Amino lipid compound and use thereof

Abstract

The present disclosure relates to an amino lipid compound and use thereof. The present disclosure also relates to an empty lipid nanoparticle, a loaded lipid nanoparticle, and a pharmaceutical composition that comprise the amino lipid compound, and use thereof.

Description

Amino lipid compounds and their applications Technical Field

[0001] This disclosure relates to an aminolipid compound and its applications. This disclosure also relates to lipid nanoparticles and pharmaceutical compositions containing said aminolipid compound, and their uses. Background Technology

[0002] Gene therapy refers to the introduction of exogenous normal genes into target cells to achieve therapeutic effects or induce an immune response. However, gene therapy drugs face several challenges, especially nucleic acid drugs. Naked nucleic acids are difficult to directly introduce into cells and are easily degraded by nucleases in the cytoplasm, making vector delivery technology particularly crucial. The use of lipid nanoparticles to deliver nucleic acids has been widely adopted; however, although a large number of lipid nanoparticles have been developed, their safety, efficacy, and specificity remain insufficient.

[0003] Therefore, in order to improve the efficacy of nucleic acid drugs, it is of great research significance and practical need to develop more different lipid nanoparticles, especially the development of amino lipid compounds for formulating lipid nanoparticles, as well as related preparation methods and applications. Summary of the Invention

[0004] On the one hand, this disclosure provides aminolipid compounds as shown in formula (I), formula (II) or formula (III):

[0005] Or its pharmaceutically acceptable salt, or its stereoisomer,

[0006] in,

[0007] X1 and X2 are independently selected from O or S;

[0008] R1 is Where R ’ H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C2-C6 heteroalkenyl; R ” H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, or

[0009] X3 and X4 are independently selected from O or S;

[0010] R M H, halogen, -OH, -R * -N(R) * )2, -CN, -N3, -C(=O)OH, -C(=O)OR *-OC(=O)R * -OR * -SR * -S(=O)R * -S(=O)OR * -S(=O)2OR * -NO2, -S(=O)2N(R) * )2、-N(R * )S(=O)2R * -NH(CH2) t1 N(R * 2、-NH(CH2) p1 O(CH2) q1 N(R * 2、-NH(CH2) s1 OR * -N((CH2) s1 OR * )2、-N(R * -Carbon ring, -N(R) * - Heterocyclic rings, -N(R) * -Aryl, -N(R) * )-Heteroaryl, -N(R * (CH2) t1 -Carbon ring, -N(R) * (CH2) t1 -heterocyclic, -N(R) * (CH2) t1 -Aryl, -N(R) * (CH2) t1 - Heteroaryl, carbocyclic, heterocyclic, aryl or heteroaryl;

[0011] Each R * It is independently H, C1-C3 alkyl, or C2-C3 alkenyl;

[0012] Each t1 is independently 1, 2, 3, 4 or 5;

[0013] p1 is 1, 2, 3, 4 or 5;

[0014] q1 is 1, 2, 3, 4, or 5;

[0015] Each s1 is independently 1, 2, 3, 4 or 5;

[0016] R N It is H or C1-C3 alkyl;

[0017] r is 0 or 1;

[0018] n is any integer from 0 to 10;

[0019] L3 is a bond, an optional substituted C1-C6 alkylene group, or an optional substituted C1-C6 heteroalkylene group;

[0020] R2 is H, a C1-C6 hydrocarbon group, or a C1-C6 heterohydrocarbon group;

[0021] L1 and L2 are independently selected from bonded, optionally substituted C1-C3 alkylene or optionally substituted C2-C3 alkenyl groups;

[0022] A1 and A2 are independently selected from C1 to C2. 10 Alkylene or C1-C 10 Heteroalkyl;

[0023] A3 and A4 are independently selected from C1-C6 alkylene, C1-C6 heteroalkylene, or bonded;

[0024] R3 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5);

[0025] R4 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6);

[0026] Each M1 is independently C1~C 24 Hydroxyl group;

[0027] Each M2 is independently C1 to C 12 Hydroxyl group;

[0028] Each M a Independently C1~C 24 hydrocarbon group;

[0029] Each M b Independently C1~C 15 hydrocarbon group;

[0030] Z1 and Z2 are independently selected from -C(=O)O- or -OC(=O)-;

[0031] Each R a Independently, it is an H or C1-C8 hydrocarbon group;

[0032] Each R5 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ;

[0033] Each R6 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ;

[0034] Each M3 is independently C3~C 12 Hydroxyl group;

[0035] Each M4 is independently a C1–C8 hydrocarbon group;

[0036] Each M c Independently C3~C 12 hydrocarbon group;

[0037] Each M d Independently composed of C1–C8 hydrocarbon groups;

[0038] Each Y is an independent 3-7 member carbon ring.

[0039] In some implementations, X1 and X2 are 0.

[0040] In some implementations, Z1 and Z2 are -C(=O)O-.

[0041] In some embodiments, the aminolipid compound is as shown in formula (I), wherein R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is -C(R a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6). In some implementations, R3 is C1 to C 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0042] In some embodiments, the amino lipid compound is as shown in formula (I), wherein R3 is -CH(OR5)2 and R4 is -CH(OR6)2.

[0043] In some embodiments, the aminolipid compound is as shown in formula (I), wherein X1 and X2 are O, and R3 is C1 to C2. 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0044] In some embodiments, the aminolipid compound is as shown in formula (I), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0045] In some embodiments, the aminolipid compound is as shown in formula (I), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0046] In some embodiments, the aminolipid compound has a structure as shown in formula (IV):

[0047] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in this paper for equation (I).

[0048] In some embodiments, the aminolipid compound is as shown in formula (II), wherein R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is -C(R a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6). In some implementations, R3 is C1 to C 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0049] In some embodiments, the aminolipid compound is as shown in formula (II), wherein R3 is -C(R a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is C1~C 24 hydrocarbon group, -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a(SR6)(OR6). In some implementations, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group or -CH(OR6)2.

[0050] In some embodiments, the aminolipid compound is as shown in formula (II), wherein R3 is -CH(OR5)2 and R4 is -CH(OR6)2.

[0051] In some embodiments, the aminolipid compound is as shown in formula (II), wherein X1 and X2 are O, and R3 is C1 to C2. 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0052] In some embodiments, the aminolipid compound is as shown in formula (II), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0053] In some embodiments, the aminolipid compound is as shown in formula (II), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0054] In some embodiments, the aminolipid compound is as shown in formula (II), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0055] In some embodiments, the amino lipid compound is as shown in formula (II), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group or -CH(OR6)2.

[0056] In some embodiments, the aminolipid compound has a structure as shown in formula (V):

[0057] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in equation (II) in this paper.

[0058] In some embodiments, the aminolipid compound is as shown in formula (III), wherein R3 is C1 to C2. 24 Hydrocarbon group, R4 is C1 to C2. 24 Hydrocarbon group.

[0059] In some embodiments, the amino lipid compound is as shown in formula (III), wherein Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a (OR6)2. In some implementations, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0060] In some embodiments, the amino lipid compound is as shown in formula (III), wherein Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0061] In some embodiments, the aminolipid compound is as shown in formula (III), wherein R3 is C1 to C2. 24 The hydrocarbon group, R4 is -CH(OR6)2, and A1 and A2 are independently selected from C4 to C9 alkylene groups.

[0062] In some embodiments, the amino lipid compound is as shown in formula (III), wherein R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 and A2 are independently selected from C4 to C9 alkylene compounds.

[0063] In some embodiments, the aminolipid compound is as shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a (OR6)2. In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0064] In some embodiments, the aminolipid compound has a structure as shown in formula (VI):

[0065] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in this paper for equation (III).

[0066] In some embodiments, the aminolipid compound is as shown in formula (III), wherein X1 and X2 are O, and R3 is C1 to C2. 24 The hydrocarbon group, R4 is -CH(OR6)2, and A1 and A2 are independently selected from C4 to C9 alkylene groups.

[0067] In some embodiments, the amino lipid compound is as shown in formula (III), wherein X1 and X2 are O, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 and A2 are independently selected from C4 to C9 alkylene compounds.

[0068] In some embodiments, the aminolipid compound is as shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 The hydrocarbon group, R4 is -CH(OR6)2, wherein A1 and A2 are independently selected from C4 to C9 alkylene groups.

