1.66-1.61(m,4H),1.60-1. 56(m,2H),1.55-1.51(m,6H),1.47(m,4H),1.38-1.14(m,64H),0.94-0.89(m,15H).

[0442] LCMS (ESI): (M+H) 1006.3.

[0443] Example 24. Synthesis of amino lipid compound 044

[0444] According to the general method, amino lipid compound 044 (0.32 g) was prepared as a pale yellow oil from compound 1522F-11 (1.35 g, 1.49 mmol) and 0.34 g of 130-G with a purity of 95.13% and a yield of 20.2%.

[0445] 1 H NMR (600MHz, CDCl3) δ4.95-4.81(m,2H),3.70-3.66(m,1H),3.49-3.46(m,2H),3.43-3.40 (t,J=6.7Hz,2H),3.23-3.07(m,2H),2.46-2.44(t,J=7.3Hz,1H),2.43-2.40(t,J=7.3Hz, 1H),2.35-2.28(m,6H),2.25(s,6H),2.02-1.93(m,4H),1.80-1.70(m,2H),1.67-1.61(m, 4H),1.61-1.57(m,2H),1.53(m,6H),1.50-1.42(m,4H),1.40-1.15(m,72H),0.91(m,15H).

[0446] LC-MS (ESI): (M+H) 1062.5.

[0447] Example 25. Synthesis of amino lipid compound 045

[0448] According to the general method, amino lipid compound 045 (1.6 g) was prepared from compound 1522F-12 (2.8 g, 3.8 mmol) and 0.86 g of 130-G as a light yellow oil with a purity of 97.38% and a yield of 47.2%.

[0449] 1 H NMR (600MHz, CDCl3) δ4.97-4.84(m,2H),3.70-3.66(m,1H),3.49-3.47(t,J=6.1Hz,2H),3.43- 3.40(t,J=6.7Hz,2H),3.16-3.13(m,2H),2.46-2.44(t,J=7.2Hz,1H),2.43-2.40(t,J=7.3Hz,1 H),2.34-2.27(m,6H),2.25(s,6H),1.98-1.92(m,4H),1.78-1.70(m,2H),1.67-1.61(m,4H),1. 60-1.57(m,2H),1.56-1.53(m,6H),1.48-1.47(m,4H),1.39-1.17(m,48H),0.99-0.86(m,15H).

[0450] LC-MS(ESI):(M+H)893.9.

[0451] Example 26. Synthesis of amino lipid compound 046

[0452] According to the general method, amino lipid compound 046 (0.71 g) was prepared from compound 1522F-13 (1.00 g, 1.31 mmol) and 0.30 g of 130-G as a pale yellow oil with a purity of 95.61% and a yield of 59.0%.

[0453] 1 H NMR(600MHz, CDCl3)δ4.00(d,J=1.8Hz,2H),3.99(d,J=1.8Hz,2H),3.72-3.63(m,1H), 3.49-3.46(m,2H),3.43-3.41(t,J=6.7Hz,2H),3.15(m,2H),2.47-2.44(t,J=7.3Hz,1H ),2.43-2.40(t,J=7.3Hz,1H),2.34-2.29(m,6H),2.25(s,6H),1.91-1.88(m,4H),1.69 -1.61(m,6H),1.61-1.56(m,2H),1.46(m,4H),1.38-1.16(m,56H),0.96-0.84(m,15H).

[0454] LC-MS(ESI):(M+H)Found 922.2.

[0455] Example 27. Synthesis of amino lipid compound 047

[0456] According to the general method, amino lipid compound 047 (0.13 g) was prepared from compound 1522F-14 (0.80 g, 1.09 mmol) and 0.25 g of 130-G as a pale yellow oil with a purity of 96.01% and a yield of 13.4%.

[0457] 1H NMR (600MHz, CDCl3) δ4.94-4.85(m,1H),4.09-4.07(m,2H),3.72-3.63(m,1H),3.49-3.47(t,J=6.0 Hz,2H),3.42-3.40(t,J=6.7Hz,2H),3.16-3.13(m,2H),2.46-2.44(t,J=7.3Hz,1H),2.43-2.40(t,J =7.3Hz,1H),2.35-2.28(m,6H),2.25(s,6H),1.97-1.90(m,2H),1.76-1.71(m,2H),1.67-1.61(m,6H ),1.61-1.56(m,2H),1.55-1.53(m,4H),1.48-1.47(m,4H),1.41-1.20(m,52H),0.95-0.86(m,12H).

[0458] LC-MS(ESI):(M+H)893.9.

[0459] Example 28. Synthesis of amino lipid compound 048

[0460] According to the general method, amino lipid compound 048 (0.276 g) was prepared from compound 1522F-15 (0.56 g, 0.76 mmol) and 0.17 g of 130-G as a pale yellow oil with a purity of 95.01% and a yield of 40.7%.

[0461] 1 H NMR (600MHz, CDCl3) δ4.07 (t, J = 6.8 Hz, 4H), 3.71-3.62 (m, 1H), 3.48-3.46 (t, J =6.2Hz,2H),3.42-3.40(t,J=6.7Hz,2H),3.16-3.12(m,2H),2.46-2.43(t,J=7.2Hz,1H),2.42-2.40(t,J=7.3Hz,1H),2.33-2.28(m,6 H),2.24(s,6H),2.01-1.89(m,2H),1.77-1.69(m,2H),1.67-1.55(m,10H),1.47-1.46(m,4H),1.40-1.21(m,56H),0.93-0.89(m,9H).

[0462] LC-MS(ESI):(M+H)893.9.

[0463] Example 29. Synthesis of amino lipid compound 049

[0464] According to the general method, amino lipid compound 049 (0.058 g) was prepared from compound 1522F-16 (0.16 g, 0.21 mmol) and 0.086 g of 130-G as a pale yellow oil with a purity of 95.29% and a yield of 30.1%.

[0465] 1 H NMR (600MHz, CDCl3) δ4.09 (m, 4H), 3.73-3.64 (m, 1H), 3.48 (t, J = 6.1Hz, 2H), 3.42(t,J=6.7Hz,2H),3.15(m,2H),2.46(t,J=7.3Hz,1H),2.42(t,J=7.3Hz,1 H),2.38-2.28(m,4H),2.26(m,6H),2.05-1.89(m,4H),1.78-1.71(m,2H),1. 68-1.56(m,10H),1.53-1.43(m,6H),1.39-1.18(m,52H),0.96-0.85(m,15H).

[0466] LC-MS (ESI): (M+H) 922.0.

[0467] Example 30. Synthesis of amino lipid compound 122

[0468] According to the general synthetic method, the amino lipid compound 122 (0.51 g) was prepared from compound 1522F-1 (1 g, 1.26 mmol) and 0.33 g 122-G as a pale yellow oil with a purity of 92.94% and a yield of 43%.

[0469] 1 H NMR in CDCl3δ4.86-4.85(m,2H),4.13-4.09(m,2H),3.55(m,1H),3.50-3.46(m,2H),3.13 (m,2H),2.30-2.21(m,12H),1.74-1.45(m,20H),1.25(m,60H),0.88-0.85(m,15H).

[0470] LC-MS(ESI):(M+H)950.5.

[0471] Example 31. Synthesis of amino lipid compound 123

[0472] According to the general synthetic method, the amino lipid compound 123 (0.88 g) was prepared as a pale yellow oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.46 g of 123-G with a purity of 90.77% and a yield of 55%.

