Aminolipid compounds, methods for preparing the same, compositions thereof, and their uses

Abstract

Amino lipid compounds, methods for preparing the same, compositions thereof, and uses thereof. Specifically, amino lipid compounds represented by formula (I), or pharmaceutically acceptable salts or stereoisomers thereof, and their use in preparing lipid nanoparticles for delivering active ingredients, as well as compositions containing amino lipid compounds, particularly lipid nanoparticles, and their use are disclosed. JPEG2025523019000124.jpg24149

Description

Detailed description of the invention

[0001] [Technical field] This disclosure relates to aminolipid compounds that can be used to prepare lipid nanoparticles for delivering active ingredients, and to methods for preparing the same. This disclosure also relates to compositions containing aminolipid compounds, particularly lipid nanoparticles, and their use.

[0002] [Background technology] Gene therapy agents deliver genes containing specific genetic information to target cells through artificial means, and the expressed target protein interferes with or modulates the expression of the relevant gene to achieve a clinical therapeutic effect. However, directly introducing naked nucleic acids into cells is difficult. External force or a vector is necessary to achieve gene delivery. However, differences in genetic and cellular structure necessitate the matching of lipid particles with different properties. Therefore, to meet the requirements for gene therapy agent delivery, it is necessary to develop different vectors or adjuvants, particularly aminolipid compounds that can be used to deliver nucleic acid active components, as well as related preparation methods and uses.

[0003] [Overview of the prefecture] One aspect of this disclosure provides an aminolipid compound represented by formula (I): [ka] (In the formula, R1, R2, R3, R4, R5, Z1, Z2, Z3, Z4, Z5, Z6, Z7, Z8, A1, A2, A3, A4, A5, A6, and A7 are defined as follows):

[0004] Another aspect of this disclosure provides a method for preparing aminolipid compounds.

[0005] Another aspect of this disclosure provides the use of aminolipid compounds in the production of vehicles for active ingredients.

[0006] Another aspect of this disclosure provides lipid nanoparticles comprising an aminolipid compound.

[0007] Another aspect of this disclosure provides a composition comprising an aminolipid compound.

[0008] Another aspect of this disclosure provides the use of aminolipid compounds, lipid nanoparticles, or compositions in the manufacture of pharmaceuticals.

[0009] Another aspect of this disclosure provides the use of aminolipid compounds, lipid nanoparticles, or compositions in the manufacture of pharmaceuticals for nucleic acid transfer.

[0010] [Modes for carrying out the invention] definition Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by those skilled in the art. References to technology used herein are intended to mean technology commonly understood in the art, including modifications or substitutions of equivalent technology that would be obvious to those skilled in the art. The following terms are considered to be well understood by those skilled in the art, but the following definitions are provided for the better explanation of this disclosure.

[0011] As used herein, the terms “comprising,” “including,” “having,” “containing,” or “accompanying,” and other variations thereof, are comprehensive or open-ended and do not exclude other unenumerated elements or method steps.

[0012] As used herein, the term "hydrocarbyl" refers to a group that remains after one hydrogen atom is lost from an aliphatic hydrocarbon, including linear or branched saturated or unsaturated hydrocarbyl groups. Hydrocarbyl groups include alkyl, alkenyl, and alkynyl groups. Preferably, the hydrocarbyl group has 1 to 24 carbon atoms (C1-C 24(Hydrocarbyl), 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 (Hydrocarbyl). Examples of hydrocarbyl groups include C1-C 20 hydrocarbyl, C1-C 18 hydrocarbyl, C1-C 16 hydrocarbyl, C1-C 12 hydrocarbyl, C1-C 10 hydrocarbyl, C1-C8 hydrocarbyl, C1-C7 hydrocarbyl, C1-C6 hydrocarbyl, C1-C4 hydrocarbyl, C1-C3 hydrocarbyl, C1-C2 hydrocarbyl, C2-C8 hydrocarbyl, C2-C4 hydrocarbyl, C4-C8 hydrocarbyl, C4-C9 hydrocarbyl, C5-C8 hydrocarbyl, C1-C4 hydrocarbyl, C2-C8 hydrocarbyl, C3 hydrocarbyl, C4 hydrocarbyl, C5 hydrocarbyl, C6 hydrocarbyl, C7 hydrocarbyl and C8 hydrocarbyl, but are not limited thereto.

[0013] As used herein, the term "hydrocarbylene" refers to a divalent group remaining after one hydrogen atom is further lost from the hydrocarbyl defined above.

[0014] As used herein, the term "alkyl" is a straight-chain or branched saturated monovalent hydrocarbyl. Preferably, the alkyl group has 1 to 24 carbon atoms (C1-C<0OO0011>​​​​​​​​​​​​Alkyl, C1-C 16 Alkyl, C1-C 12 Alkyl, C1-C 10 Examples include, but are not limited to, 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-tridecyl, and tridecan-7-yl.

[0015] As used herein, the term "alkylene" refers to the divalent group that remains after one more hydrogen atom has been lost from the alkyl group defined above.

[0016] As used herein, the term "alkenyl" refers to a monovalent hydrocarbyl, either linear or branched, containing one or more double bonds (C=C). Preferably, the alkenyl group has 2 to 24 carbon atoms (C2-C). 24 Alkenyls) 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 groups have 1, 2, 3, 4 or more double bonds. An alkenyl group is a C2-C group having 1, 2, 3, 4 or more double bonds. 20 Alkenyl, C2-C 18 Alkenyl, C2-C 16 Alkenyl, C2-C 12 Alkenyl, C2-C 10Examples include, but are not limited to, alkenyls, C2-C8 alkenyls, C2-C7 alkenyls, C2-C6 alkenyls, C2-C4 alkenyls, C2-C3 alkenyls, C4-C8 alkenyls, C4-C9 alkenyls, and C5-C8 alkenyls. Some more specific examples include, but are not limited to, vinyl, propenyl, buta-1-enyl, buta-2-enyl, penta-1-enyl, penta-2-enyl, hexa-1-enyl, hexa-2-enyl, hexa-3-enyl, hepta-1-enyl, hepta-2-enyl, hepta-3-enyl, octa-1-enyl, octa-2-enyl, octa-3-enyl, nona-1-enyl, nona-2-enyl, and nona-3-enyl. In some preferred embodiments, the alkenyl group has one double bond.

[0017] As used herein, the term "alkenylene" refers to the divalent group that remains after one more hydrogen atom has been lost from the alkenyl as defined above.

[0018] As used herein, the term "alkynyl" refers to a linear or branched monovalent hydrocarbyl group containing one or more triple bonds (C≡C). Preferably, the alkynyl group has 2 to 24 carbon atoms (C2-C). 24 Alkynnyls, for example, have 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 groups have 1, 2, 3, 4 or more triple bonds. As alkynyl groups, C2-C groups have 1, 2, 3, 4 or more triple bonds. 20 Alkinyl, C2-C 18 Alkinyl, C2-C 16 Alkinyl, C2-C 12 Alkinyl, C2-C 10Examples include, but are not limited to, alkynyl, C2-C8 alkynyl, C2-C7 alkynyl, C2-C6 alkynyl, C2-C4 alkynyl, C2-C3 alkynyl, C4-C8 alkynyl, C4-C9 alkynyl, and C5-C8 alkynyl. Some more specific examples include, but are not limited to, ethinyl, propynyl, buta-1-inyl, buta-2-inyl, penta-1-inyl, penta-2-inyl, hexa-1-inyl, hexa-2-inyl, hexa-3-inyl, hepta-1-inyl, hepta-2-inyl, hepta-3-inyl, octa-1-inyl, octa-2-inyl, octa-3-inyl, nona-1-inyl, nona-2-inyl, and nona-3-inyl. In some preferred embodiments, the alkynyl group has one triple bond.

[0019] As used herein, the term "alkynylene" refers to the divalent group that remains after one more hydrogen atom is lost from the alkynyl as defined above.

[0020] As used herein, the terms “cyclohydrocarbyl,” “cyclohydrocarbylene,” 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) in the ring) monocyclic or polycyclic hydrocarbon rings having, for example, 3 to 10 (preferably 3 to 8, more preferably 3 to 6, e.g., 5 to 6 or 5 to 7) ring carbon atoms, including, but not limited to, cyclopropyl(ene)(ring), cyclobutyl(ene)(ring), cyclopentyl(ene)(ring), cyclohexyl(ene)(ring), cycloheptyl(ene)(ring), cyclooctyl(ene)(ring), cyclononyl(ene)(ring), and cyclohexenyl(ene)(ring).

[0021] As used herein, the term "cycloalkyl" refers to saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon rings (e.g., monocyclic rings such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclononyl, or bicyclic rings including spirocyclic, fused, or bridging systems such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]nonyl, and decalin). Cycloalkyls have 3 to 10 carbon atoms, for example, 3 to 7, 5 to 6, or 5 to 7.

[0022] As used herein, the terms “heterocycle,” “heterocyclyl,” or “heterocyclylene” mean 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, and 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, and piperazine, preferably pyrrolidine, piperidine, piperazine, and morpholine. The heterocycle may be optionally substituted with one or more substituents, for which the definition of "optionally substituted" below applies.

[0023] As used herein, the term "aryl" refers to a monocyclic or polycyclic aromatic group with a conjugated π-electron system. For example, as used herein, "C 6-14The term "aryl" refers to an aromatic group containing 6 to 14 (e.g., 6 to 12) carbon atoms, such as phenyl or naphthyl. The aryl group is optionally substituted with one or more (e.g., 1 to 3) preferred substituents.

[0024] As used herein, the term “heteroaryl” means a monocyclic or polycyclic (e.g., bicyclic or tricyclic) aromatic group having a conjugated π-electron system, wherein the ring atoms consist of carbon atoms and at least one heteroatom, and the group has 5 to 14 ring atoms (e.g., 5, 6, 7, 8, 9, 10, 11, 12, or 14) including, for example, 1, 2, 3, 4, or 13 carbon atoms and 1, 2, 3, 4, or 5 identical or different heteroatoms independently selected from N, O, S, and S(O)2. One or more ring carbon atoms in the heteroaryl group may be substituted with C(O). The heteroaryl group may be benzo-condensed. The heteroaryl group may be optionally substituted with one or more suitable substituents.

[0025] As used herein, the term “optionally substituted” means that one or more hydrogen atoms bonded to an atom or group are independently unsubstituted or substituted with one or more (e.g., 1, 2, 3, or 4) substituents. Substituents can independently be 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- to 8-membered heterocyclyl), heteroaryl, aryl C1-C6 alkyl-, heteroaryl C1-C6 alkyl, C1-C6 haloalkyl, -OC1-C6 alkyl (preferably -OC1-C 3 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-C6alkyl)C(O)(phenyl), -C(O)C1-C6alkyl, -C(O)heteroaryl (preferably -C(O)-5~7 member heteroaryl), -C(O)C1-C6alkylphenyl, -C(O)C1-C6 haloalkyl, -OC(O)C1-C6alkyl (preferably -OC(O)C1-C3alkyl), -S(O)2-C1-C6alkyl, -S(O)-C1-C6alkyl, -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), where each of alkyl, cycloalkyl, phenyl, aryl, heterocyclyl, and heteroaryl is optionally 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, nitrate The atom or group is further substituted with one or more substituents selected from -C(O)-OH, -C(O)OC1-C6alkyl, -CON(C1-C6alkyl)2, -CONH(C1-C6alkyl), -CONH2, -NHC(O)(C1-C6alkyl), -NH(C1-C6alkyl)C(O)(C1-C6alkyl), -SO2(C1-C6alkyl), -SO2(phenyl), -SO2(C1-C6 haloalkyl), -SO2NH2, -SO2NH(C1-C6alkyl), -SO2NH(phenyl), -NHSO2(C1-C6alkyl), -NHSO2(phenyl), and -NHSO2(C1-C6 haloalkyl). If an atom or group is substituted with multiple substituents, the substituents may be the same or different.

[0026] In certain embodiments, substituents include halogens (such as chlorine, bromine, fluorine, or iodine), deuterium (D), tritium (T), carboxylic acids (such as -C(=O)-OH), oxygen (such as =O), sulfur (such as =S), hydroxyl (such as -OH), ester groups (such as -C(=O)ORiii or -OC(=O)Riii), aldehyde groups (such as -C(=O)H), carbonyls (such as -C(=O)Riii, or represented by C=O), acyl halides (e.g., -C(=O)X (wherein X is selected from bromine, fluorine, chlorine, or iodine)), carbonate ester groups (such as -OC(=O)ORiii), alkoxys (such as -ORiii), acetals (e.g., -C(ORiii)2Riii (wherein each ORiii is the same or different and is an alkoxy group)), and phosphates (P(=O)43- (etc.), thiols (-SH etc.), sulfoxides (-S(=O)Riii etc.), sulfinic acid (-S(=O)OH etc.), sulfonic acid (-S(=O)2OH etc.), thioaldehydes (-C(=S)H etc.), sulfates (S(=O)4 2-(etc.), sulfonyl (-S(=O)2Riii, etc.), sulfinyl (-S(=O)Riii, etc.), amide (-C(=O)N(Riii)2 or -N(Riii)C(=O)Riii, etc.), azide (-N3, etc.), nitro (-NO2, etc.), cyano (-CN, etc.), isocyano (-NC, etc.), acyloxy (-OC(=O)Riii, etc.), amino (-N(Riii)2, -N(Riii)H or -NH2, etc.), carbamoyl (-OC(=O)N(Riii)2, -OC(=O)N(Riii)H or -OC(=O)NH2, etc.), s Lufonamides (-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, etc.), alkyl, alkenyl, alkynyl, cyclohydrocarbyl (cycloalkyl, cycloalkenyl or cycloalkynyl, etc.), heterocyclohydrocarbyl (heterocycloalkyl containing one or more heteroatoms selected from S, N and O, or selected from S, N and O) Heterocycloalkenyls containing one or more heteroatoms, aryls (phenyl or fused ring groups, etc.), heteroaryls (bicyclic heteroaryls of 8-10 membered rings containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, etc.), -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, but are not limited thereto.In all of the above, Riii is hydrogen, or an alkyl, alkenyl, or alkynyl, or heteroalkyl, heteroalkenyl, or heteroalkynyl as defined herein. In some embodiments, Riii is hydrogen, or a C1-C as defined herein. 12 Alkyl, or C1-C 12 Alkenyl, or C1-C 12 Alkinyl, or C1-C 12 Heteroalkyl, or C1-C 12 Heteroalkenyl, or C1-C 12 It is a heteroalkynyl.

[0027] In certain embodiments, the substituent itself may be further substituted with one or more substituents as defined herein, for example. For instance, a C1-C6 alkyl substituent may be further substituted with one or more substituents as defined herein.