[0069] In some embodiments, the amino lipid compound is as shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 and A2 are independently selected from C4 to C9 alkylene groups.

[0070] On the one hand, this disclosure provides empty lipid nanoparticles containing the amino lipid compounds in any of the above embodiments.

[0071] On the one hand, this disclosure provides loaded lipid nanoparticles, which include the above-mentioned empty lipid nanoparticles and active ingredients.

[0072] In another aspect, this disclosure provides a pharmaceutical composition comprising the aforementioned loaded lipid nanoparticles and a pharmaceutically acceptable carrier, diluent, or excipient.

[0073] Furthermore, this disclosure provides the use of the aforementioned empty lipid nanoparticles in the preparation of delivery carriers.

[0074] Furthermore, this disclosure provides a method for delivering an active ingredient to cells, tissues, or organs, including:

[0075] This allows the aforementioned lipid-loaded nanoparticles to come into contact with cells, tissues, or organs.

[0076] Furthermore, this disclosure provides a method for generating polypeptides and / or proteins of interest in mammalian cells, comprising:

[0077] The above-mentioned lipid-loaded nanoparticles containing mRNA are provided, and cells are brought into contact with the lipid-loaded nanoparticles.

[0078] Furthermore, this disclosure provides the use of the aforementioned lipid-loaded nanoparticles or the aforementioned pharmaceutical compositions in the treatment and / or prevention of diseases or conditions.

[0079] Furthermore, this disclosure provides methods for treating and / or preventing diseases or conditions in mammals in need, including:

[0080] Administer to mammals a therapeutic and / or preventative amount of the above-described loaded lipid nanoparticles or the above-described pharmaceutical composition.

[0081] Furthermore, the lipid-loaded nanoparticles or the pharmaceutical compositions described above provided in this disclosure can be used as a drug.

[0082] Furthermore, this disclosure provides a medicine comprising the aforementioned loaded lipid nanoparticles or the aforementioned pharmaceutical composition. Attached Figure Description

[0083] Figure 1 shows the detection results of whole-body fluorescence signals in mice. In Figure 1, "1-01" represents lipid nanoparticles containing aminolipid compounds numbered 1-01 in Table 1, and so on.

[0084] Figures 2A-2F show the detection results of fluorescence signals in various organs of mice.

[0085] Figure 3 shows the results of hEPO concentration detection in rat serum 6 hours after drug administration.

[0086] Figure 4 shows the results of hEPO concentration detection in rat serum 24 hours after drug administration. Detailed Implementation

[0087] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. The term "technique" as used herein refers to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0088] As used herein, the terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations thereof, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0089] As used herein, the term "hydrocarbon group" refers to the group remaining after an aliphatic hydrocarbon loses one hydrogen atom, including straight-chain or branched, saturated or unsaturated hydrocarbon groups. Hydrocarbon groups include, but are not limited to, alkyl, alkenyl, and alkynyl groups. Preferably, the hydrocarbon group has 1 to 24 carbon atoms (C1-C2). 24Hydrocarbon group), for example, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms (C1, C2, C3, ... C2). 21 C 22 C 23 Or C 24 Hydrocarbon groups). Examples of hydrocarbon groups include, but are not limited to, C1-C1 groups. 24 Hydrocarbon group, C1-C 22 Hydrocarbon group, C1-C 20 Hydrocarbon group, C1-C 18 Hydrocarbon group, C1-C 16 Hydrocarbon group, C1-C 12 Hydrocarbon group, C1-C 10 Hydrocarbon groups, C1-C8 hydrocarbon groups, C1-C7 hydrocarbon groups, C1-C6 hydrocarbon groups, C1-C5 hydrocarbon groups, C1-C3 hydrocarbon groups, C1-C2 hydrocarbon groups, C2-C8 hydrocarbon groups, C2-C4 hydrocarbon groups, C4-C8 hydrocarbon groups, C4-C9 hydrocarbon groups, C5-C8 hydrocarbon groups, C1-C4 hydrocarbon groups, C3-C8 hydrocarbon groups, C3 hydrocarbon groups, C4 hydrocarbon groups, C5 hydrocarbon groups, C6 hydrocarbon groups, C7 hydrocarbon groups, and C8 hydrocarbon groups. Unless otherwise expressly stated in this specification, hydrocarbon groups are optionally substituted, and substituents are defined below in relation to the definition of "optionally substituted". In some embodiments, hydrocarbon groups have zero branches (i.e., straight chains), one branch, two branches, or more branches.

[0090] As used herein, the term "hydroalkylene group" refers to the divalent group remaining after a hydrocarbon group as defined above loses one more hydrogen atom. Unless otherwise expressly stated in this specification, the hydroalkylene group may also be substituted.

[0091] As used herein, the term "alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group. Preferably, the alkyl group has 1 to 24 carbon atoms (C1-C2). 24 Alkyl groups, for example, having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms (C1, C2, C3, ... C2). 21 C 22 C 23 Or C 24 Alkyl groups. Examples of alkyl groups include, but are not limited to, C1-C1 alkyl groups. 24 Alkyl, C1-C 22 Alkyl, C1-C 20 Alkyl, C1-C 18 Alkyl, C1-C 16 Alkyl, C1-C 12 Alkyl, C1-C 10Alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 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, C3-C8 alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, and tridecyl-7-yl. Unless otherwise expressly stated in this specification, alkyl groups may be optionally substituted.

[0092] As used herein, the term "alkylene" refers to a divalent group remaining after the alkyl group as defined above loses one more hydrogen atom. Alkylenes may also be optionally substituted, unless otherwise expressly stated in this specification.

[0093] As used herein, the term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbon group containing one or more double bonds (C=C). Preferably, the alkenyl group has 2 to 24 carbon atoms (C2-C4). 24 Alkenyl), for example, having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms (C2, C3, C4, ... C2). 21 C 22 C 23 Or C 24 Alkenyl groups (including but not limited to C2-C4 groups) and having 1, 2, 3, 4 or more double bonds. 24 alkenyl, C2-C 22 alkenyl, C2-C 20 alkenyl, C2-C 18 alkenyl, C2-C 16 alkenyl, C2-C 12 alkenyl, C2-C 10 Alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl, C4-C8 alkenyl, C4-C9 alkenyl, C5-C8 alkenyl, having 1, 2, 3, 4 or more double bonds. Some more specific examples include, but are not limited to, vinyl, propenyl, but-1-enyl, but-2-enyl, pent-1-enyl, pent-2-enyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hep-1-enyl, hep-2-enyl, hep-3-enyl, oct-1-enyl, oct-2-enyl, oct-3-enyl, non-1-enyl, non-2-enyl and non-3-enyl. In some preferred embodiments, the alkenyl group has one double bond. Unless otherwise expressly stated in this specification, the alkenyl group is optionally substituted.

[0094] As used herein, the term "alkenyl" refers to the divalent group remaining after an alkenyl group as defined above loses one more hydrogen atom. Unless otherwise expressly stated in this specification, the alkenyl group may also be optionally substituted.

[0095] As used herein, the term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbon group containing one or more triple bonds (C≡C). Preferably, the alkynyl group has 2 to 24 carbon atoms (C2-C4). 24 Alkyne group, for example, having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms (C2, C3, C4, ... C2). 21 C 22 C 23 Or C 24 The alkynyl group (containing one, two, three, four, or more triple bonds) includes, but is not limited to, C2-C4 groups. 24 Alkyne group, C2-C 22 Alkyne group, C2-C 20 Alkyne group, C2-C 18 Alkyne group, C2-C 16 Alkyne group, C2-C 12 Alkyne group, C2-C 10 The ynyl group, C2-C8 ynyl group, C2-C7 ynyl group, C2-C6 ynyl group, C2-C4 ynyl group, C2-C3 ynyl group, C4-C8 ynyl group, C4-C9 ynyl group, and C5-C8 ynyl group, having 1, 2, 3, 4, or more triple bonds. Some more specific examples include, but are not limited to, ethynyl, propynyl, but-1-ynyl, but-2-ynyl, pent-1-ynyl, pent-2-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hep-1-ynyl, hep-2-ynyl, hep-3-ynyl, oct-1-ynyl, oct-2-ynyl, oct-3-ynyl, non-1-ynyl, non-2-ynyl, and non-3-ynyl. In some preferred embodiments, the ynyl group has one triple bond. Unless otherwise expressly stated in this specification, the ynyl group is optionally substituted.