[0473] 1 H NMR in CDC13δ4.87-4.85(m,2H),4.13-4.09(m,2H),3.56-3.53(m,1H),3.50-3.46(m,2H),3.14-3. 11(m,2H),2.38-2.22(m,12H),1.77-1.44(m,20H),1.27-1.25(m,62H),0.88-0.85(m,15H).

[0474] LC-MS (ESI): (M+H) 964.5.

[0475] Example 32. Synthesis of amino lipid compound 126

[0476] According to the general synthetic method, the amino lipid compound 126 (0.83 g) was prepared as a pale yellow oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.43 g of 126-G with a purity of 93.73% and a yield of 53%.

[0477] 1H NMR in CDCl3δ4.89-4.84(m,2H),3.76-3.72(m,2H),3.63(m,1H),3.45-3.41(m,2H),3.15-3.10 (m,2H),2.62-2.59(m,2H),2.31-2.22(m,12H),1.72-1.26(m,78H),0.89-0.86(m,15H).

[0478] LC-MS (ESI): (M+H) 950.3.

[0479] Example 33. Synthesis of amino lipid compound 127

[0480] According to the general synthetic method, the amino lipid compound 127 (0.75 g) was prepared as a pale yellow oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.46 g of 127-G with a purity of 95.58% and a yield of 48%.

[0481] 1H NMR in CDCl3δ4.88-4.84(m,2H),3.75-3.72(m,2H),3.62(m,1H),3.44-3.40(m,2H),3.14-3.09(m,2H ),2.61-2.59(m,2H),2.30-2.24(m,6H),2.21(s,6H),1.70-1.25(m,80H),0.88-0.86(m,15H).

[0482] LC-MS (ESI): (M+H) 964.4.

[0483] Example 34. Synthesis of amino lipid compound 128

[0484] According to the general synthetic method, the amino lipid compound 128 (0.76 mg) was prepared as a pale yellow oil from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.49 g of 128-G with a purity of 94.70% and a yield of 47%.

[0485] 1 H NMR in CDCl3δ4.88-4.84(m,2H),3.75-3.72(m,2H),3.62(m,1H),3.44-3.40(m,2H),3.15-3.09(m,2H),2.61-2 .59(m,2H),2.29-2.25(m,6H),2.21(s,6H),1.71-1.43(m,20H),1.27-1.25(m,62H),0.88-0.86(m,15H).

[0486] LC-MS (ESI): (M+H) 978.4.

[0487] Example 35. Synthesis of amino lipid compound 145

[0488] According to the general synthetic method, the amino lipid compound 145 (0.24 g) was prepared as a pale yellow oil from compound 1522F-2 (1 g, 1.28 mmol) and 0.28 g of 120-G with a purity of 90.77% and a yield of 20%.

[0489] 1H NMR (600MHz, CDCl3) δ4.89-4.82(m,2H),4.12(d,J=34.5Hz,2H),3.59-3.44(m,3H),3.32-3.20(m,2H),2.43(dd,J=36.8,28.9Hz,2H),2.32 (s,4H),2.26(td,J=7.6,3.5Hz,6H),1.60(dd,J=11.6,6.9Hz,6H),1.54-1.40(m,12H),1.36-1.12(m,56H),0.88(dt,J=17.2,7.0Hz,15H).

[0490] LC-MS (ESI): (M+H) 908.3

[0491] Example 36. Synthesis of amino lipid compound 146

[0492] According to the general synthetic method, the amino lipid compound 146 (0.55 g) was prepared as a pale yellow oil from compound 1522F-2 (1 g, 1.28 mmol) and 0.31 g of 121-G with a purity of 97.28% and a yield of 47%.

[0493] 1 H NMR(600MHz, CDCl3) δ4.86(p,J=6.5Hz,2H),4.12(d,J=34.8Hz,2H),3.59-3.45(m,3H),3.25(dd,J=14.6,5.8Hz,2H),2 .52-2.37(m,2H),2.33-2.22(m,10H),1.65-1.55(m,6H),1.55-1.39(m,12H),1.38-1.14(m,58H),0.91-0.83(m,15H).

[0494] LC-MS (ESI): (M+H) 922.3.

[0495] Example 37. Synthesis of amino lipid compound 147

[0496] According to the general synthetic method, the amino lipid compound 147 (0.92 g) was prepared as a pale yellow oil from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g of 122-G with a purity of 92.84% and a yield of 59%.

[0497] 1H NMR (600MHz, CDCl3) δ4.89-4.81(m,2H),4.12(d,J=34.8Hz,2H),3.59-3.42(m,3H),3.24(dd,J=8.7,6.6Hz,2H),2.51-2.3 8(m,2H),2.33-2.22(m,10H),1.64-1.54(m,6H),1.55-1.40(m,12H),1.26(dd,J=8.2,4.6Hz,60H),0.87(t,J=7.0Hz,15H).

[0498] LC-MS (ESI): (M+H) 936.3.

[0499] Example 38. Synthesis of amino lipid compound 148

[0500] According to the general synthetic method, the amino lipid compound 148 (0.85 g) was prepared as a pale yellow oil from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.47 g of 123-G with a purity of 96.59% and a yield of 54%.

[0501] 1 H NMR (600MHz, CDCl3) δ4.86 (p, J=6.6Hz, 2H), 4.12 (d, J=34.9Hz, 2H), 3.59-3.43 (m, 3H), 3.24 (dd, J=8.8, 6.6Hz, 2H), 2.49-2. 37(m,2H),2.32-2.22(m,10H),1.65-1.54(m,6H),1.54-1.41(m,12H),1.27(dd,J=15.6,9.8Hz,62H),0.87(t,J=7.0Hz,15H).

[0502] LC-MS (ESI): (M+H) 950.3.

[0503] Example 39. Synthesis of amino lipid compound 150

[0504] According to the general synthetic method, amino lipid compound 150 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.40 g of 125-G as a pale yellow oil with a purity of 96.39% and a yield of 65%.

[0505] 1H NMR (600MHz, CDCl3) δ4.89-4.82(m,2H),3.77-3.69(m,2H),3.66-3.56(m,1H),3.46-3.36(m,2H),3.29-3.18(m,2H),2.63-2.56(m,2H) ,2.49-2.39(m,2H),2.32-2.21(m,10H),1.52(dd,J=40.8,21.6,14.0,6.9Hz,18H),1.34-1.13(m,56H),0.87(dt,J=14.0,7.0Hz,15H).

[0506] LC-MS (ESI): (M+H) 922.3.

[0507] Example 40. Synthesis of amino lipid compound 151

[0508] According to the general synthetic method, the amino lipid compound 151 (1.0 g) was prepared as a pale yellow oil from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g of 126-G with a purity of 95.27% and a yield of 64%.

[0509] 1 H NMR (600MHz, CDCl3) δ4.89-4.82(m,2H),3.77-3.70(m,2H),3.66-3.57(m,1H),3.46-3.38(m,2H),3.25(dd,J=10.9,4.8Hz,2H),2.64- 2.56(m,2H),2.44(dd,J=16.0,8.7Hz,2H),2.34-2.22(m,10H),1.65-1.37(m,18H),1.27(dd,J=12.0,6.0Hz,58H),0.91-0.84(m,15H).

[0510] LC-MS (ESI): (M+H) 936.3.

[0511] Example 41. Synthesis of amino lipid compound 152

[0512] According to the general synthetic method, amino lipid compound 152 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.47 g 127-G as a pale yellow oil with a purity of 95.65% and a yield of 63%.