[0028] In certain embodiments, the hydrocarbyl, hydrocarbylene, alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, cyclohydrocarbyl, cyclohydrocarbylene, hydrocarbon ring, cycloalkyl, heterocycle, heterocyclyl, heterocyclylene, aryl, and heteroaryl groups described herein may be optionally substituted with one or more substituents.

[0029] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0030] As used herein, the "C(=O)" and "(C=O)" groups are interchangeable. [ka] This represents a -(C=O)O- group. [ka] It represents.

[0031] As used herein, the term “compound” encompasses all compounds that are isotope-labeled by substituting one or more atoms with atoms having different atomic weights or mass numbers. “Isotopes” refer to atoms that have the same atomic number but different mass numbers due to a difference in the number of neutrons in their nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0032] Numerical ranges described herein should be understood to include all subranges contained within them. For example, the range "1 to 10" should be understood to include not only the explicitly listed values ​​of 1 and 10, but also any individual values ​​within the range of 1 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, and 9) as well as any 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.). This principle also applies to ranges that use only one value as the minimum or maximum value.

[0033] As used herein, “pharmaceutically acceptable salt” means an acid-addition salt or base-addition salt of a compound of the Disclosure that retains the biological efficacy and properties of the compound and is typically not biologically or otherwise undesirable. In many cases, the compounds of the present invention can form acid and / or base salts in the presence of an amino and / or carboxyl group or similar group.

[0034] pharmaceutically acceptable acid addition salts can be formed from the compounds of this disclosure as well as inorganic and / or organic acids, the inorganic acids being, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid, and the organic acids being, 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, camphanic acid, camphor-10-sulfonic acid, capric acid, hexanoic acid, octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, gluconic acid These include coheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinic 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.

[0035] Pharmaceutically acceptable base addition salts can be formed from the compounds of this disclosure as well as from inorganic and / or organic bases. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Examples of salts derived from organic bases include, but are not limited to, salts of primary amines, secondary amines, 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, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydravamin, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0036] The compounds of this disclosure or their pharmaceutically acceptable salts may contain one or more chiral centers, thus giving rise to enantiomers, diastereomers, and other stereoisomeric forms, such as amino acids, which may be defined as (R)- or (S)-, or (D)- or (L)- based on absolute stereochemistry. The present invention is intended to include all these possible isomers as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or by resolution by 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 racemics (or racemics of salts or derivatives) using, for example, chiral high-pressure liquid chromatography (HPLC). Where the compounds described herein contain an olefinic double bond or other geometrically asymmetric centers, unless otherwise specified, the compounds include E- and Z-geometric isomers. Similarly, all tautomers are intended to be considered.

[0037] As used herein, the term “stereoisomer” refers to a compound consisting of the same atoms that are bonded together via the same bonds but have different three-dimensional structures that are not interchangeable. “Enantiomers” are a pair of stereoisomers that are mirror images of each other but do not overlap. A mixture of any ratio of a pair of enantiomers may be called a “racemic” mixture. “Diastereomers” are stereoisomers that have at least two chiral atoms and are not mirror images of each other.

[0038] "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.

[0039] A "tautomer" refers to an isomer of a compound that is in equilibrium with it. The concentration of each isomer depends on the environment in which the compound is found, and can vary depending on whether the compound is solid, organic, or in an aqueous solution.

[0040] Aminolipid compounds In one embodiment, the present disclosure relates to an aminolipid compound represented by the following formula (I): [ka] or provide a pharmaceutically acceptable salt thereof or its stereoisomer, During the ceremony, 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 a -C(=O)- or bond, Z4 is -O- or -CH(OH)-, A1, A2, A3, A4, A5, A6, and A7 are each independently C1-C 12 Hydrocarbylene, cyclohydrocarbyl, phenyl, heterocyclic or bonded, R1 and R2 are independently H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, or R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocyclic ring. R3 is H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, R4 and R5 are independently C1-C 24 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, Preferably, A5, Z5, and Z6 are combined.

[0041] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in which A1 and A2 are each independently a C1-C8 hydrocarbilene or bond, e.g., a C1-C6 hydrocarbilene or bond, or a C1-C4 hydrocarbilene or bond. In some embodiments, A1 and A2 are each independently a C1-C8 alkylene, a C2-C8 alkenylene, or a C2-C8 alkylene. In some embodiments, A1 and A2 are each independently a C1-C8 alkylene, e.g., a C1-C6 alkylene, a C1-C4 alkylene, a C2-C4 alkylene, or a C1-C3 alkylene. In some embodiments, both A1 and A2 are bonds.

[0042] In some embodiments, this disclosure states that A3 is C1-C 12 The present invention provides an aminolipid compound of formula (I) above, which is a hydrocarbylene or a linkage, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. In some embodiments, A3 is a linear C1-C 12 It is hydrocarbylene. In some embodiments, A3 is a linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 It is an alkylene. In some embodiments, A3 is a linear C1-C5 alkylene, for example, a linear C1, C2, C3, C4 or C5 alkylene. In some embodiments, A3 is CH2CH2-, -CH2CH2CH2- or -CH2CH2CH2CH2-.

[0043] In some embodiments, this disclosure states that A4 is C1-C 12 The present invention provides an aminolipid compound of formula (I) above, which is a hydrocarbylene or a linkage, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. In some embodiments, A4 is a linear C1-C 12 It is hydrocarbylene. In some embodiments, A4 is a linear C1-C 12 Alkylene, C2-C 12Alkenylene or C2-C 12 It is an alkylene. In some embodiments, A4 is a linear C1-C7 alkylene, for example, a linear C1, C2, C3, C4, C5, C6, or C7 alkylene. In some embodiments, A4 is a linear C1-C3 alkylene, for example, a linear C1, C2, or C3 alkylene. In some embodiments, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0044] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in which A5 is a C1-C8 hydrocarbilene or bond, e.g., a C1-C6 hydrocarbilene or bond, a C1-C4 hydrocarbilene or bond, or a C1-C3 hydrocarbilene or bond. In some embodiments, A5 is a C1-C8 alkylene, a C2-C8 alkenylene, or a C2-C8 alkylene. In some embodiments, A5 is a C1-C8 alkylene, e.g., a C1-C6 alkylene, a C1-C4 alkylene, a C2-C4 alkylene, or a C1-C3 alkylene. In some embodiments, A5 is a bond.

[0045] In some embodiments, the Disclosure relates that A6 and A7 are independently C1-C 12 The present invention provides an aminolipid compound of formula (I) above, which is hydrocarbylene, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. In some embodiments, A6 and A7 are independently linear C1-C 12 It is hydrocarbylene. In some embodiments, A6 and A7 are each independently linear C1-C 12 Alkylene, C2-C 12 Alkenylene or C2-C 12 It is an alkynylene. In some embodiments, A6 and A7 are each independently a linear C1-C 12 Alkylenes, for example, straight-chain C2-C 12 Alkylene, C4-C 10 Alkylene, C6-C8 Alkylene, C6-C10 It is an alkylene or a C4-C8 alkylene. In some embodiments, A6 and A7 are each independently a linear C6-C 10 Alkylenes, for example, straight chain C6, C7, C8, C9 or C 10 It is an alkylene. In some embodiments, A6 and A7 are independently -(CH2)6-, -(CH2)7-, -(CH2)8-, or -(CH2)9-, respectively.

[0046] In some embodiments, R1 and R2 are independently H, C1-C 18 Hydrocarbyl, e.g., C1-C 16 Hydrocarbyl, C1-C 12 Hydrocarbyl, C1-C 10 The present disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein the compound is a hydrocarbyl, C1-C8 hydrocarbyl, C1-C6 hydrocarbyl or C1-C4 hydrocarbyl, C3-C7 cyclohydrocarbyl, e.g., C3-C6 cyclohydrocarbyl or C5-C6 cyclohydrocarbyl, phenyl, or a 3- to 7-membered heterocycle, e.g., a 4- to 6-membered heterocycle, or R1 and R2 together with the nitrogen atom to which they are bonded to form a 4- to 7-membered heterocycle, e.g., a 5- to 6-membered heterocycle.

[0047] In some embodiments, R1 and R2 are independently H, C1-C 18 Alkylene, C2-C 18 Alkenylene or C2-C 18 It is an alkynylene. In some embodiments, R1 and R2 are each independently C1-C 18 These are alkyl groups, such as C1-C4 alkyl groups, C3-C6 cycloalkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups, phenyl groups, or 5-6 membered heterocycles. In some embodiments, R1 and R2, together with the nitrogen atoms to which they are bonded, form 5-6 membered heterocycles such as pyrrolidine, piperidine, piperazine, or morpholine.

[0048] 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.

[0049] In some embodiments, the present disclosure provides the above amino lipid compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein R3 is H, C1-C 18 hydrocarbyl, such as C1-C 16 hydrocarbyl, C2-C 12 hydrocarbyl, C4-C 10 hydrocarbyl or C4-C8 hydrocarbyl, C3-C7 cyclohydrocarbyl, such as C3-C6 cyclohydrocarbyl or C5-C6 cyclohydrocarbyl, phenyl, or a 3- to {7}-membered heterocyclic ring, such as a 4- to 6-membered heterocyclic ring. In some embodiments, R3 is C1-C 18 alkyl, C2-C 18 alkenyl or C2-C 18 alkynyl. In some embodiments, R3 is C1-C 18 alkyl, 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. In some embodiments, R3 is a straight-chain C4-C8 alkyl, such as straight-chain C4, C5, C6, C7 or C8 alkyl. In some embodiments, R3 is n-butyl, n-pentyl, n-hexyl, n-heptyl or n-octyl..

[0050] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in which 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.

[0051] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z3 is -C(=O)-.

[0052] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z7 and Z8 are independently -(C=O)O-, -O(C=O)-, or a bond.

[0053] In some embodiments, the disclosure provides an aminolipid compound of formula (I) 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.

[0054] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z7 and Z8, respectively, are -(C=O)O-. In some embodiments, the (C=O) in Z7 is bonded to A6, and the (C=O) in Z8 is bonded to A7.

[0055] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z7 and Z8, respectively, are -O(C=O)-. In some embodiments, the (C=O) in Z7 is bonded to R4, and the (C=O) in Z8 is bonded to R5.

[0056] In some embodiments, the disclosure provides an aminolipid compound of formula (I) 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 bonded to A7. In some embodiments, Z7 is a bond and Z8 is -O(C=O)-. In some embodiments, the (C=O) in Z8 is bonded to R5. In some embodiments, Z8 is a bond and Z7 is -(C=O)O-. In some embodiments, the (C=O) in Z7 is bonded to A6. In some embodiments, Z8 is a bond and Z7 is -O(C=O)-. In some embodiments, the (C=O) in Z7 is bonded to R4.

[0057] In some embodiments, the present disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z7 and Z8 are each bonded.

[0058] In some embodiments, the Disclosure relates that R4 and R5 are independently C1-C 20 The present invention provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, which is a hydrocarbyl, a C3-C7 cyclohydrocarbyl, such as a C3-C6 cyclohydrocarbyl or a C5-C6 cyclohydrocarbyl, a phenyl, or a 3- to 7-membered heterocycle, such as a 4- to 6-membered heterocycle. In some embodiments, R4 and R5 are independently C1-C 20 The components are hydrocarbyl, C3-C6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, phenyl, or a 5-6 membered heterocycle.

[0059] In some embodiments, the Disclosure relates that R4 and R5 are independently C1-C 20Provided is an amino lipid compound of formula (I) that is a hydrocarbyl, or a pharmaceutically acceptable salt or stereoisomer thereof.

[0060] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) wherein R4 and R5 are each independently C1-C 20 alkyl, or a pharmaceutically acceptable salt or stereoisomer thereof.

[0061] In some embodiments, the present disclosure provides an amino lipid compound of formula (I) wherein R4 and R5 are each independently branched C3-C 20 alkyl, 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, or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, R4 and R5 are each independently branched C 11 -C 17 alkyl, such as branched C 11 , C 12 , C 13 , C 14 , C 15 , C 16 or C 17 alkyl. In some embodiments, R4 and R5 are each independently branched C 11 -C 13 alkyl, such as branched C 11 , C 12 or C 13 alkyl. In some embodiments, R4 is a branched alkyl group where the branch occurs at the α, β, or γ position relative to Z7, such as

Chemical formula

[0062] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures. [ka]

[0063] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R4 and R5 are each independently one of the following structures. [ka]

[0064] In some embodiments, this disclosure describes how R4 and R5 are independently of each other. [ka] or [ka] The present invention provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0065] In some embodiments, this disclosure is, Z3 is -C(=O)-, A4, C1-C 12 The present invention provides an aminolipid compound of formula (I) above, which is alkylene, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. In some embodiments, A4 is a linear C1-C 12 Alkylenes, for example, straight-chain C1-C 10 A4 is an alkylene, C1-C8 alkylene, C1-C6 alkylene, C1-C4 alkylene, or C1-C3 alkylene. In some embodiments, A4 is a linear C1-C4 alkylene, for example, a linear C1, C2, C3, or C4 alkylene. In some embodiments, A4 is -CH2-, -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0066] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z4 is -O-.

[0067] In some embodiments, the disclosure provides an aminolipid compound of formula (I) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z4 is -CH(OH)-.

[0068] In another embodiment, the present disclosure relates to aminolipid compounds represented by the following formulas (II) and (III): [ka] or provide a pharmaceutically acceptable salt thereof or its stereoisomer, During the ceremony, Z7 and Z8 are 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 Hydrocarbylene, cyclohydrocarbyl, phenyl, heterocyclic or bonded, R1 and R2 are independently H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, or R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocyclic ring. R3 is H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, R4 and R5 are independently C1-C 24 It is hydrocarbyl or cyclohydrocarbyl, phenyl, and heterocyclic.

[0069] In some embodiments, the disclosure provides aminolipid compounds of formulas (II) and (III) above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in which Z7 and Z8 are 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 bonded to A6, and the (C=O) in Z8 is bonded to A7. In some embodiments, each of Z7 and Z8 is -O(C=O)-. In some embodiments, the (C=O) in Z7 is bonded to R4, and the (C=O) in Z8 is bonded to R5.

[0070] In some embodiments, the Disclosure relates that A3, A4, A6, and A7 are each independently C1-C 12 The present invention provides aminolipid compounds of formulas (II) and (III) above, which are alkylenes, or pharmaceutically acceptable salts thereof or stereoisomers thereof. In some embodiments, A3, A4, A6 and A7 are, independently, C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 or C 12 It is alkylene.