[0096] As used herein, the term "ynynyl" refers to the divalent group remaining after the ynyl group as defined above loses one more hydrogen atom. Unless otherwise expressly stated in this specification, the ynynyl group may also be optionally substituted.

[0097] As used herein, the term "heteroalkyl group" or its subordinate concepts (such as heteroalkyl, heteroalkenyl, heteroynyl, etc.) refers to a stable straight-chain or branched hydrocarbon group or combination thereof, consisting of a certain number of carbon atoms and at least one heteroatom. A heteroatom refers to an atom other than carbon and hydrogen. In some embodiments, the heteroalkyl group contains one, two, three, or more heteroatoms. In some embodiments, the heteroalkyl group contains one or more (e.g., two or three) identical heteroatoms, or contains multiple (e.g., two or three) different heteroatoms. Preferably, the heteroatom is selected from O, N, or S. Examples of heteroalkyl groups include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-CH2-O-CH2-CH3, -CH2-(CH2)3-O-(CH2)5-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-S-CH3, -CH=CH-O-CH3, -CH2-CH=N-OCH3, –CH=CH-N(CH3)-CH3, and -CH2-NH-OCH3. Unless otherwise expressly stated in this specification, heteroalkyl groups or their subordinate concepts (such as heteroalkyl, heteroalkenyl, heteroynyl, etc.) may be optionally substituted.

[0098] As used herein, the term "heteroalkyl" or its subordinate concepts (such as heteroalkyl, heteroenyl, heteroynyl, etc.) refers to the divalent group remaining after a heteroalkyl group as defined above loses one more hydrogen atom. Unless otherwise expressly stated in this specification, heteroalkyl or its subordinate concepts (such as heteroalkyl, heteroenyl, heteroynyl, etc.) may be optionally substituted.

[0099] As used herein, the term "carbocyclic" or "carbocyclic group" refers to a optionally substituted monocyclic or polycyclic system comprising one or more rings consisting of carbon atoms. The ring can be ternary, quaternary, pentaneous, hexanal, septaneous, octaneous, nonanal, decanal, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, or twentieth-membered rings. For example, "4-7-membered carbocyclic" means a carbocyclic ring comprising a monocyclic ring having 4-7 ring carbon atoms. The carbocyclic ring may include one or more carbon-carbon double or triple bonds and may be a non-aromatic or aromatic ring (e.g., cycloalkyl, cycloalkenyl, or aryl). Examples of carbocyclic rings include cyclopropyl, cyclopentyl, cyclohexyl, phenyl, naphthyl, and 1,2-dihydronaphthyl. The "carbocyclic" or "carbocyclic group" may optionally be substituted with one or more substituents, as defined below for "optionally substituted."

[0100] As used herein, the terms "heterocyclic," "heterocyclic group," or "subheterocyclic group" refer to a cyclic group having a cyclic structure and containing one or more heteroatoms in the cyclic atom. In some embodiments, the cyclic atom contains one or more identical or different heteroatoms. In some embodiments, the one or more heteroatoms in the cyclic atom are selected from N, O, or S. The "heterocyclic," "heterocyclic group," or "subheterocyclic group" disclosed herein are saturated or unsaturated. In some embodiments, the "heterocyclic," "heterocyclic group," or "subheterocyclic group" comprises a monocyclic, bicyclic, or polycyclic ring. In some embodiments, the "heterocyclic," "heterocyclic group," or "subheterocyclic group" is a 4-10 membered heterocycle, such as a 4-7 membered heterocycle or a 5-7 membered heterocycle. Preferably, in some embodiments, the heterocycle is a 4-10 membered heterocycle that may be optionally substituted, wherein the cyclic atom contains 1, 2, 3, 4, 5, or 6 heteroatoms selected from N, O, or S. More preferably, the heterocycle is a 4-7 membered saturated heterocycle that can be optionally substituted, wherein the cyclizing atoms comprise 1, 2, 3, or 4 heteroatoms selected from N, O, or S; more preferably, the heterocycle is a 5-7 membered (e.g., 5-6 membered) saturated heterocycle that can be optionally substituted, wherein the cyclizing atoms comprise 1, 2, or 3 heteroatoms selected from N, O, or S. The heterocycle may include one or more double or triple bonds and may be a non-aromatic or aromatic ring (e.g., heterocyclic alkyl, heterocyclic alkenyl, or heteroaryl). Examples of heterocycles include, but are not limited to, aziridine, oxadiazine, tetrahydrofuran, pyrrolidine, imidazoline, pyrazolidine, tetrahydropyran, piperidine, morpholine, thiomorpholine, piperazine, preferably pyrrolidine, piperidine, piperazine, and morpholine. The term "heterocycle," "heterocyclic group," or "subheterocyclic group" may be optionally substituted by one or more substituents, as defined below with respect to "optionally substituted."

[0101] As used herein, the term "aryl" refers to a fully carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π-electron system. For example, as used herein, the term "C6-C6" refers to a carbon monocyclic or fused-ring polycyclic aromatic group. 14 "Aryl" means an aromatic group containing 6 to 14 (e.g., 6 to 12) carbon atoms, such as phenyl or naphthyl. Unless otherwise expressly stated in this specification, aryl groups may be optionally substituted.

[0102] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic) aromatic group having a conjugated π-electron system, the ring atoms of which consist of carbon atoms and at least one heteroatom, for example having 5 to 14 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) ring atoms, wherein there are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 carbon atoms and 1, 2, 3, 4, or 5 identical or different heteroatoms independently selected from N, O, S, or S(=O)₂. One or more ring carbon atoms in the heteroaryl group may be replaced by C(=O). The heteroaryl group may be benzofused. Unless otherwise expressly stated in this specification, the heteroaryl group is optionally substituted.

[0103] As used herein, the term "optionally substituted" means that one or more hydrogen atoms attached to an atom or group are independently unsubstituted or independently substituted by one or more (e.g., 1, 2, 3, or 4) substituents. The substituents may be independently selected from, but are not limited to: halogens (e.g., chlorine, bromine, fluorine, or iodine), carboxylic acids (e.g., -C(=O)OH), oxygen (e.g., =O), sulfur (e.g., =S), hydroxyl groups (e.g., -OH), ester groups (e.g., -C(=O)ORiii or -OC(=O)Riii), aldehyde groups (e.g., -C(=O)H), carbonyl groups (e.g., -C(=O)Riii, or represented by C=O), acyl halide groups (e.g., -C(=O)Xi, where Xi is selected from bromine, fluorine, chlorine, or iodine), carbonate groups (e.g., -OC(=O)ORiii), alkoxy groups (e.g., -ORiii), acetal groups (e.g., -C(ORiii)2Riii, where each ORiii is the same or different alkoxy groups), phosphate groups (e.g., P(=O)4), and acetal groups (e.g., P(=O)4). 3- ), mercapto (e.g., -SH), sulfoxide (e.g., -S(=O)Riii), sulfinic acid (e.g., -S(=O)OH), sulfonic acid (e.g., -S(=O)2OH), thioaldehyde (e.g., -C(=S)H), sulfate (e.g., S(=O)4) 2-), sulfonyl (e.g., -S(=O)2Riii), sulfinyl (e.g., -S(=O)Riii), amide (e.g., -C(=O)N(Riii)2 or -N(Riii)C(=O)Riii), azide (e.g., -N3), nitro (e.g., -NO2), cyano (e.g., -CN), isocyano (e.g., -NC), acyloxy (e.g., -OC(=O)Riii), amino (e.g., -NRiii2, -N(Riii)H or -NH2), carbamoyl (e.g., -OC(=O)NRiii2, -OC(=O)N(Riii)H or -OC(=O)NH2), sulfonamide (e.g., -S(=O)2 NRiii2, -S(=O)2NRiiiH, -S(=O)2NH2, -N(Riii)S(=O)2Riii, -N(H)S(=O)2Riii, -N(Riii)S(=O)2H or -N(H)S(=O)2H), alkyl, alkenyl, ynyl, carbocyclic (e.g., cycloalkyl, cycloalkenyl or cycloynyl), heterocyclic (e.g., containing one or more heterocyclic alkyl groups selected from S, N or O heteroatoms, or containing one or more heterocyclic alkenyl groups selected from S, N or O heteroatoms), aryl (e.g., phenyl, or fused-ring group), heteroaryl (e.g., 8-10 membered bicyclic heteroaryl containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur). In any of the foregoing, Riii is hydrogen, alkyl, alkenyl, ynyl, heteroalkyl, heteroalkenyl or heteroynyl as defined herein. In some embodiments, Riii is hydrogen, C1-C1, C2-C3 as defined herein. 12 Alkyl, C2-C 12 alkenyl, C2-C 12 Alkyne group, C1-C 12 Heteroalkyl, C2-C 12 Heterene or C2-C 12 The substituent itself may be further substituted with one or more substituents as defined herein. For example, the C1-C6 alkyl group that is a substituent may be further substituted with one or more substituents as described herein.