[0513] 1H NMR (600MHz, CDCl3) δ4.89-4.82(m,2H),3.78-3.70(m,2H),3.67-3.56(m,1H),3.48-3.36(m,2H),3.30-3.19(m,2H),2.64-2. 56(m,2H),2.43(dd,J=15.6,7.5Hz,2H),2.31-2.22(m,10H),1.66-1.36(m,18H),1.35-1.11(m,60H),0.87(t,J=7.0Hz,15H).

[0514] LC-MS (ESI): (M+H) 950.4.

[0515] Example 42. Synthesis of amino lipid compound 153

[0516] According to the general synthetic method, the amino lipid compound 153 (1.07 g) was prepared as a pale yellow oil from compound 1522F-2 (1.5 g, 1.92 mmol) and 0.58 g of 128-G with a purity of 93.89% and a yield of 58%.

[0517] 1 H NMR (600MHz, CDCl3) δ4.90-4.82(m,2H),3.77-3.70(m,2H),3.65-3.57(m,1H),3.47-3.38(m,2H),3.25(dd,J=9.5,5.8Hz,2H),2.59(dd,J=9.2, 5.5Hz,2H),2.51-2.40(m,2H),2.31(s,4H),2.26(dt,J=11.7,3.6Hz,6H ),1.64-1.36(m,18H),1.26(d,J=12.4Hz,62H),0.87(t,J=6.8Hz,15H).

[0518] LC-MS (ESI): (M+H) 964.4.

[0519] Example 43. Synthesis of amino lipid compound 155

[0520] According to the general synthetic method, amino lipid compound 155 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g 130-G as a pale yellow oil with a purity of 93.39% and a yield of 64%.

[0521] 1H NMR (600MHz, CDCl3) δ4.89-4.82(m,2H),3.64(p,J=6.6Hz,1H),3.43(q,J=6.0Hz,2H),3.38(t,J=6.7Hz,2H),3.28-3.18(m,2H),2.52-2.3 4(m,4H),2.34-2.21(m,10H),1.96-1.84(m,2H),1.52(ddt,J=21.3,13.8,6.6Hz,18H),1.35-1.07(m,56H),0.87(dt,J=7.2,6.2Hz,15H).

[0522] LC-MS (ESI): (M+H) 936.4.

[0523] Example 44. Synthesis of amino lipid compound 156

[0524] According to the general synthetic method, the amino lipid compound 156 (1.0 g) was prepared from compound 1522F-2 (1.5 g, 1.92 mmol) and 0.54 g of 131-G as a pale yellow oil with a purity of 96.96% and a yield of 44%.

[0525] 1 H NMR (600MHz, CDCl3) δ4.86(p,J=5.7Hz,2H),3.64(p,J=6.8Hz,1H),3.43(q,J=6.1Hz,2H),3.38(t,J=6.7Hz,2H),3.22(dd,J=15.6,1 1.5Hz,2H),2.48-2.34(m,4H),2.32-2.21(m,10H),1.96-1.85(m,2H),1.66-1.37(m,18H),1.34-1.18(m,58H),0.91-0.82(m,15H).

[0526] LC-MS (ESI): (M+H) 950.3.

[0527] Example 45. Synthesis of amino lipid compound 157

[0528] According to the general synthetic method, the amino lipid compound 157 (1.1 g) was prepared from compound 1522F-2 (1.5 g, 1.92 mmol) and 0.58 g of 132-G as a pale yellow oil with a purity of 90.77% and a yield of 59%.

[0529] 1H NMR (600MHz, CDCl3) δ4.89-4.81(m,2H),3.64(p,J=6.6Hz,1H),3.43(qd,J=6.0,1.1Hz,2H),3.38(t,J=6.7Hz,2H),3.27- 3.18(m,2H),2.49-2.34(m,4H),2.32-2.22(m,10H),1.96-1.84(m,2H),1.51(m,18H),1.26(m,60H),0.91-0.81(m,15H).

[0530] LC-MS (ESI): (M+H) 964.4.

[0531] Example 46. Synthesis of amino lipid compound 158

[0532] According to the general synthetic method, the amino lipid compound 158 (0.96 g) was prepared as a pale yellow oil from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.54 g of 133-G with a purity of 97.59% and a yield of 59%.

[0533] 1 H NMR (600MHz, CDCl3) δ4.85(m,2H),3.68-3.59(m,1H),3.47-3.40(m,2H),3.37(m,2H),3.28-3.19(m,2H),2.50-2 .34(m,4H),2.30(s,4H),2.28-2.21(m,6H),1.90(m,2H),1.65-1.37(m,18H),1.21(m,62H),0.92-0.80(m,15H).

[0534] LC-MS (ESI): (M+H) 978.4.

[0535] Example 47. Synthesis of amino lipid compound 167

[0536] According to the general synthetic method, the amino lipid compound 167 (0.45 g) was prepared as a pale yellow oil from compound 1522F-3 (1 g, 1.24 mmol) and 0.27 g of 120-G with a purity of 92.68% and a yield of 39%.

[0537] 1H NMR (600MHz, CDCl3) δ4.92-4.79(m,2H),4.13(s,0.6H),4.10(s,1.4H),3.61-3.42(m,3H),3.13-3.04(t,J=6.0H z,2H),2.35-2.26(m,6H),2.25(s,4H),2.23(s,2H),1.68-1.31(m,26H),1.31-1.16(m,52H),0.93-0.82(m,15H).

[0538] LC-MS (ESI): (M+H) 936.2.

[0539] Example 48. Synthesis of amino lipid compound 168

[0540] According to the general synthetic method, the amino lipid compound 168 (0.65 g) was prepared as a pale yellow oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.39 g of 121-G with a purity of 91.4% and a yield of 42%.

[0541] 1 H NMR (600MHz, CDCl3) δ4.97-4.79(m,2H),4.16(s,0.6H),4.14(s,1.4H),3.62-3.44(m,3H),3.11(t,J=6 .0Hz,2H),2.35-2.28(m,6H),2.25(s,6H),1.68-1.35(m,24H),1.42-1.16(m,56H),0.93-0.87(m,15H).

[0542] LC-MS(ESI):(M+H)950.1.

[0543] Example 49. Synthesis of amino lipid compound 169

[0544] According to the general synthetic method, the amino lipid compound 169 (0.58 g) was prepared as a pale yellow oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 122-G with a purity of 91.97% and a yield of 37%.

[0545] 1H NMR (600MHz, CDCl3) δ4.93-4.81(m,2H),4.16(s,0.6H),4.13(s,1.4H),3.63-3.48(m,3H),3.16-3.07(t,J= 6.0Hz,2H),2.35-2.28(m,6H),2.26(s,6H),1.69-1.42(m,22H),1.40-1.17(m,60H),0.91(t,J=6.8Hz,15H).

[0546] LC-MS(ESI):(M+H)964.1.

[0547] Example 50. Synthesis of amino lipid compound 170

[0548] According to the general synthetic method, the amino lipid compound 170 (0.41 g) was prepared as a pale yellow oil from compound 1522F-3 (1 g, 1.24 mmol) and 0.35 g of 123-G with a purity of 92.56% and a yield of 34%.

[0549] 1 H NMR (600MHz, CDCl3) δ4.94-4.84(m,2H),4.16(s,0.6H),4.13(s,1.4H),3.62-3.48(m,3H),3.15-3.08(t,J=6.0Hz, 2H),2.38-2.29(m,6H),2.28(s,4H),2.26(s,2H),1.71-1.42(m,22H),1.41-1.16(m,62H),0.91(t,J=7.0Hz,15H).