[0071] In some embodiments, A3 is a linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C10 , C 11 or C 12 It is an alkylene. In 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, e.g., -CH2CH2-, -CH2CH2CH2-, or -CH2CH2CH2CH2-.

[0072] In some embodiments, A4 is a linear chain C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 or C 12 It is an alkylene. In some embodiments, A4 is a linear C1, C2, C3, or C4 alkylene, for example -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0073] In some embodiments, A6 and A7 are independently linear C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 or C 12 It is an alkylene. In some embodiments, A6 and A7 are independently linear C6, C7, C8, C9 or C 10 These are alkylenes, such as -(CH2)6-, -(CH2)7-, -(CH2)8-, or -(CH2)9-.

[0074] In some embodiments, the Disclosure relates that R1 and R2 are independently C1-C 18The present invention provides aminolipid compounds of the above formulas (II) and (III) that are alkyl, or pharmaceutically acceptable salts thereof, or stereoisomers thereof. In some embodiments, R1 and R2 are independently C1-C6 alkyl groups. In some embodiments, R1 and R2 are independently methyl, ethyl, n-propyl, or isopropyl groups. In some embodiments, R1 and R2 are independently C1-C4 alkyl groups, such as C1, C2, C3, or C4 alkyl groups. In some embodiments, R1 and R2 are independently methyl groups.

[0075] In some embodiments, the disclosure provides aminolipid compounds of formulas (II) and (III) above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in which R1 and R2, together with the nitrogen atoms to which they are bonded, form a 4- to 7-membered heterocycle. In some embodiments, R1 and R2, together with the nitrogen atoms to which they are bonded, form a 5- to 6-membered heterocycle.

[0076] In some embodiments, this disclosure describes R3 as C1-C 18 The present invention provides aminolipid compounds of the above formulas (II) and (III) that are alkyl, or pharmaceutically acceptable salts thereof or stereoisomers thereof. In some embodiments, R3 is C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 or C 12 It is alkyl. In some embodiments, R3 is linear C2, C3, C4, C5, C6, C7, C8, C9 or C 10 It is alkyl. In some embodiments, R3 is a linear C4, C5, C6, C7, or C8 alkyl, such as n-butyl, n-pentyl, n-hexyl, n-heptyl, or n-octyl.

[0077] In some embodiments, the Disclosure relates that R4 and R5 are independently C1-C 20The present invention provides aminolipid compounds of formulas (II) and (III) above, which are hydrocarbyl compounds, or pharmaceutically acceptable salts thereof or stereoisomers thereof.

[0078] In some embodiments, R4 and R5 are independently C1-C 20 It is alkyl.

[0079] In some embodiments, R4 and R5 are independently branched C3-C 20 Alkyl, for example, 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 It is alkyl. In some embodiments, R4 and R5 are independently branched C 11 -C 17 Alkyl, for example, branched C 11 , C 12 , C 13 , C 14 , C 15 , C 16 or C 17 It is alkyl. In some embodiments, R4 and R5 are independently branched C 11 -C 13 Alkyl, for example, branched C 11 , C 12 or C 13 It is an alkyl group. In some embodiments, R4 is a branched alkyl group in which branching occurs at the α, β, or γ position relative to Z7, and R5 is a branched alkyl group in which branching occurs at the α, β, or γ position relative to Z8. In some embodiments, R4 is a branched alkyl group in which branching occurs at the α or β position relative to Z7, and R5 is a branched alkyl group in which branching occurs at the α or β position relative to Z8. In some embodiments, R4 and R5 are each independently one of the following structures. [ka]

[0080] In some embodiments, the disclosure provides aminolipid compounds of formulas (II) and (III) above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof, in which R4 and R5 are each independently one of the following structures. [ka]

[0081] In some embodiments, this disclosure describes how R4 and R5 are independently of each other. [ka] or [ka] The present invention provides aminolipid compounds of the above formulas (II) and (III), or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0082] In some embodiments, this disclosure is, Z7 and Z8 are independently -(C=O)O- or -O(C=O)-, A3 is a linear C1-C5 alkylene, A4 is a linear C1-C4 alkylene. A6 and A7 are independently C5-C 11 It is alkylene, R1 and R2 are each independently a C1-C4 alkyl group. R3 is a linear C2-C 10 It is alkyl, R4 and R5 operate independently, C1-C 24The present invention provides alkyl aminolipid compounds of formulas (II) and (III) above, or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0083] In some embodiments, Z7 and Z8 are each -(C=O)O-, where (C=O) in Z7 is bonded to A6 and (C=O) in Z8 is bonded to A7. In some embodiments, Z7 and Z8 are each -O(C=O)-, where (C=O) in Z7 is bonded to R4 and (C=O) in Z8 is bonded to R5.

[0084] 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-.

[0085] In some embodiments, A4 is a linear C1-C3 alkylene, for example, a linear C1, C2, or C3 alkylene. In some embodiments, A4 is -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0086] In some embodiments, A6 and A7 are independently linear C5-C 11 It is an alkylene. In some embodiments, A6 and A7 are each independently a linear C6-C 10 Alkylenes, for example, straight chain C6, C7, C8, C9 or C 10 It is an alkylene. In some embodiments, A6 and A7 are each independently a linear C7, C8, or C9 alkylene, e.g., -(CH2)7-, -(CH2)8-, or -(CH2)9-.

[0087] In some embodiments, R1 and R2 are independently C1, C2, C3, or C4 alkyl groups. In some embodiments, R1 and R2 are independently methyl, ethyl, n-propyl, or isopropyl groups. In some embodiments, R1 and R2 are independently methyl groups.

[0088] 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.

[0089] In some embodiments, R4 and R5 are independently branched C3-C 20 Alkyl, for example, 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 It is alkyl. In some embodiments, R4 and R5 are independently branched C 11 -C 17 Alkyl, for example, branched C 11 , C 12 , C 13 , C 14 , C 15 , C 16 or C 17 It is alkyl. In some embodiments, R4 and R5 are independently branched C 11 -C 13 Alkyl, for example, branched C 11 , C 12 or C 13It is an alkyl group. In some embodiments, R4 is a branched alkyl group in which branching occurs at the α, β, or γ position relative to Z7, and R5 is a branched alkyl group in which branching occurs at the α, β, or γ position relative to Z8. In some embodiments, R4 is a branched alkyl group in which branching occurs at the α or β position relative to Z7, and R5 is a branched alkyl group in which branching occurs at the α or β position relative to Z8.

[0090] In some embodiments, R4 and R5 are each independently one of the following structures: [ka]

[0091] In some embodiments, R4 and R5 are each independently one of the following structures: [ka]

[0092] In some embodiments, R4 and R5 are independent of each other. [ka] or [ka] That is the case.

[0093] In various embodiments, the present disclosure provides an aminolipid compound of formula (I), (II), or (III) above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the aminolipid compound has one of the structures shown in Table 1 below. [Table 1] JPEG0007900866000021.jpg209149 JPEG0007900866000022.jpg211149 JPEG0007900866000023.jpg222149 JPEG0007900866000024.jpg223149 JPEG0007900866000025.jpg214149 JPEG0007900866000026.jpg126149

[0094] All aminolipid compounds of this disclosure possess hydrophobic properties due to the presence of long nonpolar residues and hydrophilic properties due to the amino group. This amphiphilic property allows the aminolipid compounds of this disclosure to be used to form lipid nanoparticles, such as lipid bilayers, micelles, liposomes, and the like.

[0095] In the context of this disclosure, the term “lipid nanoparticles” means nanometer-sized materials produced by introducing aminolipid compounds into an aqueous solution. The particles are, in particular, lipid nanoparticles, lipid bilayer vesicles (liposomes), multilayer vesicles, or micelles.

[0096] In some preferred embodiments, the lipid nanoparticles are liposomes containing the aminolipid compounds of the Disclosure. Within the scope of the Disclosure, liposomes are microvesicles comprising a bilayer of lipid amphiphilic molecules encapsulating an aqueous compartment.

[0097] Liposome formation is not a spontaneous process. When lipids are introduced into water, lipid vesicles are formed first, thus creating a bilayer or series of bilayers, each separated by water molecules. Liposomes can be formed by sonicating lipid vesicles in water.

[0098] In the context of this disclosure, the term “lipid bilayer” means a thin film formed by two layers of lipid molecules. The term “micelle” means an aggregate of surfactant molecules dispersed in a liquid colloid. A typical micelle in aqueous solution, upon contact with water, forms aggregates with a hydrophilic head region and chelates the hydrophobic single tail region at the center of the micelle.

[0099] In one embodiment, the present disclosure provides the use of the aminolipid compounds of the present disclosure for the production of a vehicle for an active ingredient. In some embodiments, the vehicle is in the form of lipid nanoparticles such as lipid bilayers, micelles, or liposomes.

[0100] Lipid nanoparticles In another aspect, the Disclosure provides lipid nanoparticles containing the aminolipid compounds of the Disclosure.

[0101] In some embodiments, the lipid nanoparticles further contain a pharmaceutically acceptable carrier, diluent, or excipient.

[0102] In some embodiments, the lipid nanoparticles further contain one or more of the following: helper lipids, structural lipids, and PEG-lipids (polyethylene glycol-lipids).

[0103] In some further embodiments, the lipid nanoparticles further contain helper lipids, structural lipids, and PEG-lipids.

[0104] In some embodiments, the lipid nanoparticles contain aminolipid compounds in amounts (mol percent) of approximately 25.0% to 75.0%, for example, approximately 25.0% to 28.0%, 28.0% to 32.0%, 32.0% to 35.0%, 35.0% to 40.0%, 40.0% to 42.0%, 42.0% to 45.0%, 45.0% to 48.0%, 48.0% to 55.0%, 55.0% to 65.0%, 65.0% to 75.0%, 45.0% to 46.3%, 46.3% to 48.0%, 48.0% to 49.5%, 49.5% to 50.0%, 50.0% to 55.0%, or 60.0% to 65.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids.

[0105] In some embodiments, the lipid nanoparticles are present in amounts of approximately 5.0% to 45.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids, for example, approximately 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%, and 40.0% to 42%. It contains helper lipids in amounts (mol percent) of 0.0%, 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%.

[0106] In some embodiments, lipid nanoparticles are present in amounts of approximately 0.0% to 50.0%, for example, approximately 0.0% to 10.0%, 10.0% to 15.5%, 15.5% to 22.5%, 22.5% to 35.0%, 35.0% to 36.5%, 36.5% to 39.5%, 39.5% to 40.5%, 40.5% to 41.5%, 41.5% to 45.0%, 45.0% to 46.5%, 46.5% to 50.0%, and 15.5%. Contains structural lipids in amounts (mol percent) of 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 nanoparticles contain structural lipids in an amount (mol percent) of about 50.0% to 55.0%, for example, about 50.0% to 51.5%, 51.5% to 53.5%, or 53.5% to 55.0%.

[0107] In some embodiments, the lipid nanoparticles contain PEG-lipids in amounts (mol percent) of about 0.5% to 5.0%, for example, about 0.5% to 1.0%, 1.0% to 1.5%, 1.5% to 2.0%, 2.0% to 2.5%, 2.5% to 3.0%, 3.0% to 3.5%, 3.5% to 4.0%, 4.0% to 4.5%, 4.5% to 5.0%, 1.5% to 1.6%, or 1.6% to 2.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids.

[0108] In some of the embodiments described above, the helper lipid is a phospholipid. Phospholipids are generally semi-synthetic and may be of natural origin or chemically modified. Examples of phospholipids include, but are not limited to, DSPC (distearoylphosphatidylcholine), DOPE (dioleoylphosphatidylethanolamine), DOPC (dioleoyllecithin), DOPS (dioleoylphosphatidylserine), DSPG (1,2-distearoyl-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), and lysophospholipids. Preferably, the helper lipid is one or more selected from the group consisting of DSPC, DOPE, DOPC, and DOPS. In some embodiments, the helper lipids are DSPC and / or DOPE.

[0109] In some embodiments, the structural lipid is a sterol that includes, but is not limited to, cholesterol, cholesterol esters, steroid hormones, steroid vitamins, bile acids, cholesterol, ergosterol, β-sitosterol, oxidized cholesterol derivatives, etc. Preferably, the structural lipid is at least one selected from cholesterol, cholesterol esters, steroid hormones, steroid vitamins, and bile acids. In some embodiments, the structural lipid is cholesterol, preferably high-purity cholesterol, in particular injection-grade high-purity cholesterol such as CHO-HP(AVT).

[0110] As used herein, the term PEG-lipid (polyethylene glycol-lipid) refers to 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, PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristoyl-sn-glycerol. Preferably, PEG has an average molecular weight of about 2,000 to 5,000, preferably about 2,000, for example, PEG2000-DMG.

[0111] In some of the embodiments described above, in lipid nanoparticles, the molar ratio of the aminolipid compound:helper lipid:structural lipid:PEG-lipid of the Disclosure is approximately 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 45.0:15.0:38. 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. The ratios are 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.

[0112] In the other embodiments described above, in lipid nanoparticles, the molar ratio of the aminolipid compound:helper lipid:structural lipid:PEG-lipid of the Disclosure is approximately 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 0.5:1.5, 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 The ratios are 0.5, 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.

[0113] In some embodiments, the lipid nanoparticles have the aminolipid compounds, helper lipids, structural lipids, and PEG-lipids of this disclosure in mole percent (%), as shown in numbers 1-12 and 13-24 of Table 2 below, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids: [Table 2] In some embodiments, the lipid nanoparticles have the aminolipid compounds, helper lipids, structural lipids, and PEG-lipids of the Disclosure in mole percent (%), as shown in numbers 25-42 of Table 3 below, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids: [Table 3] As described above, the lipid nanoparticles of this disclosure can be used as a vehicle for the active ingredient.

[0114] In some embodiments, the active ingredient includes a therapeutic agent and / or a prophylactic agent.

[0115] The terms “therapeutic” or “preventive” refer to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or preventive effect, and / or induces a desired biological and / or pharmacological effect.

[0116] “Effective dose” or “therapeutic effective dose” means an amount of the aminolipid compound of the Disclosure or lipid nanoparticles containing the aminolipid compound of the Disclosure that, when administered to a mammal (preferably a human), would produce a therapeutic effect in the mammal (preferably a human). The amount of lipid nanoparticles of the Disclosure constituting a “therapeutic effective dose” depends on the aminolipid compound, the condition and its severity, the mode of administration, and the age of the mammal being treated, but can be routinely determined by those skilled in the art in light of their knowledge and the Disclosure.