[0104] As used herein, the term “halogenated” or “halogenated” is defined as including F, Cl, Br, or I.

[0105] As used herein, "pharmaceutically acceptable salt" refers to an acid addition salt or a base addition salt of the compounds disclosed herein, which retains the biological efficacy and properties of the compounds disclosed herein and is generally not a biologically or otherwise undesirable salt. In many cases, the compounds of the present invention are capable of forming acid salts and / or base salts due to the presence of amino and / or carboxyl groups or similar groups.

[0106] Pharmaceutically acceptable acid addition salts can be formed with inorganic and / or organic acids from the compounds disclosed herein, such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, and phosphoric acid; and such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, camphoric acid, camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, and galactose. Diosmic acid, gentianic acid, glucoheponic acid, gluconic acid, glucuronic acid, glutamic acid, glutamate, 2-oxoglutamate, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucoic 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, dihydroxynaphthalic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanate, p-toluenesulfonic acid, trifluoroacetic acid, and undecenoic acid, etc.

[0107] Pharmaceutically acceptable base addition salts can be formed by reacting inorganic and / or organic bases with the compounds disclosed herein. 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. Salts derived from organic bases include, but are not limited to, primary amines, secondary amines, tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and salts of basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, tannin, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benzylamine, benzathine penicillin, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, and salts of polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0108] The numerical ranges described herein should be understood to encompass boundary values ​​and any and all subranges contained therein. For example, the range “1 to 10” should be understood to include not only the explicitly stated values ​​of 1 and 10, but also any single value within the range of 1 to 10 (e.g., 2, 3, 4, 5, 6, 7, 8, and 9) and subranges (e.g., 1-2, 1.5-2.5, 1-3, 1.5-3.5, 2.5-4, 3-4.5, etc.). This principle also applies to ranges where only one value is used as the minimum or maximum value.

[0109] As used herein, the term "isomer" refers to different compounds having the same molecular formula. "Stereoisomers" are isomers that differ only in the spatial arrangement of their atoms. "Restricted rotational isomers" are stereoisomers obtained by restricted rotation around a single bond. "Enantiomers" are a pair of stereoisomers that are non-overlapping mirror images of each other. A mixture of any proportion of a pair of enantiomers may be called a "racemic" mixture. "Diadiaomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. "Tautomers" refer to the balanced isomeric forms of a compound. The concentration of isomers will depend on the environment in which the compound is found, such as whether the compound is a solid or in an organic or aqueous solution.

[0110] In some embodiments, "stereoisomers" may also include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof.

[0111] Nucleic acids and / or polynucleotides that may be used in this disclosure include a coding region encoding the polypeptide of interest, a 5′-UTR located at the 5′ end of the coding region, and a 3′-UTR located at the 3′ end of the coding region. In some embodiments, the nucleic acid or polynucleotide may also include at least one of a polyadenylated region and a Kozak sequence. In some embodiments, the nucleic acid or polynucleotide (e.g., mRNA) may also include a 5′ cap structure. Any region of the nucleic acid may include one or more alternative nucleosides, such as 5-substituted uridine (e.g., 5-methoxyuridine), 1-substituted pseudouridine (e.g., 1-methyl-pseudouridine or 1-ethyl-pseudouridine), and / or 5-substituted cytidine (e.g., 5-methyl-cytidine).

[0112] The term "5′-UTR" or "5′ untranslated region" can refer to an RNA sequence in mRNA that is upstream of the coding sequence and is not translated into protein. A 5′-UTR in a gene typically begins at the transcription start site and ends at a nucleotide upstream of the translation start codon in the coding sequence. The 5′-UTR can contain elements that control gene expression, such as ribosome binding sites, 5′-terminal oligopyrimidine bundles, and translation initiation signals such as the Kozak sequence. mRNA can be post-transcribed by adding a 5′ cap. Therefore, the 5′-UTR in mature mRNA can also refer to the RNA sequence between the 5′ cap and the start codon. As used herein, the term "3′ untranslated region" or "3′-UTR" can refer to an RNA sequence in mRNA that is downstream of the coding sequence and is not translated into protein. A 3′-UTR in mRNA lies between the stop codon and the poly(A) sequence in the coding sequence, for example, starting at a nucleotide downstream of the stop codon and ending at a nucleotide upstream of the poly(A) sequence. The sequences of the 5′-UTR and / or 3′-UTR may be homologous or heterologous to the sequences of the coding region. The 3′-UTR may contain a 3′-UTR derived from at least one of the following genes: albumin gene, α-globin gene, β-globin gene, tyrosine hydroxylase gene, lipoxygenase gene, and collagen α gene.

[0113] As used herein, the terms "poly(A) region" and "poly(A) sequence" are used interchangeably, and naturally occurring poly(A) sequences typically consist of adenine ribonucleotides. Preferably, a "poly(A) region" refers to a poly(A) sequence containing nucleotides or nucleotide segments other than adenine ribonucleotides. The poly(A) sequence is typically located at the 3′ end of mRNA, such as the 3′ end (downstream) of a 3′-UTR. Poly-A regions can have different lengths. Specifically, in some embodiments, the poly-A region of the nucleic acid molecule of this disclosure is at least 30 nucleotides long; in some embodiments, the poly-A region of the nucleic acid molecule of this disclosure is at least 80 nucleotides long; and in some embodiments, the poly-A region of the nucleic acid molecule of this disclosure is at least 100 nucleotides long.

[0114] As used herein, the term "5′ cap structure" is typically located at the 5′ end of mature mRNA. In some embodiments, the 5′ cap structure is linked to the 5′ end of the mRNA via a 5′-5′-triphosphate bond. The 5′ cap structure is typically formed from modified (e.g., methylated) ribonucleotides, particularly guanine nucleotide derivatives. For example, m7GpppN (cap0, or "cap0") is a cap structure formed by the reaction of the 5′ phosphate group of hnRNA with the 5′ phosphate group of m7GTP under the action of guanylate transferase to form a 5′,5′-phosphodiester bond, where N is the terminal 5′ nucleotide of the nucleic acid carrying the 5′ cap structure. In some implementations, the 5′ cap structure includes, but is not limited to, cap 0, cap 1 (a cap structure formed by further methylation of the 2'-OH of the first nucleotide glycosyl group of hnRNA on the basis of cap 0, or "cap1"), cap 2 (a cap structure formed by further methylation of the 2'-OH of the second nucleotide glycosyl group of hnRNA on the basis of cap 1, or "cap2"), cap 4, cap 0 analogue, cap 1 analogue, cap 2 analogue, or cap 4 analogue.