[0550] LC-MS (ESI): (M+H) 978.1.

[0551] Example 51. Synthesis of amino lipid compound 171

[0552] According to the general synthetic method, the amino lipid compound 171 (0.52 g) was prepared as a pale yellow oil from compound 1522F-3 (1 g, 1.24 mmol) and 0.27 g of 149-G with a purity of 91.21% and a yield of 46%.

[0553] 1H NMR (600MHz, CDCl3) δ4.92-4.79(m,2H),3.74(q,J=7.1Hz,2H),3.66-3.57(m,1H),3.44(q,J=6.6Hz,2H),3.10-3.06(m, 2H), 2.60 (t, J = 7.0Hz, 2H), 2.30-2.24 (m, 6H), 2.22 (s, 6H), 1.65-1.33 (m, 22H), 1.32-1.17 (m, 54H), 0.95-0.82 (m, 15H).

[0554] LC-MS (ESI): (M+H) 936.1.

[0555] Example 52. Synthesis of amino lipid compound 173

[0556] According to the general synthetic method, the amino lipid compound 173 (0.42 g) was prepared as a pale yellow oil from compound 1522F-3 (1 g, 1.24 mmol) and 0.32 g of 126-G with a purity of 92.16% and a yield of 36%.

[0557] 1 H NMR (600MHz, CDCl3) δ4.91-4.81(m,2H),3.74(q,J=7.1Hz,2H),3.66-3.57(m,1H),3.43(q,J=6.6Hz,2H),3.13-3.01(m, 2H),2.60(t,J=7.0Hz,2H),2.30-2.24(m,6H),2.22(s,6H),1.64-1.37(m,22H),1.35-1.16(m,58H),0.95-0.84(m,15H).

[0558] LC-MS(ESI):(M+H)964.1.

[0559] Example 53. Synthesis of amino lipid compound 174

[0560] According to the general synthetic method, the amino lipid compound 174 (0.53 g) was prepared as a pale yellow oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.45 g of 127-G with a purity of 97.18% and a yield of 34%.

[0561] 1H NMR (600MHz, CDCl3) δ4.92-4.81(m,2H),3.81-3.70(m,2H),3.70-3.61(m,1H),3.51-3.41(m,2H),3.14-3.10(m,2H),2.63(t,J=7.0Hz,2H), 2.58(t,J=7.1Hz,0.5H),2.47-2.41(t,J=6.9Hz,1.5H),2.38-2.28(m,10H),1.71-1.40(m,22H),1.38-1.17(m,60H),0.91(t,J=7.0Hz,15H).

[0562] LC-MS (ESI): (M+H) 978.2.

[0563] Example 54. Synthesis of amino lipid compound 175

[0564] According to the general synthetic method, the amino lipid compound 175 (0.6 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.49 g of 128-G as a pale yellow oil with a purity of 91.10% and a yield of 38%.

[0565] 1 H NMR (600MHz, CDCl3) δ4.89-4.81(m,2H),3.81-3.70(m,2H),3.67-3.55(m,1H),3.47-3.38(m,2H),3.14-3.05(m,2H),2.60(t,J=7.0Hz,2H ),2.54(t,J=7.1Hz,0.5H),2.43(t,J=6.9Hz,1.5H),2.36-2.24(m,10H),1.69-1.36(m,22H),1.35-1.10(m,62H),0.88(t,J=7.0Hz,15H).

[0566] LC-MS(ESI):(M+H)992.2.

[0567] Example 55. Synthesis of amino lipid compound 177

[0568] According to the general synthetic method, the amino lipid compound 177 (0.6 g) was prepared as a pale yellow oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 130-G with a purity of 92.65% and a yield of 39%.

[0569] 1H NMR (600MHz, CDCl3) δ4.94-4.80(m,2H),3.69-3.56(m,1H),3.48-3.41(m,2H),3.41-3.35(m,2H),3.07(t,J=7.0Hz,2H) ,2.46-2.35(m,3H),2.33-2.23(m,11H),1.96-1.87(m,2H),1.68-1.38(m,24H),1.36-1.14(m,54H),0.96-0.85(m,15H).

[0570] LC-MS (ESI): (M+H) 964.2.

[0571] Example 56. Synthesis of amino lipid compound 178

[0572] According to the general synthetic method, the amino lipid compound 178 (0.43 g) was prepared as a pale yellow oil from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 131-G with a purity of 94.43% and a yield of 27%.

[0573] 1 H NMR (600MHz, CDCl3) δ4.90-4.84(m,2H),3.70-3.58(m,1H),3.48-3.43(m,2H),3.43-3.38(m,2H),3.08(t,J=7.0Hz,2H) ,2.46-2.35(m,3H),2.33-2.23(m,11H),1.96-1.85(m,2H),1.66-1.39(m,24H),1.36-1.10(m,56H),0.92-0.83(m,15H).

[0574] LC-MS (ESI): (M+H) 978.2.

[0575] Comparative Example 1. Synthesis of Compound 118

[0576] According to the general synthetic method, compound 118 (0.85 g) was prepared from compound 1522F-1 1.3 g (1.3 g, 1.64 mmol) and 0.31 g 118-G as a colorless oil with a purity of 90.77% and a yield of 57%.

[0577] 1H NMR in(CDCl3)δ4.86-4.85(m,2H),3.62(m,1H),3.11-3.09(m,2H),2.30-2.26( m,8H),2.22(s,6H),1.70-1.44(m,20H),1.25(m,60H),0.88-0.86(m,15H).

[0578] LC-MS (ESI): (M+H) 920.5.

[0579] Biological tests

[0580] The structures of compounds 118 and 1004 used in the biological tests disclosed herein are shown below:

[0581] Compound 118 can be prepared according to the synthesis method of compound 10 (Example 9) described in Comparative Example 1 or Chinese Patent CN107922364A; Compound 1004 can be prepared according to the synthesis method of compound 7 (Example 7) in CN114127044A.

[0582] Experimental Example 1. Preparation of lipid nanoparticles encapsulating luciferase mRNA (Fluc mRNA)

[0583] (1) Ingredients:

[0584] A specified amount of Fluc mRNA stock solution, 0.2 M sodium acetate buffer, and DEPC water were added to a container and mixed to prepare an aqueous phase.

[0585] The amino lipid compound, helper lipid, structural lipid, and PEG-lipid disclosed herein were dissolved in anhydrous ethanol to prepare respective solutions at concentrations of 20 mg / mL, 10 mg / mL, 20 mg / mL, and 25 mg / mL, respectively. The molar ratio of amino lipid compound: DSPC: CHO-HP: PEG2000-DMG was 48:10:40.5:1.5. Each of the four solutions was pipetted and mixed to prepare an alcohol phase.

[0586] (2) Encapsulation: mRNA-encapsulated lipid nanoparticles (mRNA-LNPs) were obtained by injecting the mRNA into the microfluidic chip at a flow rate of 12 mL / min:4 mL / min for aqueous phase:alcohol phase, and encapsulating the mRNA at a flow rate of 9 mL / min:3 mL / min for aqueous phase:alcohol phase.

[0587] (3) Dialysis: The product from step (2) was placed in a dialysis bag and placed in a Tris buffer-8% sucrose solution for displacement to remove residual ethanol, unassembled lipids, and other components. Dialysis was performed at room temperature in the dark under magnetic stirring for 2 hours (dialysis solution was changed every hour).