[0117] In some embodiments, the active ingredient is a pharmaceutically active ingredient that includes, but is not limited to, antineoplastic agents, antibiotics, immunomodulators, anti-inflammatory agents, drugs acting on the central nervous system, antigens or fragments thereof, proteins, peptides, polypeptides, polypeptides, vaccines and small molecules, and mixtures thereof. Preferably, the pharmaceutically active ingredient is a biologically active ingredient.

[0118] In the context of this disclosure, a biologically active component is a substance that, when introduced into a cell or host, has a biological effect, for example, by stimulating an immune or inflammatory response, by exerting enzymatic activity, or by complementing mutations. Examples of biologically active components include, but are not limited to, nucleic acids, antigens or their fragments, proteins, peptides, polypeptides, polypeptoids, antibodies, vaccines and small molecules, and mixtures thereof.

[0119] Lipid nanoparticles can be referred to as "lipid nanoparticle drugs" when they contain an active ingredient encapsulated within their internal aqueous space.

[0120] In the context of this disclosure, the term “cell” is a general term and includes individual cells, tissues, organs, insect cells, avian cells, fish cells, amphibian cells, mammalian cells, primary cells, serial cell lines, stem cells, and / or cultures of genetically modified cells (e.g., recombinant cells expressing heterologous polypeptides or proteins). Recombinant cells include, for example, cells expressing heterologous polypeptides or proteins (such as growth factors or blood factors).

[0121] In some preferred embodiments, the biologically active component is a nucleic acid.

[0122] In some of the embodiments described above, the lipid nanoparticles of this disclosure further comprise nucleic acids.

[0123] In some embodiments, the mass ratio of the aminolipid compound of the present disclosure to nucleic acid in lipid nanoparticles is about (5-30):1, for example, about (5-10):1, (10-15):1, (15-20):1, (20-25):1, or (25-30):1, preferably about 10:1.

[0124] In some embodiments, the nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides, and DNA.

[0125] In some embodiments, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), nuclear small RNA (snRNA), small hairpin RNA (shRNA), single guide RNA (sgRNA), Cas9 mRNA, or mixtures thereof.

[0126] In some embodiments, messenger RNA (mRNA) encodes the polypeptide and / or protein of interest. This includes any natural, non-natural, or otherwise modified polypeptide. In some embodiments, the polypeptide and / or protein encoded by mRNA may have therapeutic and / or preventive effects when expressed in cells.

[0127] In some embodiments, the RNA is an siRNA that can selectively reduce the expression of a gene of interest, or downregulate its expression. For example, siRNA can be selected so that when lipid nanoparticles containing the siRNA are administered to a target that needs it, genes associated with a particular disease, disorder, or condition are silenced. The siRNA may contain a sequence complementary to the mRNA sequence encoding the gene or protein of interest. In some embodiments, the siRNA may be an immunomodulatory siRNA.

[0128] 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 mRNA translation of cellular genes.

[0129] In some embodiments, the RNA is shRNA or a vector or plasmid encoding it. The shRNA may be produced within the target cell after a suitable construct has been delivered to the nucleus. Constructs and mechanisms associated with shRNA are well known in the relevant art.

[0130] In some embodiments, DNA is a plasmid.

[0131] In some embodiments, lipid nanoparticles are used to transfer nucleic acids. In some embodiments, lipid nanoparticles may be used, for example, in gene therapy, gene vaccination, protein replacement therapy, antisense therapy, or interfering RNA therapy.

[0132] composition In another aspect, the disclosure provides a composition comprising the above-mentioned lipid nanoparticles and a pharmaceutically acceptable carrier, diluent, or excipient.

[0133] In some embodiments, the composition further comprises a buffer solution. In some such embodiments, the buffer solution is selected from phosphate buffer and Tris buffer, preferably phosphate buffer. In some embodiments, the buffer solution has a concentration of about 5 mmol / L to about 30 mmol / L, preferably about 10 mmol / L. In some embodiments, the buffer solution has a pH of about 6 to 8, preferably about 7 to 8, more preferably about 7 to 7.5.

[0134] 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 cryoprotectant has a concentration of about 50 mg / ml to 100 mg / ml.

[0135] In some embodiments 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 cryoprotectant has a concentration of about 50 mg / ml to 100 mg / ml.

[0136] In some embodiments, the composition is a pharmaceutical composition.

[0137] Purpose The lipid nanoparticles of this disclosure have excellent properties for encapsulating biologically active ingredients. Lipid nanoparticles containing biologically active ingredients can be used to transfer any of a variety of therapeutic agents into cells. This disclosure includes the use of the above-mentioned lipid nanoparticles for transferring biologically active ingredients into cells. This disclosure also provides a method for transferring biologically active ingredients into cells, which includes contacting the cells with the lipid nanoparticles of this disclosure containing the biologically active ingredient.

[0138] The lipid nanoparticles of this disclosure can be used to transfect multicellular tissues or organs. Accordingly, this disclosure also provides a method for transfecting cells, multicellular tissues or organs, comprising contacting the cells, tissues or organs with the lipid nanoparticles of this disclosure, which contain nucleic acids. This offers the possibility of novel therapeutic treatments for subjects.

[0139] In some embodiments, the cells are mammalian cells, and more preferably, the mammalian cells are located within a mammal.

[0140] In some embodiments, the tissue or organ is selected from the group consisting of the spleen, liver, kidney, lung, femur, eye tissue, vascular endothelium in blood vessels, lymph, and tumor tissue.

[0141] As used herein, the subject may be any mammal selected from the group consisting of mice, rats, pigs, cats, dogs, horses, goats, cattle, and monkeys, among others. In some preferred embodiments, the subject is a human.

[0142] In another aspect, the Disclosure provides a method for producing a target polypeptide and / or protein in mammalian cells, comprising contacting cells with lipid nanoparticles containing mRNA encoding the target polypeptide and / or protein, wherein the mRNA can be taken up by the cells and translated to produce the target polypeptide and / or protein.

[0143] In yet another embodiment, the Disclosure provides the use of the aminolipid compounds, lipid nanoparticles, or compositions of the Disclosure in the manufacture of a pharmaceutical product. In yet another embodiment, the Disclosure provides lipid nanoparticles or compositions of the Disclosure for use as a pharmaceutical product. In yet another embodiment, the Disclosure provides a pharmaceutical product comprising the lipid nanoparticles or compositions of the Disclosure. In some embodiments, the pharmaceutical product 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, renovascular diseases, and metabolic diseases. In some embodiments, the pharmaceutical product is used, for example, for gene therapy, gene vaccination, protein replacement therapy, antisense therapy, or interfering RNA therapy. Preferably, gene therapy is for use in the treatment of cancer and genetic diseases. In some embodiments, cancer is selected from one or more of lung cancer, gastric cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, hematological cancer, or prostate cancer. In some embodiments, the genetic disorder is one or more selected from hemophilia, thalassemia, and Gaucher disease. In some embodiments, the gene vaccine is preferably for vaccination to treat cancer, allergies, toxic diseases, and pathogenic infections. In some embodiments, the pathogen is selected from one or more viruses, bacteria, or fungi.

[0144] In yet another aspect, the disclosure provides a method for treating a disease or disorder in a mammal requiring treatment, comprising administering a therapeutically effective amount of the above-mentioned lipid nanoparticles to the mammal.

[0145] Preferably, the disease or disorder is selected from the group consisting of rare diseases, infectious diseases, cancer, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renovascular diseases, and metabolic diseases.

[0146] In yet another aspect, the present disclosure provides for the use of the amino lipid compounds, lipid nanoparticles or compositions of the present disclosure in the manufacture of a medicament for nucleic acid transfection. In yet another aspect, the present disclosure provides the lipid nanoparticles or compositions of the present disclosure for use as a medicament for nucleic acid transfection. In yet another aspect, the present disclosure provides a medicament comprising the lipid nanoparticles or compositions of the present disclosure for nucleic acid transfection. In some embodiments, the nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides and DNA. In some embodiments, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA) and small nuclear RNA (snRNA). In some embodiments, the DNA is a plasmid.

[0147] Preparation method In another aspect, the present disclosure also provides a general synthetic process for preparing the amino lipid compounds of formula (II) of the present disclosure as follows:

Chemical formula

[0148] Specifically, the method comprises Step one: subjecting a compound of formula DTN and a compound of formula di-N to a reductive amination reaction to obtain a compound of formula F; and Step two: reacting the compound of formula F with a compound of formula G to obtain the amino lipid compound of formula (II).

[0149] 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 a hydrogen atmosphere in the presence of a catalyst (e.g., palladium-carbon). In some embodiments, the reductive amination reaction is carried out under high pressure (e.g., about 2-8 or about 4-6 MPa) and high temperature (e.g., about 50-90 or about 60-80°C).

[0150] In some embodiments, the reaction in step 2 is carried out in the presence of a base (e.g., pyridine or DMAP).

[0151] In yet another aspect, the Disclosure also provides a general synthetic process for preparing aminolipid compounds of formula (III) of the Disclosure, as follows: [ka] (In the formulas, A3, A4, A6, A7, R1, R2, R3, R4, R5, Z7, and Z8 have the same meanings as defined above for compounds of formula (I) or (III).)

[0152] Specifically, this method is Step 1: The compound of formula DTN and the compound of formula di-N are subjected to a reductive amination reaction to obtain the compound of formula F. Step 2: The step of reacting a compound of formula F with a compound of formula H to obtain an aminolipid compound of formula (III).

[0153] 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 a hydrogen atmosphere in the presence of a catalyst (e.g., palladium-carbon). In some embodiments, the reductive amination reaction is carried out under high pressure (e.g., about 2-8 or about 4-6 MPa) and high temperature (e.g., about 50-90 or about 60-80°C).

[0154] In yet another embodiment, the lipid nanoparticles or compositions of the present disclosure may be prepared according to methods known in the art. For example, the method may include the following steps:

[0155] (1) Formulation: A step of forming an organic phase comprising an aminolipid compound of the present disclosure and optionally a helper lipid, structural lipid and / or PEG-lipid, by forming a suitable aqueous phase. (2) Sealing: A step of mixing a suitable amount of aqueous phase with the organic phase, (3) Dialysis: Optionally, a step of dialysis of the mixture from step (2), and (4) Sterilization: Optionally, sterilize the product from step (3) using a sterilization filter, such as a 0.22 μm microporous membrane.

[0156] In yet another embodiment, lipid nanoparticles or compositions of the present disclosure, comprising nucleic acids, particularly mRNA, may be prepared by a method comprising the following steps:

[0157] (1) Formulation: A step of formulation an aqueous phase containing nucleic acids, and an organic phase (e.g., an ethanol phase) containing the aminolipid compound of the Disclosure and optionally helper lipids, structural lipids and / or PEG-lipids, (2) Sealing: A step of mixing a suitable amount of aqueous phase with the organic phase, (3) Dialysis: Optionally, a step of dialysis of the mixture from step (2), (4) Sterilization: Optionally, sterilize the product from step (3) using a sterilization filter, such as a 0.22 μm microporous membrane.

[0158] This disclosure also includes the following embodiments.

[0159] Embodiment 1. Aminolipid compound having the structure of formula (I): [ka] (In the formula, 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)-, -S-S-, or a bond, Z3 is -C(=O)- or a bond, Z4 is -O- or -CH(OH)-, A1, A2, A3, A4, A5, A6, and A7 are each independently a C1-C 12 hydrocarbylene, cyclo-hydrocarbyl, phenyl, heterocycle, or a bond, R1 and R2 are each independently H or a C1-C 18 hydrocarbyl or cyclo-hydrocarbyl, phenyl, heterocycle, or R1 and R2 together with the nitrogen atom to which they are attached form a 4- to 7-membered heterocycle, R3 is H or a C1-C 18 hydrocarbyl or cyclo-hydrocarbyl, phenyl, heterocycle, R4 and R5 are each independently a C1-C 24 hydrocarbyl or cyclo-hydrocarbyl, phenyl, heterocycle, preferably, A5, Z5, and Z6 are bonds) or a pharmaceutically acceptable salt or stereoisomer thereof.

[0160] Embodiment 2. The amino lipid compound according to Embodiment 1, wherein Z7 and Z8 are each independently -(C=O)O-, -O(C=O)-, or a bond.

[0161] Embodiment 3. The amino lipid compound according to Embodiment 1, wherein one of Z7 or Z8 is -(C=O)O-, -O(C=O)-, or a bond.

[0162] Embodiment 4. The amino lipid compound according to Embodiment 1, wherein each of Z7 and Z8 is -(C=O)O-.

[0163] Embodiment 5. The aminolipid compound according to Embodiment 1, wherein one of Z7 and Z8 is a bond.

[0164] Embodiment 6. The aminolipid compound according to Embodiment 1, wherein Z7 and Z8 are each bonds.

[0165] Embodiment 7. R4 and R5 are independently C1-C 20 The aminolipid compound described in Embodiment 1, which is hydrocarbyl.

[0166] Embodiment 8. R4 and R5 are independently C1-C 20 The aminolipid compound described in Embodiment 1, which is alkyl.

[0167] Embodiment 9. R4 and R5 are independently connected to branch C3-C 20 The aminolipid compound described in Embodiment 1, which is alkyl.

[0168] Embodiment 10. R4 and R5 are independently, [ka] The aminolipid compound described in Embodiment 1.

[0169] Embodiment 11. Aminolipid compound having the structure of formula (II): [ka] (In the formula, Z7 and Z8 are 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 Hydrocarbylene, cyclohydrocarbyl, phenyl, heterocyclic or bonded, R1 and R2 are independently H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, or R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocyclic ring. R3 is H or C1-C 18 Hydrocarbyl, or cyclohydrocarbyl, phenyl, heterocyclic, R4 and R5 are independently C1-C 24 Hydrocarbyl or cyclohydrocarbyl, phenyl (which is heterocyclic), or its pharmaceutically acceptable salts or stereoisomers.

[0170] Embodiment 12. The aminolipid compound according to Embodiment 11, wherein Z7 and Z8 are independently -(C=O)O- or -O(C=O)-.

[0171] Embodiment 13. A3, A4, A6, and A7 are each independently C1-C 12 The aminolipid compound according to Embodiment 11, which is an alkylene.

[0172] Embodiment 14. R1 and R2 are independently C1-C 18 The aminolipid compound described in Embodiment 11, which is alkyl.

[0173] Embodiment 15. The aminolipid compound according to Embodiment 14, wherein R1 and R2 are each independently a C1-C6 alkyl group.

[0174] Embodiment 16. The aminolipid compound according to Embodiment 14, wherein R1 and R2 are each independently methyl, ethyl, n-propyl, or isopropyl.

[0175] Embodiment 17. The aminolipid compound according to Embodiment 11, wherein R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocycle.

[0176] Embodiment 18. The aminolipid compound according to Embodiment 17, wherein R1 and R2, together with the nitrogen atom to which they are bonded, form a 5-6 membered heterocycle.