[0115] amino lipid compounds

[0116] This disclosure provides amino lipid compounds as shown in formula (I), formula (II) or formula (III):

[0117] Or its pharmaceutically acceptable salt, or its stereoisomer,

[0118] in,

[0119] X1 and X2 are independently selected from O or S;

[0120] R1 is Where R ’ H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 heteroalkyl, or optionally substituted C2-C6 heteroalkenyl; R ” H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C2-C6 heteroalkenyl, or

[0121] X3 and X4 are independently selected from O or S;

[0122] R M H, halogen, -OH, -R * -N(R) * )2, -CN, -N3, -C(=O)OH, -C(=O)OR * -OC(=O)R* -OR * -SR * -S(=O)R * -S(=O)OR * -S(=O)2OR * -NO2, -S(=O)2N(R) * )2、-N(R * )S(=O)2R * -NH(CH2) t1 N(R * 2、-NH(CH2) p1 O(CH2) q1 N(R * 2、-NH(CH2) s1 OR * -N((CH2) s1 OR * )2、-N(R * -Carbon ring, -N(R) * - Heterocyclic rings, -N(R) * -Aryl, -N(R) * )-Heteroaryl, -N(R * (CH2) t1 -Carbon ring, -N(R) * (CH2) t1 -heterocyclic, -N(R) * (CH2) t1 -Aryl, -N(R) * (CH2) t1 - Heteroaryl, carbocyclic, heterocyclic, aryl or heteroaryl;

[0123] Each R * It is independently H, C1-C3 alkyl, or C2-C3 alkenyl;

[0124] Each t1 is independently 1, 2, 3, 4 or 5;

[0125] p1 is 1, 2, 3, 4 or 5;

[0126] q1 is 1, 2, 3, 4, or 5;

[0127] Each s1 is independently 1, 2, 3, 4 or 5;

[0128] R N It is H or C1-C3 alkyl;

[0129] r is 0 or 1;

[0130] n is any integer from 0 to 10;

[0131] L3 is a bond, an optional substituted C1-C6 alkylene group, or an optional substituted C1-C6 heteroalkylene group;

[0132] R2 is H, a C1-C6 hydrocarbon group, or a C1-C6 heterohydrocarbon group;

[0133] L1 and L2 are independently selected from bonded, optionally substituted C1-C3 alkylene or optionally substituted C2-C3 alkenyl groups;

[0134] A1 and A2 are independently selected from C1 to C2. 10 Alkylene or C1-C 10 Heteroalkyl;

[0135] A3 and A4 are independently selected from C1-C6 alkylene, C1-C6 heteroalkylene, or bonded;

[0136] R3 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5);

[0137] R4 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6);

[0138] Each M1 is independently C1~C 24 Hydroxyl group;

[0139] Each M2 is independently C1 to C 12 Hydroxyl group;

[0140] Each M a Independently C1~C 24 hydrocarbon group;

[0141] Each M b Independently C1~C 15 hydrocarbon group;

[0142] Z1 and Z2 are independently selected from -C(=O)O- or -OC(=O)-;

[0143] Each Ra Independently, it is an H or C1-C8 hydrocarbon group;

[0144] Each R5 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ;

[0145] Each R6 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ;

[0146] Each M3 is independently C3~C 12 Hydroxyl group;

[0147] Each M4 is independently a C1–C8 hydrocarbon group;

[0148] Each M c Independently C3~C 12 hydrocarbon group;

[0149] Each M d Independently composed of C1–C8 hydrocarbon groups;

[0150] Each Y is an independent 3-7 member carbon ring.

[0151] The aminolipid compounds represented by formula (I) or formula (II) may include one or more of the following features when applicable.

[0152] In some implementations, X1 and X2 are 0.

[0153] In some implementations, Z1 and Z2 are -C(=O)O-.

[0154] In some implementations, R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is -C(R a (OR6)2、-C(R) a (SR6)2 or -C(R) a )(SR6)(OR6).

[0155] In some implementations, R3 is C1 to C2. 24 hydrocarbon group or -C(R) a (OR5)2, R4 is -C(R a )(OR6)2.

[0156] In some implementations, R3 is C1 to C2. 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0157] In some implementations, R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2.

[0158] In some implementations, R3 is -C(R a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is C1~C 24 hydrocarbon group, -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a )(SR6)(OR6).

[0159] In some implementations, R3 is -C(R a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a )(OR6)2.

[0160] In some implementations, R3 is -CH(OR5)2, and R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0161] In some implementations, R3 is -CH(OR5)2, and R4 is C1 to C2. 24 Hydrocarbon group.

[0162] In some implementations, R3 is -CH(OR5)2 and R4 is -CH(OR6)2.

[0163] In some implementations, X1 and X2 are 0, and R3 is C1 to C2. 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0164] In some implementations, X1 and X2 are 0, and R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2.

[0165] In some implementations, X1 and X2 are O, R3 is -CH(OR5)2, and R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0166] In some implementations, X1 and X2 are O, R3 is -CH(OR5)2, and R4 is C1 to C2. 24 Hydrocarbon group.

[0167] In some implementations, X1 and X2 are O, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0168] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 The hydrocarbon group is -CH(OR5)2, and R4 is -CH(OR6)2.

[0169] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2.

[0170] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group or -CH(OR6)2.

[0171] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group.

[0172] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0173] The aminolipid compounds represented by formula (III) include one or more of the following features when applicable.

[0174] In some implementations, X1 and X2 are 0.

[0175] In some implementations, Z1 and Z2 are -C(=O)O-.

[0176] In some implementations, R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is C1~C 24 hydrocarbon group, -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a )(SR6)(OR6).

[0177] In some implementations, R3 is C1 to C2. 24 Hydrocarbon group, R4 is C1 to C2. 24 Hydrocarbon group.

[0178] In some implementations, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a )(OR6)2.

[0179] In some implementations, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0180] In some implementations, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2.

[0181] In some implementations, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0182] In some implementations, R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0183] In some implementations, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2 to C2. 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0184] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a(OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a )(OR6)2.

[0185] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2.

[0186] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2.

[0187] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2.

[0188] In some implementations, X1 and X2 are 0, and R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0189] In some implementations, X1 and X2 are O, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0190] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0191] In some implementations, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkylene. In some embodiments, A1 or A2 is a C4-C9 alkylene. In some embodiments, A1 and A2 are independently selected from C4-C9 alkylene, for example, C5-C8 alkylene.

[0192] The aminolipid compounds represented by formula (I), formula (II) or formula (III) may include one or more of the following features when applicable.

[0193] In some implementations, for C1 to C3 as defined by R3 24 Each hydrocarbon group is independently C1 to C2. 24 Alkyl or C2-C 24 Alkenyl group.

[0194] In some implementations, for C1 to C3 as defined by R3 24 Each hydrocarbon group is independently C3 to C4. 20 Alkyl or C3-C 20 Alkenyl group.

[0195] In some implementations, for C1 to C3 as defined by R3 24 Each hydrocarbon group is independently C5 to C6. 18 alkyl.

[0196] In some implementations, for C1 to C3 as defined by R3 24 Each hydrocarbon group is independently a straight-chain C3-C10 group. 15 Alkyl groups, such as straight-chain C5-C6 groups. 12 Alkyl or straight-chain C6-C 10 alkyl.

[0197] In some implementations, for C1 to C3 as defined by R3 24 Each hydrocarbon group is independently branched C8-C. 24 Alkyl groups, such as branched C8-C6 groups 20 Alkyl or branched C 10 ~C 18 alkyl.

[0198] In some implementations, for C1 to C4 as defined by R4 24 Each hydrocarbon group is independently C1 to C2. 24 Alkyl or C2-C 24 Alkenyl group.

[0199] In some implementations, for C1 to C4 as defined by R4 24 Each hydrocarbon group is independently C3 to C4. 20 Alkyl or C3-C 20 Alkenyl group.

[0200] In some implementations, for C1 to C4 as defined by R4 24 Each hydrocarbon group is independently C5-C6. 18 alkyl.

[0201] In some implementations, for C1 to C4 as defined by R4 24 Each hydrocarbon group is independently a straight-chain C3-C10 group. 15 Alkyl groups, such as straight-chain C5-C6 groups. 12 Alkyl or straight-chain C6-C 10 alkyl.

[0202] In some implementations, for C1 to C4 as defined by R4 24 Each hydrocarbon group is independently branched C8-C. 24 Alkyl groups, such as branched C8-C6 alkyl groups 20 Alkyl or branched C 10 ~C 18 alkyl.

[0203] In some embodiments, the aminolipid compound represented by formula (I) has a structure as shown in formula (IV):

[0204] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in this paper for equation (I).

[0205] In some embodiments, the aminolipid compound represented by formula (II) has a structure as shown in formula (V):

[0206] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in equation (II) in this paper.

[0207] In some embodiments, the aminolipid compound represented by formula (III) has a structure as shown in formula (VI):

[0208] Among them, R1, R2, L1, L2, A1, A2, A3, A4, R5 and R6 are as defined in this paper for equation (III).