[0588] (4) The product of step (3) was sterilized by passing it through a 0.22 μm microporous filter membrane and then aliquoted.

[0589] A lipid nanoparticle preparation encapsulating Fluc mRNA was prepared, wherein the concentration of Fluc mRNA was 0.2 μg / μL, the mass ratio of Fluc mRNA:Lipid was 1:10, the particle size was 80-130 nm, and the encapsulation efficiency was above 80%.

[0590] Experimental Example 2. Evaluation of in vivo delivery performance of lipid nanoparticles

[0591] Animal Preparation: Female BALB / c mice, 6-8 weeks old, were housed in an SPF-grade enclosure. Animal experiments were conducted in strict accordance with national health agency guidelines and animal ethics requirements.

[0592] In vivo delivery: Before injection of the test LNP formulation, gently invert the LNP formulation repeatedly to thoroughly mix the formulation sample. Aspirate the corresponding amount of formulation sample using a 1 ml insulin syringe and inject the LNP formulation via tail vein (IV) injection. Repeat for each formulation in three mice, injecting each mouse with 75 μL of the lipid nanoparticle formulation encapsulating luciferase mRNA (Fluc mRNA) prepared in Experimental Example 1.

[0593] Six hours after LNP injection, mice were injected with 200 μL of D-Luciferin luciferase substrate (Cat. No. 122799; Manufacturer: Perkin Elmer). After substrate injection, the mice were anesthetized by injecting 200 μL of 2.5% avertin, and the injection time of the luciferase substrate was recorded. Ten minutes after substrate injection, the animals were placed in a supine position, and the signal distribution and expression intensity of luciferase in the living animals and various organs were observed under the IVIS in vivo imaging system.

[0594] The fluorescence expression intensity induced by lipid nanoparticles containing luciferase mRNA (Fluc mRNA) encapsulated by representative amino lipid compounds is shown in Table 4, wherein amino lipid compound 118 was used as a control.

[0595] Table 4

[0596] Table 5 shows the ratio of the fluorescence expression intensity induced by lipid nanoparticles of luciferase mRNA (Fluc mRNA) encapsulated with representative amino lipid compounds to the spleen delivery / total delivery in vivo (spleen / total flux) relative to compound 118.

[0597] Table 5

[0598] As can be seen from Table 5, compared with compound 118, except for amino lipid compounds 145, 146, 150, 151, 156, and 157, other amino lipid compounds showed stronger spleen delivery preference.

[0599] Experimental Example 3. Evaluation of the delivery efficiency of lipid nanoparticles to immune cell populations in different tissues and organs

[0600] C57BL / 6 mice (3 per group) were intravenously injected with mRNA-LNP encapsulated with eGFP. Six hours later, the mice were dissected and the spleen, axillary lymph nodes, peripheral blood, bone marrow, lungs and liver were removed to prepare single-cell suspensions of each tissue and organ. After blocking with Fc blocking antibodies and surface staining with flow cytometric fluorescent antibodies, flow cytometry was performed to analyze the expression levels of eGFP in each immune cell population, thereby confirming the delivery efficiency of lipid nanoparticles containing different amino lipid compounds to immune cell populations in different tissues and organs.

[0601] The test results are shown in Figures 1A to 1F. It can be seen from the figures that lipid nanoparticles containing amino lipid compounds 130, 131, and 133 have obvious delivery preferences for immune cells in various tissues and organs.

[0602] Various modifications of the present disclosure, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

Claims

1. An amino lipid compound having a structure of formula (I): or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in: Z1, Z2, Z5, Z6, Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS- or a bond; Z3 is -C(=O)- or a bond; Z4 is -O-, -CH(OH)-; A1, A2, A3, A4, A5, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond; R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle; R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; and R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; Preferably, A5, Z5 and Z6 are bonds.

2. The amino lipid compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: A1 and A2 are each independently a C1-C8 alkylene group or a bond, such as a C1-C6 alkylene group or a bond, or a C1-C4 alkylene group or a bond; Preferably, A1 and A2 are each independently C1-C8 alkylene, C2-C8 alkenylene or C2-C8 alkynylene; Preferably, A1 and A2 are each independently C1-C8 alkylene, such as C1-C6 alkylene, C1-C4 alkylene, C2-C4 alkylene or C1-C3 alkylene; Preferably, A1 and A2 are both bonds; and / or A3 is C1-C 12 alkylene group or bond; Preferably, A3 is a linear C1-C 12 alkylene; Preferably, A3 is a linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 Alkynylidene; Preferably, A3 is a linear C1-C5 alkylene group, such as a linear C1, C2, C3, C4 or C5 alkylene group; Preferably, A3 is -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-; and / or A4 is C1-C 12 alkylene group or bond; Preferably, A4 is a linear C1-C 12 alkylene; Preferably, A4 is a linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 Alkynylidene; Preferably, A4 is a linear C1-C7 alkylene group, such as a linear C1, C2, C3, C4, C5, C6 or C7 alkylene group; Preferably, A4 is a linear C1-C3 alkylene group, such as a linear C1, C2 or C3 alkylene group; Preferably, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-; and / or A5 is a C1-C8 alkylene group or a bond, such as a C1-C6 alkylene group or a bond, a C1-C4 alkylene group or a bond, or a C1-C3 alkylene group or a bond; Preferably, A5 is C1-C8 alkylene, C2-C8 alkenylene or C2-C8 alkynylene; Preferably, A5 is C1-C8 alkylene, such as C1-C8 alkylene, C1-C6 alkylene, C1-C4 alkylene, C2-C4 alkylene or C1-C3 alkylene; Preferably, A5 is a bond; and / or A6 and A7 are each independently C1-C 12 alkylene; Preferably, A6 and A7 are each independently a linear C1-C 12 alkylene; Preferably, A6 and A7 are each independently a linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 Alkynylidene; Preferably, A6 and A7 are each independently a linear C1-C 12 Alkylene, such as straight-chain C2-C 12 Alkylene, C4-C 10 Alkylene, C6-C8 alkylene, C6-C 10 Alkylene or C4-C8 alkylene; Preferably, A6 and A7 are each independently a linear C6-C 10 Alkylene, such as linear C6, C7, C8, C9 or C 10 alkylene; Preferably, A6 and A7 are each independently -(CH2)6-, -(CH2)7-, -(CH2)8- or -(CH2)9-; and / or R1 and R2 are each independently: H; C1-C 18 Hydrocarbon groups, such as C1-C 16 Hydrocarbon, C1-C 12 Hydrocarbon, C1-C 10 alkyl, C1-C8 hydrocarbon, C1-C6 hydrocarbon or C1-C4 hydrocarbon; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or a 3- to 7-membered heterocycle, such as a 4- to 6-membered heterocycle; or, R1 and R2, together with the nitrogen atom to which they are attached, form a 4- to 7-membered heterocycle, such as a 5- to 6-membered heterocycle; Preferably, R1 and R2 are each independently: H; C1-C 18 Alkylene, C2-C 18 Alkenylene or C2-C 18 Alkynylidene; Preferably, R1 and R2 are each independently C1-C 18 Alkyl, such as C1-C4 alkyl; C3-C6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; phenyl; or a 5- to 6-membered heterocyclic ring; or R1 and R2, together with the nitrogen atom to which they are attached, form a 5- to 6-membered heterocyclic ring, such as pyrrolidine, piperidine, piperazine or morpholine; Preferably, R1 and R2 are each independently C1-C4 alkyl, such as C1, C2, C3 or C4 alkyl; Preferably, R1 and R2 are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl or tert-butyl; Preferably, R1 and R2 are each independently C1-C3 alkyl, such as C1, C2 or C3 alkyl; Preferably, R1 and R2 are each independently methyl, ethyl, or n-propyl; and / or R3 is: H; C1-C 18 Hydrocarbon groups, such as C1-C 16 Hydrocarbon, C2-C 12 Hydrocarbon, C4-C 10 alkyl, or C4-C8 alkyl; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or 3- to 7-membered heterocycle, such as 4- to 6-membered heterocycle; Preferably, R3 is C1-C 18 Alkyl, C2-C 18 Alkenyl or C2-C 18 Alkynyl; Preferably, R3 is C1-C 18 Alkyl groups, such as C2-C 10 Alkyl, C4-C8 alkyl; C3-C6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; phenyl; or a 5- to 6-membered heterocyclic ring; Preferably, R3 is a linear C4-C8 alkyl group, such as a linear C4, C5, C6, C7 or C8 alkyl group; Preferably, R3 is n-butyl, n-pentyl, n-hexyl, n-heptyl or n-octyl; and / or Z1, Z2, Z5 and Z6 are each independently -C(=O)-, -O(C=O)-, -O-, or a bond; Preferably, Z1, Z2, Z5 and Z6 are each independently a bond; and / or Z3 is -C(=O)-; and / or Z4 is -O-, or Z4 is -CH(OH)-; and / or Z7 and Z8 are each independently -(C=O)O-, -O(C=O)- or a bond.