[0177] Embodiment 19.R3 is C1-C 18 The aminolipid compound described in Embodiment 11, which is alkyl.

[0178] Embodiment 20. R4 and R5 are independently C1-C 20 The aminolipid compound described in Embodiment 11, which is hydrocarbyl.

[0179] Embodiment 21. R4 and R5 are independently C1-C 20 The aminolipid compound described in Embodiment 11, which is alkyl.

[0180] Embodiment 22. R4 and R5 are independently connected to branch C3-C 18 The aminolipid compound described in Embodiment 11, which is alkyl.

[0181] Embodiment 23. R4 and R5 are independently, [ka] The aminolipid compound described in Embodiment 11.

[0182] Embodiment 24. Aminolipid compound having the structure of formula (III): [ka] (In the formula, Z7 and Z8 are 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 12Hydrocarbylene, cyclohydrocarbyl, phenyl, heterocyclic or bonded, R1 and R2 are independently H or C1-C 18 Hydrocarbyl or cyclohydrocarbyl, phenyl, heterocyclic, or R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocyclic ring. R3 is H or C1-C 18 Hydrocarbyl, or cyclohydrocarbyl, phenyl, heterocyclic, R4 and R5 are independently C1-C 24 Hydrocarbyl or cyclohydrocarbyl, phenyl (which is heterocyclic), or its pharmaceutically acceptable salts or stereoisomers.

[0183] Embodiment 25. The aminolipid compound according to Embodiment 24, wherein Z7 and Z8 are independently -(C=O)O- or -O(C=O)-.

[0184] Embodiment 26. A3, A4, A6, and A7 are each independently C1-C 12 The aminolipid compound according to Embodiment 24, which is alkylene.

[0185] Embodiment 27. R1 and R2 are independently C1-C 18 The aminolipid compound according to Embodiment 24, which is alkyl.

[0186] Embodiment 28. The aminolipid compound according to Embodiment 27, wherein R1 and R2 are each independently a C1-C6 alkyl group.

[0187] Embodiment 29. The aminolipid compound according to Embodiment 28, wherein R1 and R2 are each independently methyl, ethyl, n-propyl, or isopropyl.

[0188] Embodiment 30. The aminolipid compound according to Embodiment 24, wherein R1 and R2, together with the nitrogen atom to which they are bonded, form a 4- to 7-membered heterocycle.

[0189] Embodiment 31. The aminolipid compound according to Embodiment 24, wherein R1 and R2, together with the nitrogen atom to which they are bonded, form a 5-6 membered heterocycle.

[0190] Embodiment 32.R3 is C1-C 18 The aminolipid compound according to Embodiment 24, which is alkyl.

[0191] Embodiment 33. R4 and R5 are independently C1-C 20 The aminolipid compound according to Embodiment 24, which is hydrocarbyl.

[0192] Embodiment 34. R4 and R5 are independently C1-C 20 The aminolipid compound according to Embodiment 24, which is alkyl.

[0193] Embodiment 35. R4 and R5 are independently connected to branch C3-C 18 The aminolipid compound according to Embodiment 24, which is alkyl.

[0194] Embodiment 36. R4 and R5 are independently, [ka] The aminolipid compound described in Embodiment 24.

[0195] Embodiment 37. Lipid nanoparticles comprising an aminolipid compound according to any one of Embodiments 1 to 36 and optionally a pharmaceutically acceptable carrier, diluent, or excipient.

[0196] Embodiment 38. Further comprising one or more of helper lipids, structural lipids, and PEG-lipids, Furthermore, the lipid nanoparticles according to Embodiment 37 include a helper lipid, a structural lipid, and a PEG-lipid.

[0197] Embodiment 39. Lipid nanoparticles according to Embodiment 38, wherein the lipid nanoparticles contain the aminolipid compound described in any one of Embodiments 1 to 36 in an amount (mol percent) of about 25.0% to 75.0%, for example, about 25.0% to 28.0%, 28.0% to 32.0%, 32.0% to 35.0%, 35.0% to 40.0%, 40.0% to 42.0%, 42.0% to 45.0%, 45.0% to 48.0%, 48.0% to 55.0%, 55.0% to 65.0%, or 65.0% to 75.0%, based on the total amount of the aminolipid compound, helper lipid, structural lipid and PEG-lipid described in any one of Embodiments 1 to 36.

[0198] Embodiment 40. Lipid nanoparticles according to Embodiment 38 or 39, wherein the lipid nanoparticles contain helper lipids in an amount (mol percent) 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%, or 42.0% to 45.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0199] Embodiment 41. Lipid nanoparticles according to any one of Embodiments 38 to 40, wherein the lipid nanoparticles contain structural lipids in an amount (mol percent) of about 0.0% to 50.0%, for example, about 0.0% to 10.0%, 10.0% to 15.5%, 15.5% to 22.5%, 22.5% to 35.0%, 35.0% to 36.5%, 36.5% to 39.5%, 39.5% to 40.5%, 40.5% to 41.5%, 41.5% to 45.0%, 45.0% to 46.5%, or 46.5% to 50.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0200] Embodiment 42. Lipid nanoparticles according to any one of Embodiments 38 to 41, wherein the lipid nanoparticles contain PEG-lipids in an amount (mol percent) of about 0.5% to 5.0%, for example, about 0.5% to 1.0%, 1.0% to 1.5%, 1.5% to 2.0%, 2.0% to 2.5%, 2.5% to 3.0%, 3.0% to 3.5%, 3.5% to 4.0%, 4.0% to 4.5%, or 4.5% to 5.0% based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0201] Embodiment 43. Lipid nanoparticles according to any one of Embodiments 38 to 42, wherein the helper lipid is one or more selected from the group consisting of DSPC, DOPE, DOPC, DOPS, DSPG, DPPG, DPPC, DGTS, and lysophospholipids, preferably one or more selected from the group consisting of DSPC, DOPE, DOPC, and DOPS, more preferably DSPC and / or DOPE.

[0202] Embodiment 44. A lipid nanoparticle according to any one of Embodiments 38 to 43, wherein the structural lipid is at least one selected from the group consisting of cholesterol, cholesterol esters, steroid hormones, steroid vitamins, bile acids, cholesterol, ergosterol, β-sitosterol, and oxidized cholesterol derivatives, preferably at least one selected from the group consisting of 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.

[0203] Embodiment 45. The PEG-lipid is selected from PEG-DMG and PEG-DSPE, preferably PEG-DMG. Preferably, PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristoyl-sn-glycerol. Preferably, the lipid nanoparticles according to any one of embodiments 38 to 44, wherein the PEG has an average molecular weight of about 2,000 to 5,000, preferably about 2,000.

[0204] Embodiment 46. The molar ratio of the aminolipid compound:helper lipid:structural lipid:PEG-lipid described in any one of Embodiments 1 to 36 is approximately 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. Lipid nanoparticles according to any one of embodiments 38 to 45, which are 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.

[0205] Embodiment 47. The lipid nanoparticle according to Embodiment 46, wherein the helper lipid is DOPE and the structural lipid is CHO-HP.

[0206] Embodiment 48. Lipid nanoparticles according to any one of Embodiments 38 to 45, wherein the molar ratio of aminolipid compound:helper lipid:structural lipid:PEG-lipid according to any one of Embodiments 1 to 36 is approximately 48.0:10.0:40.5:1.5.

[0207] Embodiment 49. Lipid nanoparticles according to Embodiment 48, wherein the helper lipid is DSPC and the structural lipid is CHO-HP.

[0208] Embodiment 50. Lipid nanoparticles according to any one of Embodiments 37 to 49, further comprising nucleic acids.

[0209] Embodiment 51. Lipid nanoparticles according to Embodiment 50, wherein the mass ratio of the aminolipid compound to nucleic acid according to any one of Embodiments 1 to 36 is about (5 to 30):1, for example about (5 to 10):1, (10 to 15):1, (15 to 20):1, (20 to 25):1 or (25 to 30):1, preferably about 10:1.

[0210] Embodiment 52. The nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides, and DNA. Preferably, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), and nuclear small RNA (snRNA). Preferably, the lipid nanoparticles according to embodiment 50 or 51, wherein the DNA is a plasmid.

[0211] Embodiment 53. A composition comprising an aminolipid compound according to any one of Embodiments 1 to 36 and a pharmaceutically acceptable carrier, diluent, or excipient.

[0212] Embodiment 54. Further comprising one or more of helper lipids, structural lipids, and PEG-lipids, Furthermore, the composition according to Embodiment 53, wherein the composition further comprises a helper lipid, a structural lipid, and a PEG-lipid.

[0213] Embodiment 55. The composition according to Embodiment 54, wherein the composition contains an aminolipid compound described in any one of Embodiments 1 to 36 in an amount (mol percent) of about 25.0% to 75.0%, for example, about 25.0% to 28.0%, 28.0% to 32.0%, 32.0% to 35.0%, 35.0% to 40.0%, 40.0% to 42.0%, 42.0% to 45.0%, 45.0% to 48.0%, 48.0% to 55.0%, 55.0% to 65.0%, or 65.0% to 75.0%, based on the total amount of the aminolipid compound, helper lipid, structural lipid and PEG-lipid described in any one of Embodiments 1 to 36.

[0214] Embodiment 56. The composition according to Embodiment 54 or 55, wherein the composition contains helper lipids in an amount (mol percent) 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%, or 42.0% to 45.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0215] Embodiment 57. The composition according to any one of Embodiments 54 to 56, wherein the composition contains structural lipids in an amount (mol percent) of about 0.0% to 50.0%, for example, about 0.0% to 10.0%, 10.0% to 15.5%, 15.5% to 22.5%, 22.5% to 35.0%, 35.0% to 36.5%, 36.5% to 39.5%, 39.5% to 40.5%, 40.5% to 41.5%, 41.5% to 45.0%, 45.0% to 46.5%, or 46.5% to 50.0%, based on the total amount of amino lipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0216] Embodiment 58. The composition according to any one of Embodiments 54 to 57, wherein the composition contains PEG-lipids in an amount (mol percent) of about 0.5% to 5.0%, for example, about 0.5% to 1.0%, 1.0% to 1.5%, 1.5% to 2.0%, 2.0% to 2.5%, 2.5% to 3.0%, 3.0% to 3.5%, 3.5% to 4.0%, 4.0% to 4.5%, or 4.5% to 5.0%, based on the total amount of aminolipid compounds, helper lipids, structural lipids, and PEG-lipids described in any one of Embodiments 1 to 36.

[0217] Embodiment 59. The composition according to any one of Embodiments 54 to 58, wherein the helper lipid is one or more selected from the group consisting of DSPC, DOPE, DOPC, DOPS, DSPG, DPPG, DPPC, DGTS, and lysophospholipids, preferably one or more selected from the group consisting of DSPC, DOPE, DOPC, and DOPS, more preferably DSPC and / or DOPE.

[0218] Embodiment 60. The composition according to any one of Embodiments 54 to 58, wherein the structural lipid is at least one selected from the group consisting of cholesterol, cholesterol esters, steroid hormones, steroid vitamins, bile acids, cholesterol, ergosterol, β-sitosterol, and oxidized cholesterol derivatives, preferably at least one selected from the group consisting of 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.

[0219] Embodiment 61. The PEG-lipid is selected from PEG-DMG and PEG-DSPE, preferably PEG-DMG. Preferably, PEG-DMG is a polyethylene glycol (PEG) derivative of 1,2-dimyristoyl-sn-glycerol. Preferably, the composition according to any one of embodiments 54 to 58, wherein the PEG has an average molecular weight of about 2,000 to 5,000, preferably about 2,000.

[0220] Embodiment 62. The molar ratio of aminolipid compound:helper lipid:structural lipid:PEG-lipid described in any one of Embodiments 1 to 36 is approximately 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. The composition according to any one of Embodiments 54 to 61, wherein the ratio is 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.

[0221] Embodiment 63. The composition according to Embodiment 62, wherein the helper lipid is DOPE and the structural lipid is CHO-HP.

[0222] Embodiment 64. The composition according to any one of Embodiments 54 to 61, wherein the molar ratio of the aminolipid compound, helper lipid, structural lipid, and PEG-lipid according to any one of Embodiments 1 to 36 is approximately 48.0:10.0:40.5:1.5.

[0223] Embodiment 65. The composition according to Embodiment 64, wherein the helper lipid is DSPC and the structural lipid is CHO-HP.

[0224] Embodiment 66. The composition according to any one of Embodiments 53 to 65, further comprising nucleic acid.

[0225] Embodiment 67. The composition according to Embodiment 66, wherein the mass ratio of the aminolipid compound to nucleic acid according to any one of Embodiments 1 to 36 is about (5 to 30):1, for example about (5 to 10):1, (10 to 15):1, (15 to 20):1, (20 to 25):1 or (25 to 30):1, preferably about 10:1.

[0226] Embodiment 68. The nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides, and DNA. Preferably, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), and nuclear small RNA (snRNA). Preferably, the composition according to embodiment 66 or 67, wherein the DNA is a plasmid.

[0227] Embodiment 69. Further comprising a buffer solution, Preferably, the buffer is selected from phosphate buffer and Tris buffer, preferably phosphate buffer, and / or Preferably, the buffer solution has a concentration of about 5 mmol / L to 30 mmol / L, preferably about 10 mmol / L, and / or Preferably, the buffer solution has a pH of about 6 to 8, preferably about 7 to 8, and more preferably about 7 to 7.5, according to any one of embodiments 53 to 68.

[0228] Embodiment 70. Further comprising a cryoprotectant, Preferably, the cryoprotectant is selected from sucrose and trehalose, preferably sucrose, and / or Preferably, the composition according to any one of embodiments 53 to 68, wherein the cryoprotectant has a concentration of about 50 mg / ml to 100 mg / ml.

[0229] Embodiment 71. Use of an aminolipid compound according to any one of Embodiments 1 to 36 in the production of a vehicle for an active ingredient.

[0230] Embodiment 72. The vehicle is a lipid nanoparticle described in any one of Embodiments 37 to 52, and / or The active ingredient is a pharmaceutical active ingredient, preferably a nucleic acid, and / or Preferably, the nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides, and DNA. Preferably, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), and nuclear small RNA (snRNA). Preferably, the use according to Embodiment 71, wherein the DNA is a plasmid.

[0231] Embodiment 73. Use of an aminolipid compound according to any one of Embodiments 1 to 36, lipid nanoparticles according to any one of Embodiments 37 to 52, or a composition according to any one of Embodiments 53 to 70 in the manufacture of a pharmaceutical product.