[0209] The aminolipid compounds represented by formula (I), formula (II), formula (III), formula (IV), formula (V) or formula (VI) may include one or more of the following features when applicable.

[0210] In some embodiments, L1 and L2 are independently selected from bonded or optionally substituted C1-C3 alkylene groups, and the ring containing L1 and L2 is a 4-7 membered heterocycle. In some embodiments, L1 and L2 are independently selected from bonded or unsubstituted C1-C3 alkylene groups. In some embodiments, the heterocycle is a 5-7 membered heterocycle, such as a 5-membered or 6-membered heterocycle.

[0211] As used herein, for aminolipid compounds represented by formula (I), formula (II) or formula (III), "the ring containing L1 and L2" refers to the heterocycle formed by L1, L2, X1 connected to L1, X2 connected to L2, the carbon atoms connecting X1 and X2, and the carbon atoms connecting L1 and L2.

[0212] As used herein, for aminolipid compounds represented by formula (IV), (V) or (VI), "the ring containing L1 and L2" refers to the heterocycle formed by L1, L2, the O atom connected to L1, the O atom connected to L2, the carbon atom connecting the two O atoms, and the carbon atom connecting L1 and L2.

[0213] In some implementations, L1 and L2 are not both keys.

[0214] In some embodiments, L1 and L2 are not simultaneously unsubstituted C1 alkylene groups. For example, L1 is a bonded, unsubstituted C1 or C2 alkylene group, and L2 is a bonded group. As another example, L1 is a bonded group, and L2 is a bonded, unsubstituted C1 or C2 alkylene group.

[0215] In some embodiments, L1 is a bond and L2 is an unsubstituted C1 or C2 alkylene, for example, an unsubstituted C1 alkylene.

[0216] In some embodiments, L1 is an unsubstituted C1 or C2 alkylene, for example an unsubstituted C1 alkylene, and L2 is a bond.

[0217] In other implementations, L1 and L2 are both keys.

[0218] In some implementation schemes, R ’ H, an optionally substituted C1-C6 alkyl group, or an optionally substituted C2-C6 alkenyl group; R ” H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl or

[0219] In some implementation schemes, R ’and R ” Independently selected from optionally substituted C1-C6 alkyl or optionally substituted C2-C6 alkenyl groups. For example, R ’ and R ” Independently selected from optional substituted C1 to C3 alkyl groups.

[0220] In some implementation schemes, R ’ and R ” Independently selected from unsubstituted C1-C3 alkyl groups. For example, R ’ and R ” It is independently selected from unsubstituted C1 to C2 alkyl groups.

[0221] In some implementations, X3 and X4 are O, R M -N(R) * )2, r is 1. In some implementations, each R * Independently, it is an H or C1-C3 alkyl group. In some embodiments, one of the R groups is... * For H. In some implementations, an R * For H, another R * It is a C1 to C3 alkyl group.

[0222] In some implementation schemes, R ’ R is an optionally substituted C1-C3 alkyl group. ” for R * It is a C1-C3 alkyl group. In some embodiments, R ’ It is an unsubstituted C1 to C3 alkyl group, such as R ’ It is methyl or ethyl. In some embodiments, R * It can be methyl or ethyl.

[0223] In some implementation schemes, R N For H.

[0224] In some implementations, n is any integer from 1 to 5. For example, n is 2, 3, or 4.

[0225] In some embodiments, L3 is a bond, an optionally substituted C1-C6 alkylene group, an optionally substituted C2-C6 alkenyl group, an optionally substituted C1-C6 heteroalkylene group, or an optionally substituted C2-C6 heteroalkylene group.

[0226] In some embodiments, L3 is a bond, an optionally substituted C1-C6 alkylene group, or an optionally substituted C1-C6 heteroalkylene group.

[0227] In some embodiments, L3 is a bonded, optionally substituted C1-C6 alkylene or optionally substituted C2-C6 alkenylene. In some embodiments, L3 is a bonded or optionally substituted C1-C6 alkylene. In some embodiments, L3 is an unsubstituted C1-C4 alkylene, for example, L3 is an unsubstituted C1-C3 alkylene.

[0228] In some embodiments, R2 is H, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 heteroalkyl, or C2-C6 heteroalkenyl.

[0229] In some embodiments, R2 is H, C1-C6 alkyl, or C1-C6 heteroalkyl.

[0230] In some embodiments, R2 is H, C1-C6 alkyl, or C2-C6 alkenyl.

[0231] In some embodiments, R2 is H or a C1-C6 alkyl group.

[0232] In some implementations, R2 is H.

[0233] In other embodiments, R2 is a C1-C3 alkyl group. For example, R2 is a C1-C2 alkyl group.

[0234] In some implementations, R3 is C1 to C2. 24 Alkyl, C2-C 24 Alkenyl, C1-C containing O or S 24 Heteroalkyl groups, C2-C groups containing O or S 24 Heterene, -M1Y, -YM a -M2YM b -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a )(SR5)(OR5).

[0235] In some implementations, R3 is C1 to C2. 24 Alkyl, C2-C 24 Alkenyl, C1-C containing O or S 24 Heteroalkyl groups or C2-C groups containing O or S 24 Heterene group.

[0236] In some implementations, R3 is C1 to C2. 24 Alkyl or C2-C 24 Alkenyl group.

[0237] In some implementations, R3 is C3 to C 20 Alkyl or C3-C 20 Alkenyl group.

[0238] In some implementations, R3 is C5 to C 18 alkyl.

[0239] In some implementations, R3 is a straight-chain C3-C13. 15 Alkyl groups, such as straight-chain C5-C6 groups. 12 Alkyl or straight-chain C6-C 10 alkyl.

[0240] In some implementations, R3 is a branched C8-C 24 Alkyl groups, such as branched C8-C6 alkyl groups 20 Alkyl or branched C 10 ~C 18 alkyl.

[0241] In some implementations, R3 is C3-C containing O or S. 20 Heteroalkyl groups.

[0242] In some implementations, R3 is a C5-C5 compound containing O or S. 18 Heteroalkyl groups.

[0243] In some implementations, R4 is C1 to C2. 24 Alkyl, C2-C 24 Alkenyl, C1-C containing O or S 24 Heteroalkyl groups, C2-C groups containing O or S 24 Heterene, -M1Y, -YM a -M2YM b -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a )(SR6)(OR6).

[0244] In some implementations, R4 is C1 to C2. 24 Alkyl, C2-C 24 Alkenyl, C1-C containing O or S 24 Heteroalkyl groups or C2-C groups containing O or S 24 Heterene group.

[0245] In some implementations, R4 is C1 to C2. 24 Alkyl or C2-C 24 Alkenyl group.

[0246] In some implementations, R4 is C3 to C4. 20 Alkyl or C3-C 20 Alkenyl group.

[0247] In some implementations, R4 is C5 to C 18 alkyl.

[0248] In some implementations, R4 is a straight-chain C3-C4. 15 Alkyl groups, such as straight-chain C5-C6 groups. 12 Alkyl or straight-chain C6-C 10 alkyl.

[0249] In some implementations, R4 is a branched C8 to C96 branch. 24 Alkyl groups, such as branched C8-C6 alkyl groups 20 Alkyl or branched C 10 ~C 18 alkyl.

[0250] In some implementations, R4 is C3-C containing O or S. 20 Heteroalkyl groups.

[0251] In some implementations, R4 is a C5-C group containing O or S. 18 Heteroalkyl groups.

[0252] In some implementations, R3 is -M1Y or -YM. a or -M2YM b .

[0253] In some implementations, R4 is -M1Y or -YM. a or -M2YM b .

[0254] In some implementations, each M1 is independently C1 to C2. 24 Alkylene or C2-C 24 Alkenyl group.

[0255] In some implementations, each M1 is independently C1 to C2. 24 Alkylene.

[0256] In some implementation schemes, each M1 is independently C5 to C 18 Alkylene, for example, independently C5-C6 15 Alkylene, C8-C 12 Alkylene, C6-C 13 Alkylene.

[0257] In some implementation schemes, each M a Independently C1~C 24 Alkyl or C2-C 24 Alkenyl group.

[0258] In some implementation schemes, each M a Independently C1~C 24 alkyl.

[0259] In some implementation schemes, each M a Independently C5~C 18 Alkyl groups, for example, independently C5-C6. 15 Alkyl, C8-C 12 Alkyl, C6-C 13 alkyl.