3. The amino lipid compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein one of Z7 or Z8 is -(C=O)O-, -O(C=O)- or a bond.

4. The amino lipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is -(C=O)O-; Preferably, the (C═O) in Z7 is connected to A6, and the (C═O) in Z8 is connected to A7.

5. The amino lipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is -O(C=O)-; Preferably, (C═O) in Z7 is connected to R4, and (C═O) in Z8 is connected to R5.

6. The amino lipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein one of Z7 or Z8 is a bond; Preferably, wherein: Z7 is a bond, and Z8 is -(C=O)O-; preferably, the (C=O) in Z8 is connected to A7; or Z7 is a bond, and Z8 is -O(C=O)-; preferably, the (C=O) in Z8 is connected to R5; or Z8 is a bond, and Z7 is -(C=O)O-; preferably, the (C=O) in Z7 is connected to A6; or Z8 is a bond, and Z7 is -O(C=O)-; preferably, the (C=O) in Z7 is connected to R4.

7. The amino lipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein each of Z7 and Z8 is a bond.

8. according to the amino lipid compound described in any one of claims 1 to 7, wherein R4 and R5 are each independently C1-C 20 alkyl; C3-C7 cycloalkyl, such as C3-C6 cycloalkyl or C5-C6 cycloalkyl; phenyl; or a 3- to 7-membered heterocycle, such as a 4- to 6-membered heterocycle; Preferably, R4 and R5 are each independently C1-C 20 hydrocarbon group; C3-C6 cycloalkyl group, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; phenyl group; or a 5- to 6-membered heterocyclic ring; Preferably, R4 and R5 are each independently C1-C 20 Hydrocarbon group.

9. The amino lipid compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently C1-C 20 alkyl.

10. The amino lipid compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently a branched C3-C 20 alkyl; Preferably, R4 and R5 are each independently a branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13 alkyl; Preferably, R4 is a branched alkyl group whose branching occurs at the ɑ, β or γ position relative to Z7; R5 is a branched alkyl group whose branching occurs at the ɑ, β or γ position relative to Z8; Preferably, R4 is a branched alkyl group whose branching occurs at the ɑ or β position relative to Z7; R5 is a branched alkyl group whose branching occurs at the ɑ or β position relative to Z8; Preferably, R4 and R5 are each independently one of the following structures:

11. The amino lipid compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures:

12. The amino lipid compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently 13. The amino lipid compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: Z3 is -C(=O)-; and A4 is C1-C 12 alkylene; Preferably, A4 is a linear C1-C 12 Alkylene, such as straight chain C1-C 10 Alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C4 alkylene, or C1-C3 alkylene; Preferably, A4 is a linear C1-C4 alkylene group, such as a linear C1, C2, C3 or C4 alkylene group; Preferably, A4 is -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-; and / or Z4 is -O-; or Z4 is -CH(OH)-.

14. The amino lipid compound according to any one of claims 1 to 13, having a structure of formula (II) or formula (III): or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in: Z7 and Z8 are each independently -C(=O)-, -CH(OH)-, -C=C-, -C≡C-, -O-, -(C=O)O-, -O(C=O)-, -C(=O)S-, -SC(=O)-, -SS-, or a bond; A3, A4, A6 and A7 are each independently C1-C 12 alkylene, cycloalkyl, phenyl, heterocycle or bond; R1 and R2 are each independently H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocycle; R3 is H or C1-C 18 Hydrocarbon, or cycloalkyl, phenyl, heterocycle; and R4 and R5 are each independently C1-C 24 Hydrocarbon, or cycloalkyl, phenyl, heterocycle.

15. The amino lipid compound according to claim 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-; Preferably, each of Z7 and Z8 is -(C=O)O-; preferably, (C=O) in Z7 is connected to A6, and (C=O) in Z8 is connected to A7; Alternatively, each of Z7 and Z8 is -O(C=O)-; preferably, the (C=O) in Z7 is connected to R4, and the (C=O) in Z8 is connected to R5.

16. The amino lipid compound according to any one of claims 14 or 15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: A3, A4, A6 and A7 are each independently C1-C 12 alkylene; Preferably, A3, A4, A6 and A7 are each independently C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 alkylene; Preferably, A3 is a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 alkylene; Preferably, A3 is a linear C1, C2, C3, C4 or C5 alkylene group; Preferably, A3 is a linear C2, C3 or C4 alkylene group, such as -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-; and / or Preferably, A4 is a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 alkylene; Preferably, A4 is a linear C1, C2, C3 or C4 alkylene group, such as -CH2-, -CH2CH2-, or -CH2CH2CH2-; and / or Preferably, A6 and A7 are each independently a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 alkylene; Preferably, A6 and A7 are each independently a linear C6, C7, C8, C9 or C 10 Alkylene, for example -(CH2)6-, -(CH2)7-, -(CH2)8- or -(CH2)9-.

17. The amino lipid compound according to any one of claims 14 to 16, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R1 and R2 are each independently C1-C 18 alkyl; Preferably, R1 and R2 are each independently C1-C6 alkyl; Preferably, R1 and R2 are each independently methyl, ethyl, n-propyl or isopropyl; Preferably, R1 and R2 are each independently C1-C4 alkyl, such as C1, C2, C3 or C4 alkyl; Preferably, R1 and R2 are each independently methyl; or R1 and R2, together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclic ring; Preferably, R1 and R2, together with the nitrogen atom to which they are attached, form a 5-6 membered heterocyclic ring.

18. The amino lipid compound according to any one of claims 14 to 17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R3 is C1-C 18 alkyl; Preferably, R3 is C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 or C 12 alkyl; Preferably, R3 is a linear C2, C3, C4, C5, C6, C7, C8, C9 or C 10 alkyl; Preferably, R3 is a linear C4, C5, C6, C7 or C8 alkyl group, such as n-butyl, n-pentyl, n-hexyl, n-heptyl or n-octyl.