[0232] Embodiment 74. The use according to Embodiment 73, wherein the pharmaceutical is for use in gene therapy, gene vaccination, protein replacement therapy, antisense therapy, or interfering RNA therapy.

[0233] Embodiment 75. The use of the gene therapy described in Embodiment 74, wherein the gene therapy is for use in the treatment of cancer and genetic diseases.

[0234] Embodiment 76. The use according to Embodiment 75, wherein the cancer is one or more selected from the group consisting of lung cancer, stomach cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, hematological cancer, or prostate cancer, and the hereditary disease is one or more selected from the group consisting of hemophilia, thalassemia, and Gaucher disease.

[0235] Embodiment 77. The use of the gene vaccine according to Embodiment 75, wherein the gene vaccine is for use in the treatment of cancer, allergies, toxicology, and pathogen infections.

[0236] Embodiment 78. The use according to Embodiment 77, wherein the pathogen is one or more selected from the group consisting of viruses, bacteria, or fungi.

[0237] Embodiment 79. Use of an aminolipid compound according to any one of Embodiments 1 to 36, a lipid nanoparticle according to any one of Embodiments 37 to 52, or a composition according to any one of Embodiments 53 to 70 in the manufacture of a pharmaceutical product for nucleic acid transfer.

[0238] Embodiment 80. The nucleic acid is selected from the group consisting of RNA, antisense oligonucleotides, and DNA. Preferably, the RNA is selected from the group consisting of messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), and nuclear small RNA (snRNA). Preferably, the use according to Embodiment 79, wherein the DNA is a plasmid.

[0239] Beneficial effects The aminolipid compounds of this disclosure can form vehicles such as lipid nanoparticles that have excellent properties for encapsulating biologically active ingredients, and can be used to deliver biologically active ingredients, particularly water-insoluble drugs or readily degradable or reduced active ingredients (such as nucleic acids), and to improve their bioavailability and efficacy, or immunological activity, or transfection efficiency (in the case of nucleic acids), or safety, or preference for specific organs or tissues. In particular, lipid nanoparticles containing the aminolipid compounds of this disclosure exhibit clear delivery preference to the spleen.

[0240] Lipid nanoparticles containing the aminolipid compounds of this disclosure exhibit clear delivery preference to immune cells in different tissues and organs, including the spleen, lymph, peripheral blood, bone marrow, lungs, and liver, and immune cells including neutrophils, NK cells-NK T cells, macrophages-monocytes, dendritic cells, B cells, T cells, and CD4 T cells.

[0241] Figures 1A to 1F show the delivery efficiency of lipid nanoparticles containing different aminolipid compounds to immune cell populations in different tissues and organs. [Brief explanation of the drawing]

[0242] [Figure 1A] This is a diagram of neutrophils. [Figure 1B] NK cell-NK T cell diagram. [Figure 1C] This is a diagram showing macrophages and monocytes. [Figure 1D] This is a diagram showing dendritic cells. [Figure 1E] This figure shows B cells. [Figure 1F] This figure shows CD4 T cells.

[0243] To further clarify the purpose, technical solutions, and benefits of this disclosure, the disclosure is described below with reference to specific examples. The following examples are merely illustrative and not intended to limit the scope of this disclosure.

[0244] Examples The following examples are provided for illustrative purposes only and not for limitation.

[0245] In the examples, experimental methods for which specific conditions are not specified are typically conventional conditions or conditions as recommended by the raw material or product manufacturer, and reagents of unspecified origin are generally conventional reagents that are commercially available.

[0246] The abbreviations used in the examples have the following meanings:

[0247] Pd / C Palladium / Carbon, EA ethyl acetate, DCM dichloromethane, MPa megapascals, DMF N,N-dimethylformamide, EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, DMAP 4-dimethylaminopyridine, TEMPO 2,2,6,6-tetramethylpiperidinooxy NaDCC (Sodium Dichloroisocyanurate) h time, min (minutes).

[0248] Example 1: Synthesis of aminolipid compound 125 (1) Synthesis of compound 1522F-1 [ka] 10.0 g of DTN-T (14.1 mmol), 2.88 g of N,N-dimethylaminopropylamine, 0.5 g of Pd / C (10%), and 80 ml of anhydrous ethanol were added to a 200 ml autoclave, sealed, and purged three times with nitrogen, then three times with hydrogen, then filled with hydrogen to a pressure of 4 MPa, and then heated to 70°C. After reacting for 48 hours, suction filtration was performed, the filtrate was washed with anhydrous ethanol, and the filtrate was concentrated to obtain 11 g of crude product. The crude product was purified by silica gel column chromatography and eluted with EA:methanol = 9:1 to obtain 9 g of 1522F-1 in 80% yield.

[0249] (2) Synthesis of compound 125-G [ka] 8.67 g of sodium hydride and 195 ml of DMF were added to a 500 mL two-necked flask, and the apparatus was cooled to 0°C under nitrogen protection. Then, 15.0 g of 1,3-propanediol (197.1 mmol) was slowly added. After reacting for 30 minutes, the mixture was heated to room temperature and continued to react for 1 hour, then cooled to 0°C. 35.7 g of 1-bromopentane was slowly added dropwise to the reaction mixture. After reacting for 30 minutes, the mixture was heated to room temperature and continued to react for 15 hours. 100 mL of water was added, and the mixture was extracted with 100 mL of EA, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was subjected to silica gel column chromatography, and eluted with EA:n-hexane = 1:8 to obtain 17.10 g of 125G-1 in yield 59.3%.

[0250] 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 were added to a 1000 ml round-bottom flask. 38.34 g of NaDCC aqueous solution was added while stirring, and the mixture was reacted at room temperature for 15 hours with stirring, then filtered. The organic phase was separated from the filtrate, and the aqueous phase was extracted twice with 100 ml of DCM. The organic phases were combined and concentrated under reduced pressure. The concentrate was subjected to silica gel column chromatography, and eluted with EA:n-hexane = 1:8 to obtain 11 g of 125-G in 64.7% yield. 1 H NMR in(CDCl3)δ 3.71-3.69(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).

[0251] (3) Synthesis of aminolipid compound 125 [ka] 0.47 g of EDCI, 0.02 g of DMAP, and 30 ml of DCM were sequentially added to a 100 ml round-bottom flask and stirred for 10 minutes. Then, a solution of 1522F-1 (1.3 g, 1.64 mmol) in 3 ml of DCM, a solution of 125-G (0.34 g) in 3 ml of DCM, and a solution of pyridine (0.2 g) in 3 ml of DCM were sequentially added dropwise. After stirring at room temperature for 15 hours, the mixture was adjusted to pH 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 on anhydrous sodium sulfate and concentrated under reduced pressure to obtain 1.5 g of crude product. The crude product was subjected to silica gel column chromatography and eluted with EA:n-hexane = 1.5:1 to obtain aminolipid compound 125 (1.1 g) with a purity of 95.88% and a yield of 72%. 1H NMR(CDCl3)δ 4.86(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). LC-MS(ESI):(M+H)936.4.

[0252] Example 2: Synthesis of aminolipid compound 120 [ka] Following a general synthesis process, aminolipid compound 120 (0.56 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.31 g of 120-G with a purity of 90.85% and a yield of 37%. 1 H NMR(CDCl3)δ 4.86-4.84(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). LC-MS(ESI):(M+H)922.4.

[0253] Example 3: Synthesis of aminolipid compound 130 [ka] Following a general synthesis process, aminolipid compound 130 (1.05 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.37 g of 130-G with a purity of 92.04% and a yield of 67%. 1H NMR(CDCl3)δ 4.87-4.85(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). LC-MS(ESI):(M+H)950.4.

[0254] Example 4: Synthesis of aminolipid compound 144 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.87-4.85(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). LC-MS(ESI):(M+H)894.3.

[0255] Example 5: Synthesis of aminolipid compound 149 [ka] Following a general synthesis process, aminolipid 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%. 1H NMR(CDCl3)δ 4.87-4.85(m,2H),3.75-3.72(m,2H),3.63-3.61(m,1H),3.45-3.41(m,2H),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). LC-MS(ESI):(M+H)908.3.

[0256] Example 6: Synthesis of aminolipid compound 154 [ka] Following a general synthesis process, aminolipid compound 154 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.34 g of 154-G as a colorless oil with a purity of 93.80% and a yield of 66%. 1 H NMR(CDCl3)δ 4.87-4.85(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). LC-MS(ESI):(M+H)922.3.

[0257] Example 7: Synthesis of aminolipid compound 166 [ka] Following a general synthesis process, aminolipid 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%. 1H NMR(CDCl3)δ 4.87-4.84(m,2H),4.10(d,J=20 Hz,1H),3.50-3.47(m,3H),3.09-3.07(m,2H),2.30-2.25(m,6H),2.22(d,J=3.7 Hz,6H),1.61-1.36(m,24H),1.29-1.25(m,52H),0.92-0.85(m,15H). LC-MS(ESI):(M+H)922.3.

[0258] Example 8: Synthesis of aminolipid compound 172 [ka] Following a general synthesis process, aminolipid compound 172 (0.43 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.37 g of 125-G as a colorless oil with a purity of 97.49% and a yield of 28%. 1 H NMR(CDCl3)δ 4.87-4.85(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). LC-MS(ESI):(M+H)950.4.

[0259] Example 9: Synthesis of aminolipid compound 176 [ka] Following a general synthesis process, aminolipid 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%. 1H NMR(CDCl3)δ 4.87-4.85(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). LC-MS(ESI):(M+H)950.4.

[0260] Example 10: Synthesis of aminolipid compound 119 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.86-4.85(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). LC-MS(ESI):(M+H)908.4.

[0261] Example 11: Synthesis of aminolipid compound 121 [ka] Following a general synthesis process, aminolipid compound 121 (0.88 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.34 g of 121-G as a colorless oil with a purity of 92.94% and a yield of 57%. 1H NMR(CDCl3)δ 4.86-4.85(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). LC-MS(ESI):(M+H)936.5.

[0262] Example 12: Synthesis of aminolipid compound 124 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.86-4.85(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). LC-MS(ESI):(M+H)922.3.

[0263] Example 13: Synthesis of aminolipid compound 129 [ka] Following a general synthesis process, aminolipid compound 129 (0.79 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.34 g of 154-G as a colorless oil with a purity of 92.29% and a yield of 51%. 1H NMR(CDCl3)δ 4.87-4.85(m,2H),3.65(p,J=7.0 Hz,1H),3.44(q,J=6.2 Hz,2H),3.39(t,J=6.6 Hz,2H),3.13-3.10(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). LC-MS(ESI):(M+H)936.4.

[0264] Example 14: Synthesis of aminolipid compound 131 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.88-4.84(m,2H),3.65(p,J=7.0 Hz,1H),3.44(q,J=6.2 Hz,2H),3.38(t,J=6.7 Hz,2H),3.13-3.10(m,2H),2.43-2.37(m,2H),2.31-2.22(m,12H),1.95-1.88(m,2 H),1.74-1.67(m,2H),1.61-1.44(m,18H),1.32-1.26(m,58H),0.89-0.86(m,15H). LC-MS(ESI):(M+H)964.4.

[0265] Example 15: Synthesis of aminolipid compound 132 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.88-4.84(m,2H),3.65(p,J=7.0 Hz,1H),3.44(q,J=6.2 Hz,2H),3.38(t,J=6.6 Hz,2H),3.13-3.10(m,2H),2.43-2.37(m,2H),2.30-2.22(m,12H),1.95-1.88(m,2 H),1.73-1.67(m,2H),1.61-1.44(m,18H),1.30-1.26(m,60H),0.88-0.86(m,15H). LC-MS(ESI):(M+H)978.4.

[0266] Example 16: Synthesis of aminolipid compound 133 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(CDCl3)δ 4.87-4.83(m,2H),3.64(p,J=7.0 Hz,1H),3.43(q,J=6.2 Hz,2H),3.37(t,J=6.7 Hz,2H),3.12-3.09(m,2H),2.42-2.36(m,2H),2.30-2.20(m,12H),1.94-1.87(m,2 H),1.72-1.66(m,2H),1.62-1.42(m,18H),1.29-1.24(m,62H),0.87-0.85(m,15H). LC-MS(ESI):(M+H)992.4.

[0267] Example 17: Synthesis of aminolipid compound 036 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.95-4.85(m,2H),3.73-3.63(m,1H),3.48(t,J=6.2 Hz,2H),3.41(t,J=6.7 Hz,2H),3.17-3.11(m,2H),2.45(t,J=7.3 Hz,1H),2.41(t,J=7.3 Hz,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). LC-MS(ESI):(M+H)893.9.

[0268] Example 18: Synthesis of aminolipid compound 037 [ka] Following a general synthesis process, aminolipid 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%. 1H NMR(600 MHz,CDCl3)δ 4.08(m,4H),3.73-3.64(m,1H),3.48(t,J=6.1 Hz,2H),3.42(t,J=6.7 Hz,2H),3.15(m,2H),2.45(t,J=7.3 Hz,1H),2.42(t,J=7.3 Hz,1H),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). LC-MS(ESI):(M+H)950.0.

[0269] Example 19: Synthesis of aminolipid compound 038 [ka] Following a general process, aminolipid 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 pale yellow oil with a purity of 95.32% and a yield of 66.2%. 1 H NMR(600 MHz,CDCl3)δ 3.90-3.85(t,J=6.5 Hz,4H),3.70-3.62(m,1H),3.47-3.44(m,2H),3.42-3.40(t,J=6.7 Hz,2H),3.13-3.10(m,2H),2.46-2.42(t,J=7.3 Hz,1H),2.42-2.39(t,J=7.3 Hz,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). LC-MS(ESI):(M+H)950.6.

[0270] Example 20: Synthesis of aminolipid compound 039 [ka] Following a general process, aminolipid compound 039 (0.92 g) was prepared from compound 1522F-7 (2.2 g, 2.59 mmol) and 0.59 g of 130-G as a pale yellow oil with a purity of 95.23% and a yield of 35.3%. 1 H NMR(600 MHz,CDCl3)δ 4.91-4.87(m,2H),3.74-3.71(m,1H),3.45-3.42(m,2H),3.41-3.39(t,J=6.7 Hz,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,2 H),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). LC-MS(ESI):(M+H)1006.4.

[0271] Example 21: Synthesis of aminolipid compound 040 [ka] Following a general process, aminolipid 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%. 1H NMR(600 MHz,CDCl3)δ 4.91-4.87(m,2H),3.78-3.76(m,1H),3.48-3.46(m,2H),3.42-3.40(t,J=6.7 Hz,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,2 H),1.66-1.60(m,4H),1.60-1.56(m,2H),1.53-1.52(m,6H),1.49-1.41(m,4H),1.35-1.21(m,56H),1.06-1.03(t,J=7.1 Hz,6H),0.91-0.88(m,15H). LC-MS(ESI):(M+H)978.3.