[0260] In some implementation schemes, each M2 is independently C1 to C2. 12 Alkylene or C2-C 12 Alkenyl group.

[0261] In some implementation schemes, each M2 is independently C1 to C2. 12 Alkylene.

[0262] In some implementation schemes, each M2 is independently C3 to C 10 Alkylenes, for example, independently C4-C9 alkylenes or C5-C8 alkylenes.

[0263] In some implementation schemes, each M b Independently C1~C 15 Alkyl or C2-C 15 Alkenyl group.

[0264] In some implementation schemes, each M b Independently C1~C 15 alkyl.

[0265] In some implementation schemes, each M b Independently C3~C 12 Alkyl groups, for example, independently C4-C5. 10 Alkyl, C5-C8 alkyl.

[0266] In some implementations, in each -M2YM b In the middle, M2 and M b The total number of carbons in the main chain is independently 2 to 20, for example 4 to 18, 6 to 12, 8 to 10.

[0267] In some implementations, each R5 is independently C3 to C5. 12 Alkyl, C3-C 12 alkenyl, C3~C 12 Heteroalkyl, C3-C 12 Heterene, -M3Y, -YM c or -M4YM d .

[0268] In some implementations, each R5 is independently C3 to C5. 12 Alkyl, C3-C 12alkenyl, C3~C 12 Heteroalkyl, or C3-C 12 Heterene group.

[0269] In some implementations, each R5 is independently C3 to C5. 12 Alkyl or C3-C 12 Alkenyl group.

[0270] In some implementations, each R5 is independently C3 to C5. 12 Alkyl group. In some embodiments, each R5 is independently a C3 to C9 alkyl group. In some embodiments, each R5 is independently a C5 to C8 alkyl group.

[0271] In some embodiments, each R5 is independently a straight-chain C5-C8 alkyl group. In some embodiments, each R5 is independently a branched C5-C8 alkyl group.

[0272] In some implementations, R5 is C3 to C5. 12 Alkyl or C3-C 12 Alkenyl group.

[0273] In some implementations, R5 is C3 to C5. 12 Alkyl group. In some embodiments, R5 is a C3 to C9 alkyl group. In some embodiments, R5 is a C5 to C8 alkyl group.

[0274] In some embodiments, R5 is a straight-chain C5-C8 alkyl group. In some embodiments, R5 is a branched C5-C8 alkyl group.

[0275] In some implementations, each R5 is independently C3 to C5. 12 Heteroalkyl. In some embodiments, each R5 is independently a C3 to C9 heteroalkyl.

[0276] In some implementations, each R6 is independently C3 to C6. 12 Alkyl, C3-C 12 alkenyl, C3~C 12 Heteroalkyl, C3-C 12 Heterene, -M3Y, -YM c or -M4YM d .

[0277] In some implementations, each R6 is independently C3 to C6. 12 Alkyl, C3-C 12 alkenyl, C3~C 12 Heteroalkyl, or C3-C 12 Heterene group.

[0278] In some implementations, each R6 is independently C3 to C6. 12Alkyl or C3-C 12 Alkenyl group.

[0279] In some implementations, each R6 is independently C3 to C6. 12 Alkyl group. In some embodiments, each R6 is independently a C3 to C9 alkyl group. In some embodiments, each R6 is independently a C5 to C8 alkyl group.

[0280] In some embodiments, each R6 is independently a straight-chain C5-C8 alkyl group. In some embodiments, each R6 is independently a branched C5-C8 alkyl group.

[0281] In some implementations, R6 is C3 to C6. 12 Alkyl or C3-C 12 Alkenyl group.

[0282] In some implementations, R6 is C3 to C6. 12 Alkyl group. In some embodiments, R6 is a C3-C9 alkyl group. In some embodiments, R6 is a C5-C8 alkyl group.

[0283] In some embodiments, R6 is a straight-chain C5-C8 alkyl group. In some embodiments, R6 is a branched C5-C8 alkyl group.

[0284] In some implementations, each R6 is independently C3 to C6. 12 Heteroalkyl. In some embodiments, each R6 is independently a C3 to C9 heteroalkyl group.

[0285] In some implementations, each R5 is independently -M3Y, -YM c or -M4YM d .

[0286] In some implementations, each R6 is independently -M3Y, -YM c or -M4YM d .

[0287] In some implementation schemes, each M3 is independently C3 to C 12 Alkylene or C3-C 12 Alkenyl group.

[0288] In some implementation schemes, each M3 is independently C3 to C 12 Alkylene.

[0289] In some implementation schemes, each M3 is independently C3 to C 10 Alkylenes, for example, independently C4-C9 alkylenes or C5-C8 alkylenes.

[0290] In some implementation schemes, each M cIndependently C3~C 12 Alkyl or C3-C 12 Alkenyl group.

[0291] In some implementation schemes, each M c Independently C3~C 12 alkyl.

[0292] In some implementation schemes, each M c Independently C3~C 10 Alkyl groups, for example, independently C4-C9 alkyl groups or C5-C8 alkyl groups.

[0293] In some embodiments, each M4 is independently a C1-C8 alkylene group or a C2-C8 alkenyl group.

[0294] In some implementations, each M4 is independently a C1-C8 alkylene group.

[0295] In some embodiments, each M4 is independently a C2-C6 alkylene group, for example, independently a C2-C5 alkylene group or a C3-C4 alkylene group.

[0296] In some implementation schemes, each M d It is independently a C1-C8 alkyl or C2-C8 alkenyl.

[0297] In some implementation schemes, each M d It is independently a C1 to C8 alkyl group.

[0298] In some implementation schemes, each M d It is independently a C2 to C6 alkyl group, for example, independently a C2 to C5 alkyl group or a C3 to C4 alkyl group.

[0299] In some implementations, in each -M4YM d In the middle, M4 and M d The total number of carbons in the main chain is independently 2 to 13, for example 4 to 10, 6 to 8.

[0300] In some implementations, each Y is independently a 4-6 membered carbon ring, for example a 4-6 membered monocyclic carbon ring.

[0301] In some implementations, each Y is independently a 4-6 saturated monocyclic carbon ring.

[0302] In some implementation schemes, each R a It is independently H or C1 to C8 alkyl.

[0303] In some implementation schemes, each R a It is independently H or C1 to C5 alkyl.

[0304] In some implementation schemes, each Ra It is independently H or C1-C2 alkyl.

[0305] In some implementation schemes, R a For H.

[0306] In some implementations, A1 and A2 are independently C1 to C2. 10 Alkylene. For example, A1 and A2 are independently C3-C4. 10 Alkylene or C4-C9 alkylene.

[0307] In some embodiments, A1 and A2 are independently C5 to C9 alkylene groups. For example, A1 and A2 are independently C5 to C8 alkylene groups.

[0308] In some implementations, A1 is a C5-C8 alkylene group.

[0309] In some implementations, A2 is a C5-C8 alkylene group.

[0310] In some embodiments, A3 and A4 are independently C1-C6 alkylene groups. For example, A3 and A4 are independently C1-C6 alkylene groups; or, A3 and A4 are independently C1-C6 alkylene groups. In some embodiments, A3 and A4 are independently C1-C4 alkylene groups. In some embodiments, A3 and A4 are independently C2-C4 alkylene groups.

[0311] In some implementations, A3 and A4 are C3 alkylene groups.

[0312] In some implementations, A3 and A4 are keys.

[0313] In some implementations, A3 is the key.

[0314] In some implementations, A4 is the key.

[0315] In some implementations, the hydrocarbon group is alkyl, alkenyl, or ynyl.

[0316] In some implementations, the heteroalkyl group is a heteroalkyl, heteroalkenyl, or heteroyne group.

[0317] In some embodiments, the heteroatom in the heteroalkyl, heteroalkyl, heteroalkenyl, or heteroynyl group as defined herein is one or more of N, O, or S.