19. The amino lipid compound according to any one of claims 14 to 18, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently C1-C 20 hydrocarbon group; Preferably, R4 and R5 are each independently C1-C 20 alkyl; Preferably, R4 and R5 are each independently a branched C3-C 20 alkyl; Preferably, R4 and R5 are each independently a branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13 alkyl; Preferably, R4 is a branched alkyl group whose branching occurs at the ɑ, β or γ position relative to Z7; R5 is a branched alkyl group whose branching occurs at the ɑ, β or γ position relative to Z8; Preferably, R4 is a branched alkyl group whose branching occurs at the ɑ or β position relative to Z7; R5 is a branched alkyl group whose branching occurs at the ɑ or β position relative to Z8; Preferably, R4 and R5 are each independently one of the following structures:

20. The amino lipid compound according to any one of claims 14 to 19, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures:

21. The amino lipid compound according to any one of claims 14 to 20, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently 22. The amino lipid compound according to any one of claims 14 to 21, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: Z7 and Z8 are each independently -(C=O)O- or -O(C=O)-; A3 is a straight-chain C1-C5 alkylene group; A4 is a straight-chain C1-C4 alkylene group; A6 and A7 are each independently C5-C 11 alkylene; R1 and R2 are each independently C1-C4 alkyl; R3 is a straight chain C2-C 10 Alkyl; and R4 and R5 are each independently C1-C 24 alkyl.

23. The amino lipid compound according to claim 22, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: Each of Z7 and Z8 is -(C=O)O-, and (C=O) in Z7 is connected to A6, and (C=O) in Z8 is connected to A7; or each of Z7 and Z8 is -O(C=O)-, and (C=O) in Z7 is connected to R4, and (C=O) in Z8 is connected to R5; and / or A3 is a straight-chain C1-C4 alkylene group; Preferably, A3 is a linear C2-C4 alkylene group, such as a linear C2, C3 or C4 alkylene group; Preferably, A3 is -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-; and / or A4 is a linear C1-C3 alkylene group, such as a linear C1, C2 or C3 alkylene group; Preferably, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-; and / or Preferably, A6 and A7 are each independently a linear C5-C 11 alkylene; Preferably, A6 and A7 are each independently a linear C6-C 10 Alkylene, such as linear C6, C7, C8, C9 or C 10 alkylene; Preferably, A6 and A7 are each independently a linear C7, C8 or C9 alkylene group, such as -(CH2)7-, -(CH2)8- or -(CH2)9-; and / or R1 and R2 are each independently C1, C2, C3 or C4 alkyl; Preferably, R1 and R2 are each independently methyl, ethyl, n-propyl or isopropyl; Preferably, R1 and R2 are each independently methyl; and / or R3 is a linear C4-C8 alkyl group, such as a C4, C5, C6, C7 or C8 alkyl group; Preferably, R3 is n-butyl, n-pentyl, n-hexyl, n-heptyl or n-octyl; and / or R4 and R5 are each independently a branched C3-C 20 Alkyl groups, such as branched C5-C 20 Alkyl, branched C7-C 19 Alkyl, branched C9-C 18 Alkyl, branched C 10 -C 18 Alkyl, branched C3-C 10 Alkyl, branched C 11 -C 17 Alkyl, or branched C 18 -C 20 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 17 Alkyl groups, such as branched C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 alkyl; Preferably, R4 and R5 are each independently a branched C 11 -C 13 Alkyl groups, such as branched C 11 、C 12 、C 13 alkyl; Preferably, R4 is a branched C3-C 20 Alkyl, relative to Z7, α, β or γ position; R5 is a branched C3-C 20 Alkyl, branching occurs at the α, β or γ position relative to Z8; Preferably, R4 is a branched C3-C 20 Alkyl, branching occurs at the α or β position relative to Z7; R5 is a branched C3-C 20 Alkyl, branching occurs at the α or β position relative to Z8; Preferably, R4 and R5 are each independently one of the following structures: Preferably, R4 and R5 are each independently one of the following structures: Preferably, R4 and R5 are each independently 24. The amino lipid compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the amino lipid compound has one of the structures shown below:

25. Lipid nanoparticles comprising the amino lipid compound according to any one of claims 1 to 24.

26. The lipid nanoparticle according to claim 25, further comprising one or more of a helper lipid, a structural lipid, and a PEG-lipid; Furthermore, the lipid nanoparticles further contain the auxiliary lipid, the structural lipid and the PEG-lipid.

27. The lipid nanoparticle of claim 26, wherein the lipid nanoparticle comprises, based on the total amount of the amino lipid compound according to any one of claims 1 to 24, the helper lipid, the structural lipid, and the PEG-lipid: An amount (molar percentage) of the amino lipid compound of any one of claims 1 to 24 of about 25.0%-75.0%, for example, about 25.0%-28.0%, 28.0%-32.0%, 32.0%-35.0%, 35.0%-40.0%, 40.0%-42.0%, 42.0%-45.0%, 45.0%-48.0%, 48.0%-55.0%, 55.0%-65.0%, 65.0%-75.0%, 45.0%-46.3%, 46.3%-48.0%, 48.0%-49.5%, 49.5%-50.0%, 50.0%-55.0% or 60.0%-65.0%; and / or The helper lipid is present in an amount (molar percentage) of about 5.0% to 45.0%, for example, about 5.0% to 10.0%, 10.0% to 16.0%, 16.0% to 25.0%, 25.0% to 33.5%, 33.5% to 37.0%, 37.0% to 40.0%, 40.0% to 42.0%, 42.0% to 45.0%, 5.0% to 9.0%, 9.0% to 9.4%, 9.4% to 10.0%, 10.0% to 10.5%, 10.5% to 11.0%, 11.0% to 15.0%, 15.0% to 16.0%, 16.0% to 18.0%, 18.0% to 20.0%, or 20.0% to 25.0%; and / or The following amount (molar percentage) of the structural lipid: about 0.0%-50.0%, for example, about 0.0%-10.0%, 10.0%-15.5%, 15.5%-22.5%, 22.5%-35.0%, 35.0%-36.5%, 36.5%-39.5%, 39.5%-40.5%, 40.5%-41.5%, 41.5%-45.0%, 45.0%-46.5%, 46.5%-50.0%, 15.5%-18.5%, 18.5%-22.5%, 22.5%-23.5%, 23.5% -28.5%, 28.5%-33.5%, 33.5%-35.0%, 36.5%-38.0%, 38.0%-38.5%, 38.5%-39.0%, 39.0%-39.5%, 41.5%-42.5%, 42.5%-42.7%, 42.7%-43.0%, 43.0%-43.5%, 43.5%-45.0%, or 46.5%-48.5%; or, about 50.0%-55.0%, such as about 50.0%-51.5%, 51.5%-53.5%, or 53.5%-55.0%. And / or The PEG-lipid is present in an amount (molar percentage) of about 0.5%-5.0%, for example, about 0.5%-1.0%, 1.0%-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, 4.5%-5.0%, 1.5%-1.6% or 1.6%-2.0%.