[0272] Example 22: Synthesis of aminolipid compound 041 [ka] Following a general process, aminolipid 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 pale yellow oil with a purity of 96.26% and a yield of 44.9%. 1 H NMR(600 MHz,CDCl3)δ 4.93-4.83(m,2H),3.78-3.58(m,1H),3.47(t,J=5.9 Hz,2H),3.41(t,J=6.7 Hz,2H),3.14(m,2H),2.49-2.37(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). LCMS(ESI):(M+H)893.9.

[0273] Example 23: Synthesis of aminolipid compound 042 [ka] Following a general process, aminolipid compound 042 (0.93 g) was prepared from compound 1522F-10 (1.91 g, 2.25 mmol) and 0.47 g of 130-G as a pale yellow oil with a purity of 95.58% and a yield of 41.1%. 1 1H NMR (600 MHz, CDCl3)δ 4.95-4.82(m,2H),3.73-3.62(m,1H),3.50-3.44(m,2H),3.44-3.39(m,2H), 3.16-3.13(m,2H),2.44-2.41(m,4H),2.32-2.29(m,4H),2.29(s,4H),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). LCMS(ESI):(M+H)1006.3.

[0274] Example 24: Synthesis of aminolipid compound 044 [ka] Following a general process, aminolipid compound 044 (0.32 g) was prepared from compound 1522F-11 (1.35 g, 1.49 mmol) and 0.34 g of 130-G as a pale yellow oil with a purity of 95.13% and a yield of 20.2%. 1H NMR(600 MHz,CDCl3)δ 4.95-4.81(m,2H),3.70-3.66(m,1H),3.49-3.46(m,2H),3.43-3.40(t,J=6.7 Hz,2H),3.23-3.07(m,2H),2.46-2.44(t,J=7.3 Hz,1H),2.43-2.40(t,J=7.3 Hz,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). LC-MS(ESI):(M+H)1062.5.

[0275] Example 25: Synthesis of aminolipid compound 045 [ka] Following a general process, aminolipid 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 pale yellow oil with a purity of 97.38% and a yield of 47.2%. 1 H NMR(600 MHz,CDCl3)δ 4.97-4.84(m,2H),3.70-3.66(m,1H),3.49-3.47(t,J=6.1 Hz,2H),3.43-3.40(t,J=6.7 Hz,2H),3.16-3.13(m,2H),2.46-2.44(t,J=7.2 Hz,1H),2.43-2.40(t,J=7.3 Hz,1H),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). LC-MS(ESI):(M+H)893.9.

[0276] Example 26: Synthesis of aminolipid compound 046 [ka] Following a general process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.00(d,J=1.8 Hz,2H),3.99(d,J=1.8 Hz,2H),3.72-3.63(m,1H),3.49-3.46(m,2H),3.43-3.41(t,J=6.7 Hz,2H),3.15(m,2H),2.47-2.44(t,J=7.3 Hz,1H),2.43-2.40(t,J=7.3 Hz,1H),2.34-2.29(m,6H),2.25(s,6H),1.91-1.88(m,4H),1.69-1.61(m,6 H),1.61-1.56(m,2H),1.46(m,4H),1.38-1.16(m,56H),0.96-0.84(m,15H). LC-MS(ESI):(M+H)Found 922.2.

[0277] Example 27: Synthesis of aminolipid compound 047 [ka] Following a general process, aminolipid 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%. 1H NMR(600 MHz,CDCl3)δ 4.94-4.85(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=6.7 Hz,2H),3.16-3.13(m,2H),2.46-2.44(t,J=7.3 Hz,1H),2.43-2.40(t,J=7.3 Hz,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). LC-MS(ESI):(M+H)893.9.

[0278] Example 28: Synthesis of aminolipid compound 048 [ka] Following a general process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.07(t,J=6.8 Hz,4H),3.71-3.62(m,1H),3.48-3.46(t,J=6.2 Hz,2H),3.42-3.40(t,J=6.7 Hz,2H),3.16-3.12(m,2H),2.46-2.43(t,J=7.2 Hz,1H),2.42-2.40(t,J=7.3 Hz,1H),2.33-2.28(m,6H),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). LC-MS(ESI):(M+H)893.9.

[0279] Example 29: Synthesis of aminolipid compound 049 [ka] Following a general process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.09(m,4H),3.73-3.64(m,1H),3.48(t,J=6.1 Hz,2H),3.42(t,J=6.7 Hz,2H),3.15(m,2H),2.46(t,J=7.3 Hz,1H),2.42(t,J=7.3 Hz,1H),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). LC-MS(ESI):(M+H)922.0.

[0280] Example 30: Synthesis of aminolipid compound 122 [ka] Following a general synthesis process, aminolipid compound 122 (0.51 g) was prepared from compound 1522F-1 (1 g, 1.26 mmol) and 0.33 g of 122-G as a pale yellow oil with a purity of 92.94% and a yield of 43%. 1 H NMR in CDCl3δ 4.86-4.85(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). LC-MS(ESI):(M+H)950.5.

[0281] Example 31: Synthesis of aminolipid compound 123 [ka] Following a general synthesis process, aminolipid compound 123 (0.88 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.46 g of 123-G as a pale yellow oil with a purity of 90.77% and a yield of 55%. 1 H NMR in CDCl3δ 4.87-4.85(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). LC-MS(ESI):(M+H)964.5.

[0282] Example 32: Synthesis of aminolipid compound 126 [ka] Following a general synthesis process, aminolipid compound 126 (0.83 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.43 g of 126-G as a pale yellow oil with a purity of 93.73% and a yield of 53%. 1H NMR in CDCl3δ 4.89-4.84(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). LC-MS(ESI):(M+H)950.3.

[0283] Example 33: Synthesis of aminolipid compound 127 [ka] Following a general synthesis process, aminolipid compound 127 (0.75 g) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.46 g of 127-G as a pale yellow oil with a purity of 95.58% and a yield of 48%. 1 H NMR in CDCl3δ 4.88-4.84(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). LC-MS(ESI):(M+H)964.4.

[0284] Example 34: Synthesis of aminolipid compound 128 [ka] Following a general synthesis process, aminolipid compound 128 (0.76 mg) was prepared from compound 1522F-1 (1.3 g, 1.64 mmol) and 0.49 g of 128-G as a pale yellow oily substance with a purity of 94.70% and a yield of 47%. 1 H NMR in CDCl3δ 4.88-4.84(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). LC-MS(ESI):(M+H)978.4.

[0285] Example 35: Synthesis of aminolipid compound 145 [ka] Following a general synthesis process, aminolipid compound 145 (0.24 g) was prepared from compound 1522F-2 (1 g, 1.28 mmol) and 0.28 g of 120-G as a pale yellow oil with a purity of 90.77% and a yield of 20%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.82(m,2H),4.12(d,J=34.5 Hz,2H),3.59-3.44(m,3H),3.32-3.20(m,2H),2.43(dd,J=36.8,28.9 Hz,2H),2.32(s,4H),2.26(td,J=7.6,3.5 Hz,6H),1.60(dd,J=11.6,6.9 Hz,6H),1.54-1.40(m,12H),1.36-1.12(m,56H),0.88(dt,J=17.2,7.0 Hz,15H). LC-MS(ESI):(M+H)908.3

[0286] Example 36: Synthesis of aminolipid compound 146 [ka] Following a general synthesis process, aminolipid compound 146 (0.55 g) was prepared from compound 1522F-2 (1 g, 1.28 mmol) and 0.31 g of 121-G as a pale yellow oil with a purity of 97.28% and a yield of 47%. 1 H NMR(600 MHz,CDCl3)δ 4.86(p,J=6.5 Hz,2H),4.12(d,J=34.8 Hz,2H),3.59-3.45(m,3H),3.25(dd,J=14.6,5.8 Hz,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). LC-MS(ESI):(M+H)922.3.

[0287] Example 37: Synthesis of aminolipid compound 147 [ka] Following a general synthesis process, aminolipid compound 147 (0.92 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g of 122-G as a pale yellow oil with a purity of 92.84% and a yield of 59%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.81(m,2H),4.12(d,J=34.8 Hz,2H),3.59-3.42(m,3H),3.24(dd,J=8.7,6.6 Hz,2H),2.51-2.38(m,2H),2.33-2.22(m,10H),1.64-1.54(m,6H),1.55-1.40(m,12H),1.26(dd,J=8.2,4.6 Hz,60H),0.87(t,J=7.0 Hz,15H). LC-MS(ESI):(M+H)936.3.

[0288] Example 38: Synthesis of aminolipid compound 148 [ka] Following a general synthesis process, aminolipid compound 148 (0.85 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.47 g of 123-G as a pale yellow oil with a purity of 96.59% and a yield of 54%. 1H NMR(600 MHz,CDCl3)δ 4.86(p,J=6.6 Hz,2H),4.12(d,J=34.9 Hz,2H),3.59-3.43(m,3H),3.24(dd,J=8.8,6.6 Hz,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=15.6,9.8 Hz,62H),0.87(t,J=7.0 Hz,15H). LC-MS(ESI):(M+H)950.3.

[0289] Example 39: Synthesis of aminolipid compound 150 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.82(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=40.8,21.6,14.0,6.9 Hz,18H),1.34-1.13(m,56H),0.87(dt,J=14.0,7.0 Hz,15H). LC-MS(ESI):(M+H)922.3.

[0290] Example 40: Synthesis of aminolipid compound 151 [ka] Following a general synthesis process, aminolipid compound 151 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g of 126-G as a pale yellow oil with a purity of 95.27% and a yield of 64%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.82(m,2H),3.77-3.70(m,2H),3.66-3.57(m,1H),3.46-3.38(m,2H),3.25(dd,J=10.9,4.8 Hz,2H),2.64-2.56(m,2H),2.44(dd,J=16.0,8.7 Hz,2H),2.34-2.22(m,10H),1.65-1.37(m,18H),1.27(dd,J=12.0,6.0 Hz,58H),0.91-0.84(m,15H). LC-MS(ESI):(M+H)936.3.

[0291] Example 41: Synthesis of aminolipid compound 152 [ka] Following a general synthesis process, aminolipid compound 152 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.47 g of 127-G as a pale yellow oil with a purity of 95.65% and a yield of 63%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.82(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=15.6,7.5 Hz,2H),2.31-2.22(m,10H),1.66-1.36(m,18H),1.35-1.11(m,60H),0.87(t,J=7.0 Hz,15H). LC-MS(ESI):(M+H)950.4.

[0292] Example 42: Synthesis of aminolipid compound 153 [ka] Following a general synthesis process, aminolipid compound 153 (1.07 g) was prepared from compound 1522F-2 (1.5 g, 1.92 mmol) and 0.58 g of 128-G as a pale yellow oil with a purity of 93.89% and a yield of 58%. 1 H NMR(600 MHz,CDCl3)δ 4.90-4.82(m,2H),3.77-3.70(m,2H),3.65-3.57(m,1H),3.47-3.38(m,2H),3.25(dd,J=9.5,5.8 Hz,2H),2.59(dd,J=9.2,5.5 Hz,2H),2.51-2.40(m,2H),2.31(s,4H),2.26(dt,J=11.7,3.6 Hz,6H),1.64-1.36(m,18H),1.26(d,J=12.4 Hz,62H),0.87(t,J=6.8 Hz,15H). LC-MS(ESI):(M+H)964.4.

[0293] Example 43: Synthesis of aminolipid compound 155 [ka] Following a general synthesis process, aminolipid compound 155 (1.0 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.44 g of 130-G as a pale yellow oil with a purity of 93.39% and a yield of 64%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.82(m,2H),3.64(p,J=6.6 Hz,1H),3.43(q,J=6.0 Hz,2H),3.38(t,J=6.7 Hz,2H),3.28-3.18(m,2H),2.52-2.34(m,4H),2.34-2.21(m,10H),1.96-1.84(m,2H),1.52(ddt,J=21.3,13.8,6.6 Hz,18H),1.35-1.07(m,56H),0.87(dt,J=7.2,6.2 Hz,15H). LC-MS(ESI):(M+H)936.4.

[0294] Example 44: Synthesis of aminolipid compound 156 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.86(p,J=5.7 Hz,2H),3.64(p,J=6.8 Hz,1H),3.43(q,J=6.1 Hz,2H),3.38(t,J=6.7 Hz,2H),3.22(dd,J=15.6,11.5 Hz,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). LC-MS(ESI):(M+H)950.3.

[0295] Example 45: Synthesis of aminolipid compound 157 [ka] Following a general synthesis process, aminolipid 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%. 1H NMR(600 MHz,CDCl3)δ 4.89-4.81(m,2H),3.64(p,J=6.6 Hz,1H),3.43(qd,J=6.0,1.1 Hz,2H),3.38(t,J=6.7 Hz,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). LC-MS(ESI):(M+H)964.4.

[0296] Example 46: Synthesis of aminolipid compound 158 [ka] Following a general synthesis process, aminolipid compound 158 (0.96 g) was prepared from compound 1522F-2 (1.3 g, 1.67 mmol) and 0.54 g of 133-G as a pale yellow oil with a purity of 97.59% and a yield of 59%. 1 H NMR(600 MHz,CDCl3)δ 4.85(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). LC-MS(ESI):(M+H)978.4.

[0297] Example 47: Synthesis of aminolipid compound 167 [ka] Following a general synthesis process, aminolipid compound 167 (0.45 g) was prepared from compound 1522F-3 (1 g, 1.24 mmol) and 0.27 g of 120-G as a pale yellow oil with a purity of 92.68% and a yield of 39%. 1 H NMR(600 MHz,CDCl3)δ 4.92-4.79(m,2H),4.13(s,0.6H),4.10(s,1.4H),3.61-3.42(m,3H),3.13-3.04(t,J=6.0 Hz,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). LC-MS(ESI):(M+H)936.2.

[0298] Example 48: Synthesis of aminolipid compound 168 [ka] Following a general synthesis process, aminolipid compound 168 (0.65 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.39 g of 121-G as a pale yellow oil with a purity of 91.4% and a yield of 42%. 1 H NMR(600 MHz,CDCl3)δ 4.97-4.79(m,2H),4.16(s,0.6H),4.14(s,1.4H),3.62-3.44(m,3H),3.11(t,J=6.0 Hz,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). LC-MS(ESI):(M+H)950.1.