28. The lipid nanoparticle according to any one of claims 26 or 27, wherein: The helper lipid is a phospholipid substance; Preferably, the helper lipid is one or more selected from DSPC, DOPE, DOPC, DOPS, DSPG, DPPG, DPPC, DGTS and lysophospholipids; Preferably, the helper lipid is one or more selected from DSPC, DOPE, DOPC and DOPS; Preferably, the helper lipid is DSPC and / or DOPE; and / or The structural lipids are sterol substances; Preferably, the structured lipid is at least one selected from cholesterol, cholesterol ester, steroid hormones, steroid vitamins, bile acid, cholesterol, ergosterol, β-sitosterol and oxidized cholesterol derivatives; Preferably, the structured lipid is at least one selected from cholesterol, cholesterol ester, steroid hormones, steroid vitamins and bile acid; Preferably, the structural lipid is cholesterol, even more preferably high-purity cholesterol, particularly injection-grade high-purity cholesterol, such as CHO-HP; and / or The PEG-lipid is a conjugate of polyethylene glycol and a lipid structure; Preferably, the PEG-lipid is selected from PEG-DMG and PEG-DSPE, preferably PEG-DMG; Preferably, the PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristyl glycerol; Preferably, the average molecular weight of the PEG is about 2000 to 5000, preferably about 2000, such as PEG2000-DMG.

29. The lipid nanoparticle of any one of claims 26 to 28, wherein the molar ratio of the amino lipid compound according to any one of claims 1 to 24: helper lipid: structural lipid: PEG-lipid is about 45:11:41.5:2.5, or 42.0:10.5:45.0:2.5, or 42.0:16.0:39.5:2.5, or 40.0:16.0:41.5:2.5, or 40.0:18.0:39.5:2.

5. .5:2.5, or 35.0:16.0:46.5:2.5, or 35.0:25.0:36.5:3.5, or 28.0:33.5:35.0:3.5, or 32.0:37.0:40.5:0.5, or 35.0:40.0:22.5:2.5, or 40.0:42.0:15.5:2.5, or 45:10:42.5:2.5, or 40.0:20.0:38.5:1.5, or 45 .0:15.0:38.5:1.5、or 55.0:5.0:38.5:1.5、or 60.0:5.0:33.5:1.5、or 45.0:20.0:33.5:1.5、or 50.0:20.0:28.5:1.5、or 55.0:20.0:23.5:1.5、or 60.0:20.0:18.5:1.5、or 40.0:15.0:43.5:1.5、or 50.0:15.0: 33.5:1.5, or 55.0:15.0:28.5:1.5, or 60.0:15.0:23.5:1.5, or 40.0:10.0:48.5:1.5, or 45.0:10.0:43.5:1.5, or 55.0:10.0:33.5:1.5, or 40.0:5.0:53.5:1.5, or 45.0:5.0:48.5:1.5, or 50.0:5.0:43.5:1.5; Preferably, the helper lipid is DOPE, and the structural lipid is CHO-HP.

30. The lipid nanoparticle of any one of claims 26 to 28, wherein the molar ratio of the amino lipid compound according to any one of claims 1 to 24: helper lipid: structural lipid: PEG-lipid is about 48.0:10.0:40.5:1.5, or 50.0:10.0:38.5:1.5, or 50.0:9.0:38.0:3.0, or 49.5:10.0:39.0:1.5, or 46.3:9.4:

42. .7:1.6、or 45.0:9.0:43.0:3.0、or 45.0:11.0:41.5:2.5、or 42.0:10.5:45.0:2.5、or 42.0:16.0:39.5:2.5、or 40.0:16.0:41.5:2.5、or 40.0:18.0:39.5:2.5、or 35.0:40.0:22.5:2.5、or 40.0:20.0:38.5:1.5、or 45.0:15.0:38.5:1.5, or 55.0:5.0:38.5:1.5, or 60.0:5.0:33.5:1.5, or 45.0:20.0:33.5:1.5, or 50.0:20.0:28.5:1.5, or 55.0:20.0:23.5:1.5, or 60.0:20.0:18.5:1.5, or 40.0:15.0:43.5:1.5, or 50.0:15.0 :33.5:1.5, or 55.0:15.0:28.5:1.5, or 60.0:15.0:23.5:1.5, or 40.0:10.0:48.5:1.5, or 45.0:10.0:43.5:1.5, or 55.0:10.0:33.5:1.5, or 40.0:5.0:53.5:1.5, or 45.0:5.0:48.5:1.5, or 50.0:5.0:43.5:1.5; Preferably, the helper lipid is DSPC and the structural lipid is CHO-HP.

31. The lipid nanoparticle according to any one of claims 25 to 30, further comprising a nucleic acid; Preferably, the mass ratio of the amino lipid compound according to any one of claims 1 to 24 and the nucleic acid is about (5-30):1, such as about (5-10):1, (10-15):1, (15-20):1, (20-25):1 or (25-30):1, preferably about 10:1; and / or Preferably, the nucleic acid is selected from RNA, antisense oligonucleotide, DNA; Preferably, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single-stranded guide RNA (sgRNA), cas9 mRNA, or a mixture thereof; and / or Preferably, the DNA is a plasmid.

32. A composition comprising the amino lipid compound according to any one of claims 1 to 24 or the lipid nanoparticle according to any one of claims 25 to 31, and a pharmaceutically acceptable carrier, diluent or excipient.

33. The composition of claim 32, wherein: The composition further comprises a buffer; Preferably, the buffer is selected from phosphate buffer and Tris buffer, preferably phosphate buffer; and / or Preferably, the concentration of the buffer is about 5 mmol / L-30 mmol / L, preferably about 10 mmol / L; and / or Preferably, the pH of the buffer is about 6-8, preferably about 7-8, more preferably about 7-7.5; and / or The composition further comprises a cryoprotectant; Preferably, the cryoprotectant is selected from sucrose and trehalose, preferably sucrose; and / or Preferably, the concentration of the cryoprotectant is about 50 mg / ml-100 mg / ml.

34. The composition according to any one of claims 32 or 33, which is a pharmaceutical composition.

35. Use of a compound according to any one of claims 1 to 24 for preparing a carrier for an active ingredient; Preferably, wherein: The carrier is a lipid nanoparticle according to any one of claims 25 to 31; and / or The active ingredient is a pharmaceutical active ingredient, preferably a nucleic acid; and / or Preferably, the nucleic acid is selected from RNA, antisense oligonucleotide, DNA; Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA); and / or Preferably, the DNA is a plasmid.

36. Use of a compound according to any one of claims 1 to 24, a lipid nanoparticle according to any one of claims 25 to 31 or a composition according to any one of claims 32 to 34 in the preparation of a medicament, Preferably, the drug is used for gene therapy, gene vaccination, protein replacement therapy, antisense therapy or treatment by interfering RNA; Further preferably, the gene therapy is used to treat cancer and genetic diseases, wherein the cancer is preferably one or more selected from lung cancer, gastric cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, blood cancer or prostate cancer; and / or the genetic disease is preferably one or more selected from hemophilia, thalassemia and Gaucher's disease; and / or The genetic vaccination is used to treat cancer, allergy, toxicity and pathogen infection, wherein the pathogen is preferably selected from one or more of viruses, bacteria or fungi.

37. Use of the compound according to any one of claims 1 to 24, the lipid nanoparticle according to any one of claims 25 to 31, or the composition according to any one of claims 32 to 34 in the preparation of a medicament for nucleic acid transfer; Preferably, the nucleic acid is selected from RNA, antisense oligonucleotide, DNA; Preferably, the RNA is selected from the group consisting of mRNA, ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small inhibitory RNA (siRNA) and small nuclear RNA (snRNA); and / or Preferably, the DNA is a plasmid.

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