[0299] Example 49: Synthesis of aminolipid compound 169 [ka] Following a general synthesis process, aminolipid compound 169 (0.58 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 122-G as a pale yellow oil with a purity of 91.97% and a yield of 37%. 1 H NMR(600 MHz,CDCl3)δ 4.93-4.81(m,2H),4.16(s,0.6H),4.13(s,1.4H),3.63-3.48(m,3H),3.16-3.07(t,J=6.0 Hz,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=6.8 Hz,15H). LC-MS(ESI):(M+H)964.1.

[0300] Example 50: Synthesis of aminolipid compound 170 [ka] Following a general synthesis process, aminolipid compound 170 (0.41 g) was prepared from compound 1522F-3 (1 g, 1.24 mmol) and 0.35 g of 123-G as a pale yellow oil with a purity of 92.56% and a yield of 34%. 1 H NMR(600 MHz,CDCl3)δ 4.94-4.84(m,2H),4.16(s,0.6H),4.13(s,1.4H),3.62-3.48(m,3H),3.15-3.08(t,J=6.0 Hz,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=7.0 Hz,15H). LC-MS(ESI):(M+H)978.1.

[0301] Example 51: Synthesis of aminolipid compound 171 [ka] Following a general synthesis process, aminolipid compound 171 (0.52 g) was prepared from compound 1522F-3 (1 g, 1.24 mmol) and 0.27 g of 149-G as a pale yellow oil with a purity of 91.21% and a yield of 46%. 1 H NMR(600 MHz,CDCl3)δ 4.92-4.79(m,2H),3.74(q,J=7.1 Hz,2H),3.66-3.57(m,1H),3.44(q,J=6.6 Hz,2H),3.10-3.06(m,2H),2.60(t,J=7.0 Hz,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). LC-MS(ESI):(M+H)936.1.

[0302] Example 52: Synthesis of aminolipid compound 173 [ka] Following a general synthesis process, aminolipid compound 173 (0.42 g) was prepared from compound 1522F-3 (1 g, 1.24 mmol) and 0.32 g of 126-G as a pale yellow oil with a purity of 92.16% and a yield of 36%. 1 H NMR(600 MHz,CDCl3)δ 4.91-4.81(m,2H),3.74(q,J=7.1 Hz,2H),3.66-3.57(m,1H),3.43(q,J=6.6 Hz,2H),3.13-3.01(m,2H),2.60(t,J=7.0 Hz,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). LC-MS(ESI):(M+H)964.1.

[0303] Example 53: Synthesis of aminolipid compound 174 [ka] Following a general synthesis process, aminolipid compound 174 (0.53 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.45 g of 127-G as a pale yellow oil with a purity of 97.18% and a yield of 34%. 1 H NMR(600 MHz,CDCl3)δ 4.92-4.81(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=7.0 Hz,2H),2.58(t,J=7.1 Hz,0.5H),2.47-2.41(t,J=6.9 Hz,1.5H),2.38-2.28(m,10H),1.71-1.40(m,22H),1.38-1.17(m,60H),0.91(t,J=7.0 Hz,15H). LC-MS(ESI):(M+H)978.2.

[0304] Example 54: Synthesis of aminolipid compound 175 [ka] Following a general synthesis process, aminolipid 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%. 1 H NMR(600 MHz,CDCl3)δ 4.89-4.81(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=7.0 Hz,2H),2.54(t,J=7.1 Hz,0.5H),2.43(t,J=6.9 Hz,1.5H),2.36-2.24(m,10H),1.69-1.36(m,22H),1.35-1.10(m,62H),0.88(t,J=7.0 Hz,15H). LC-MS(ESI):(M+H)992.2.

[0305] Example 55: Synthesis of aminolipid compound 177 [ka] Following a general synthesis process, aminolipid compound 177 (0.6 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 130-G as a pale yellow oil with a purity of 92.65% and a yield of 39%. 1 H NMR(600 MHz,CDCl3)δ 4.94-4.80(m,2H),3.69-3.56(m,1H),3.48-3.41(m,2H),3.41-3.35(m,2H),3.07(t,J=7.0 Hz,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). LC-MS(ESI):(M+H)964.2.

[0306] Example 56: Synthesis of aminolipid compound 178 [ka] Following a general synthesis process, aminolipid compound 178 (0.43 g) was prepared from compound 1522F-3 (1.3 g, 1.61 mmol) and 0.42 g of 131-G as a pale yellow oil with a purity of 94.43% and a yield of 27%. 1 H NMR(600 MHz,CDCl3)δ 4.90-4.84(m,2H),3.70-3.58(m,1H),3.48-3.43(m,2H),3.43-3.38(m,2H),3.08(t,J=7.0 Hz,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). LC-MS(ESI):(M+H)978.2.

[0307] Comparative Example 1: Synthesis of Compound 118 [ka] Following a general synthesis process, compound 118 (0.85 g) was prepared from 1.3 g of compound 1522F-1 (1.3 g, 1.64 mmol) and 0.31 g of 118-G as a colorless oil with a purity of 90.77% and a yield of 57%. 1 H NMR in(CDCl3)δ 4.86-4.85(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). LC-MS(ESI):(M+H)920.5.

[0308] Biological tests Compounds 118 and 1004 used in the biological studies of this disclosure have the following structures: [ka]

[0309] Compound 118 can be prepared according to the synthesis method of Compound 10 (Example 9) described in Comparative Example 1 or in Chinese Patent No. 107922364A, and Compound 1004 can be prepared according to the synthesis method of Compound 7 (Example 7) in Chinese Patent No. 114127044A.

[0310] Experimental Example 1: Preparation of lipid nanoparticles containing luciferase mRNA (Fluc mRNA)

[0311] (1) Formulation: The specified amount of Fluc mRNA stock solution, 0.2 M sodium acetate buffer, and DEPC water were added to a container and mixed thoroughly to obtain the aqueous phase. The aminolipid compounds, helper lipids, structural lipids, and PEG-lipids of this disclosure were separately dissolved in anhydrous ethanol at concentrations of 20 mg / mL, 10 mg / mL, 20 mg / mL, and 25 mg / mL, respectively, to prepare the respective solutions. The above four solutions were pipettered in a molar ratio of aminolipid compound:DSPC:CHO-HP:PEG2000-DMG of 48:10:40.5:1.5 and thoroughly mixed to prepare the alcohol phase.

[0312] (2) Encapsulation: Using a microfluidic preparation device (MPE-L2), the aqueous phase:alcohol phase was injected into a microfluidic chip at a flow rate of 12 mL / min:4 mL / min, and encapsulation was performed at a flow rate of 9 mL / min:3 mL / min to obtain mRNA-encapsulated lipid nanoparticles (mRNA-LNP).

[0313] (3) Dialysis: The product from step (2) was placed in a dialysis bag and then placed in a replacement Tris buffer-8% sucrose solution to remove components such as residual ethanol and unorganized lipids. Dialysis was performed at room temperature for 2 hours with magnetic agitation while protecting from light (the dialysate was changed every hour).

[0314] (4) The product from step (3) was sterilized by passing it through a 0.22 μm microporous membrane and then packaged. Lipid nanoparticle formulations containing fluc mRNA were prepared with a fluc mRNA concentration of 0.2 μg / μL, a mass ratio of fluc mRNA to lipid of 1:10, a particle size of 80–130 nm, and an encapsulation efficiency of 80% or more.

[0315] Experimental Example 2: Performance Evaluation of In Vivo Delivery of Lipid Nanoparticles Animal preparation: Female BALB / c mice aged 6-8 weeks were selected and housed in SPF-grade housing facilities. Animal studies were conducted strictly in accordance with national healthcare guidelines and animal ethics requirements.

[0316] In vivo delivery: Before injecting the test LNP formulation, the formulation sample was thoroughly mixed by gently and repeatedly inverting the LNP formulation. The corresponding amount of formulation sample was aspirated into a 1 ml insulin syringe, and the LNP formulation was injected via tail vein injection (IV), with two formulations injected into three mice. 75 μL of the luciferase mRNA (Fluc mRNA)-encapsulated lipid nanoparticle formulation prepared in Experimental Example 1 was injected into each mouse.

[0317] Six hours after injection of the LNP preparation, mice were injected with 200 μL of D-luciferin luciferase expression substrate (catalog number 122799, manufacturer: Perkin Elmer). After substrate injection, the mice were anesthetized by injecting 200 μL of 2.5% aveltin, and the injection time of the luciferase expression 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 body and organs were observed using an in vivo imaging system (IVIS).

[0318] Table 4 shows the fluorescence expression intensities induced by lipid nanoparticles containing luciferase mRNA (Fluc mRNA) with representative aminolipid compounds, using aminolipid compound 118 as a control. [Table 4] JPEG0007900866000098.jpg74149 Table 5 shows the spleen delivery / total delivery ratio (spleen / total flux) for fluorescence expression intensity induced by lipid nanoparticles containing luciferase mRNA (Fluc mRNA) of representative aminolipid compounds for compound 118. [Table 5] JPEG0007900866000100.jpg68149 As can be seen from Table 5, aminolipid compounds other than aminolipid compounds 145, 146, 150, 151, 156, and 157 showed a stronger delivery preference to the spleen compared to aminolipid compound 118.

[0319] Experimental Example 3: Evaluation of the delivery efficiency of lipid nanoparticles to immune cell populations in different tissues and organs. C57BL / 6 mice (3 mice per group) were intravenously injected with eGFP mRNA encapsulating LNPs. Six hours later, the mice were dissected, and the spleen, axillary lymph nodes, peripheral blood, bone marrow, lungs, and liver were collected to prepare single-cell suspensions from various tissues and organs. After blocking with an Fc blocking antibody and surface staining with a fluorescently labeled antibody, samples were analyzed by flow cytometry to analyze the expression levels of eGFP in various immune cell populations, confirming the delivery efficiency of lipid nanoparticles containing different aminolipid compounds to immune cell populations in different tissues and organs.

[0320] The test results are shown in Figures 1A to 1F. As can be seen from the figures, lipid nanoparticles containing aminolipid compounds 130, 131, and 133 exhibit clear delivery preference to immune cells in various tissues and organs.

[0321] In addition to what is described herein, various modifications to this disclosure will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to be included within the scope of the attached claims.

Claims

1. An aminolipid compound having the structure of formula (II), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. 【Chemistry 1】 (In the formula, Z ７ and Z ８ Each of them is independently -(C=O)O- or -O(C=O)-, A3 is a linear C1-C4 alkylene, A4, A6, and A7 are each independently C1-C12 alkylenes. R １ and R ２ Each of them is independently a C1-C18 alkyl, R ３ It is a C1-C18 alkyl group, R ４ and R ５ Each of these is independently a C1-C20 alkyl group.

2. A ４ is a linear C １ -C ６ alkylene, the amino lipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

3. The aminolipid compound according to claim 1 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein A4 is a linear C1-C3 alkylene.

4. A ３ is a straight chain C ２ -C ４ An aminolipid compound according to claim 1, which is alkylene, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

5. The aminolipid compound according to claim 1 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein A3 is a linear C3 or C4 alkylene.

6. Z ７ and Z ８ The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein each of the isopropyl alcohols is -(C=O)O-.

7. A ６ and A ７ Each of them operates independently, linear C ３ , C ４ , C ５ , C ６ , C ７ , C ８ , C ９ , C １０ or C １１ An aminolipid compound according to claim 1, which is alkylene, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

8. The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein A6 and A7 are each independently linear C5-C11 alkylenes.

9. R １ and R ２ Each of them operates independently, C １ -C ４ An aminolipid compound according to claim 1, which is alkyl, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

10. The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R1 and R2 are each independently C1 or C2 alkyl.

11. R ３ C １ , C ２ , C ３ , C ４ , C ５ , C ６ , C ７ , C ８ , C ９ , C １０ , C １１ or C １２ An aminolipid compound according to claim 1, which is alkyl, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

12. The aminolipid compound according to claim 1, wherein R3 is a linear C4-C8 alkyl group, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

13. R ４ and R ５ Each of them operates independently, branch C ３ -C ２０ An aminolipid compound according to claim 1, which is alkyl, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

14. The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R4 and R5 are each independently a branched C5-C20 alkyl group.

15. Z ７ and Z ８ Each of them independently is -(C=O)O- or -O(C=O)-, A ３ However, linear C ２ -C ４ It is alkylene, A ４ However, linear C １ -C ３ It is alkylene, A ６ and A ７ Each of them operates independently, linear C ３ , C ４ , C ５ , C ６ , C ７ , C ８ , C ９ , C １０ or C １１ It is alkylene, R １ and R ２ Each of them operates independently, C １ -C ４ It is alkyl, R ３ However, C １ , C ２ , C ３ , C ４ , C ５ , C ６ , C ７ , C ８ , C ９ , C １０ , C １１ or C １２ It is alkyl, R ４ and R ５ Each of them operates independently, branch C ３ -C ２０ It is alkyl. The aminolipid compound described in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

16. Z ７ and Z ８ Each of them is independent of -(C=O)O-, A ３ is a linear C ３ -C ４ alkylene, A ４ However, linear C １ -C ３ It is alkylene, A ６ and A ７ are each independently a linear C ５ -C １１ alkylene, R １ and R ２ Each of them operates independently, C １ or C ２ It is alkyl, R ３ However, linear C ４ -C ８ It is alkyl, R ４ and R ５ Each of them operates independently, branch C ５ -C ２０ It is alkyl. The aminolipid compound described in claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

17. The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein the aminolipid compound has one of the structures shown below. Table 1

18. Lipid nanoparticles comprising the aminolipid compound described in any one of claims 1 to 17.

19. The lipid nanoparticles according to claim 18, further comprising a biologically active ingredient.

20. The lipid nanoparticle according to claim 19, wherein the biologically active component is nucleic acid.

21. A pharmaceutical composition comprising an aminolipid compound according to any one of claims 1 to 17 or lipid nanoparticles containing the aminolipid compound, and a pharmaceutically acceptable carrier, diluent, or excipient.

22. Lipid nanoparticles according to claim 18 for use as a vehicle for an active ingredient.

23. The lipid nanoparticles according to claim 22, wherein the active ingredient is a biologically active ingredient.

24. The lipid nanoparticle according to claim 22, wherein the active ingredient is nucleic acid.

25. A pharmaceutical product comprising the lipid nanoparticles described in claim 18, or a pharmaceutical composition comprising the lipid nanoparticles and a pharmaceutically acceptable carrier, diluent, or excipient.

26. The pharmaceutical product according to claim 25, wherein the pharmaceutical product is for use in gene therapy, gene vaccination or protein replacement therapy, antisense therapy or interfering RNA therapy.

27. A pharmaceutical product comprising, for nucleic acid transfer, the lipid nanoparticles described in claim 18, or a pharmaceutical composition comprising the lipid nanoparticles and a pharmaceutically acceptable carrier, diluent, or excipient.

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