[0318] In some embodiments, the aminolipid compound is selected from one of the structures shown in Table 1:

[0319] Table 1

[0320] The aminolipid compounds disclosed herein contain long, nonpolar residues, all of which are hydrophobic. Furthermore, the aminolipid compounds also possess an amino group, thus exhibiting hydrophilic properties. Utilizing this amphiphilic characteristic, the aminolipid compounds of this disclosure can be used to prepare nanoparticles, such as lipid nanoparticles (LNPs), lipid bilayers, micelles, and liposomes. The prepared nanoparticles, especially lipid nanoparticles, can serve as delivery carriers for delivering active ingredients such as nucleic acids, thereby enhancing the efficacy of gene therapy.

[0321] lipid nanoparticles

[0322] This disclosure provides lipid nanoparticles, such as empty lipid nanoparticles (empty LNPs) or loaded lipid nanoparticles (loaded LNPs).

[0323] As used herein, empty lipid nanoparticles refer to nanoparticles containing only lipid components; loaded lipid nanoparticles refer to nanoparticles containing not only lipid components but also other components (such as active ingredients).

[0324] On the one hand, this disclosure provides empty lipid nanoparticles containing the amino lipid compounds of this disclosure.

[0325] In some implementations, the empty lipid nanoparticles also contain one or more of the following: auxiliary lipids, structural lipids, and PEG-lipids (polyethylene glycol-lipids).

[0326] In some implementations, the empty lipid nanoparticles also contain auxiliary lipids, structural lipids, and PEG-lipids.

[0327] In some embodiments, the empty lipid nanoparticles contain the following amounts (molar percentages) of aminolipid compounds, based on the total amount of aminolipid compounds, auxiliary lipids, structural lipids, and PEG-lipids: about 30.0%-60.0%, for example, about 32.0%-58.0%, 35.0%-55.0%, 37.0%-53.0%, 40.0%-50.0%, 42.0%-48.0%, 30.0%-55.0%, 30.0%-50.0%, 30.0%-45.0%, 30.0%-40.0%, 30.0%-35.0%, 35.0%-60.0%, 35.0%-50.0%. 35.0%-45.0%, 40.0%-60.0%, 40.0%-55.0%, 40.0%-45.0%, 45.0%-60.0%, 45.0%-55.0%, 45.0%-50.0%, 50.0%-60.0%, 30.0%-32.0%, 32.0%-35.0%, 35.0%-40.0%, 40.0%-42.0%, 42.0%-45.0%, 45.0%-46.3%, 46.3%-48.0%, 48.0%-49.5%, 49.5%-50.0%, 50.0%-55.0%, or 55.0%-60.0%.

[0328] In some embodiments, the empty lipid nanoparticles contain the following amounts (molar percentages) of auxiliary lipids, based on the total amount of aminolipid compounds, assist lipids, structural lipids, and PEG-lipids: about 0%-30.0%, for example about 2.0%-28.0%, 5.0%-25.0%, 8.0%-23.0%, 10.0%-20.0%, 12.0%-18.0%, 0%-25.0%, 0.0%-20.0%, 0.0%-15.0%, 0.0%-10.0%, 5.0%-30.0%, 5.0%-25.0%, 5.0%-20.0%, 5.0%-15.0%, 5.0%-12.0%, 5.0%-10.0%, 8.0%-30.0%, 8.0%-25.0%, 8%- 20.0%, 8.0%-15.0%, 8.0%-12.0%, 8.0%-10.0%, 10.0%-30.0%, 10.0%-25.0%, 10.0%-23.0%, 10.0%-15.0%, 15.0%-30.0%, 15.0%-25.0%, 15%-20.0%, 20.0%-30.0%, 0 %-5.0%, 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%, 20.0%-25.0%, or 25.0%-30.0%.

[0329] In some embodiments, the empty lipid nanoparticles contain the following amounts (molar percentages) of structural lipids, based on the total amount of aminolipid compounds, auxiliary lipids, structural lipids, and PEG-lipids: approximately 18.5%–48.5%, for example, approximately 20.0%–45.0%, 25.0%–40.5%, 30.5%–35.5%, 18.5%–46.5%, 18.5%–45.0%, 18.5%–40.0%, 18.5%–35.0%. 18.0%-30.5%, 18.5%-25.0%, 18.0%-20.5%, 20.0%-48.5%, 20.0%-45.5%, 20.0%-40.5%, 20.0%-35.5%, 20.0%-30.0%, 20.0%-25.5%, 25.0%-48.5%, 25.0%-44.5%, 25.0%-42.5%, 30.0%-48.5%, 30.3%-43 0.5%, 30.0%-41.0%, 30.0%-36.0%, 35.0%-48.5%, 35.0%-45.5%, 35.0%-42.0%, 35.0%-40.5%, 40.0%-48.5%, 40.0%-45.5%, 18.5%-22.5%, 22.5%-23.5%, 23.5%-28.5%, 28.5%-33.5%, 33.5%-35.0%, 35.0% 36.5%, 36.5%-38.0%, 38.0%-38.5%, 38.5%-39.0%, 39.0%-39.5%, 39.5%-40.5%, 40.5%-41.5%, 41.5%-42.5%, 42.5%-42.7%, 42.7%-43.0%, 43.0%-43.5%, 43.5%-45.0%, 45.0%-46.5%, or 46.5%-48.5%.

[0330] In some embodiments, the empty lipid nanoparticles contain the following amounts (molar percentages) of PEG-lipids, based on the total amount of aminolipid compounds, auxiliary lipids, structural lipids, and PEG-lipids: about 0%-10.0%, for example about 0.5%-9.5%, 1.0%-9.0%, 2.0%-8.0%, 3.0%-7.0%, 4.0%-6.0%, 0%-9.0%, 0%-8.0%, 0%-6.0%, 0%-4.0%, and 0%-2%. 0%, 0.5%-10.0%, 0.5%-9.0%, 0.5%-7.5%, 0.5%-6.0%, 0.5%-4.5%, 0.5%-2.0%, 1.0%-10.0%, 1.0%-7.5%, 1.0%-6.0%, 1.0%-4.5%, 1.0%-2.0%, 2.0%-10.0%, 2.0%-9.0%, 2.0%-7.0%, 2.0%-6.0%, 2.0%-4.0%, 4 0%-10.0%, 4.0%-9.0%, 4.0%-8.0%, 5.0%-10.0%, 5.0%-7.5%, 5.0%-6.0%, 6.0%-10.0%, 6.0%-9.0%, 6.0%-8.0%, 7.0%-10.0%, 7.0%-8.0%, 8.0%-10.0%, 0%-0.5%, 0.5%-1.0%, 1.0%-1.5%, 1.5%-1.6%, 1.6% -2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, 4.5%-5.0%, 5.0%-5.5%, 5.5%-6.0%, 6.0%-6.5%, 6.5%-7.0%, 7.0%-7.5%, 7.5%-8.0%, 8.0%-8.5%, 8.5%-9.0%, 9.0%-9.5%, or 9.5%-10.0%.

[0331] In some embodiments, the cofactor lipid is a phospholipid. Phospholipids are typically semi-synthetic, but can also be of natural origin or chemically modified. Phospholipids include, but are not limited to, DSPC (distearylphosphatidylcholine), DOPE (dioleoylphosphatidylethanolamine), DOPC (dioleoyllecithin), DOPS (dioleoylphosphatidylserine), DSPG (1,2-octacosanyl-sn-glycerol-3-phosphate-(1'-rac-glycerol)), DPPG (dispalmitoylphosphatidylglycerol), DPPC (dispalmitoylphosphatidylcholine), DGTS (1,2-dispalmitoyl-sn-glycerol-3-O-4'-(N,N,N-trimethyl)homoserine), lysophospholipids, etc. In some embodiments, the cofactor lipid is selected from one or more of DSPC, DOPE, DOPC, and DOPS. In some embodiments, the cofactor lipid is DSPC and / or DOPE.

[0332] In some embodiments, the structural lipid is a sterol, including but not limited to cholesterol, cholesterol esters, sterol hormones, sterol vitamins, bile acids, cholesterol, ergosterol, β-sitosterol, and oxidized cholesterol derivatives. In some embodiments, the structural lipid is selected from at least one of cholesterol, cholesterol esters, sterol hormones, sterol vitamins, and bile acids. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid is high-purity cholesterol, particularly injectable high-purity cholesterol, such as CHO-HP (produced by AVT).

[0333] As used herein, the term PEG-lipid (polyethylene glycol-lipid) is a conjugate of polyethylene glycol and a lipid structure. In some embodiments, the PEG-lipid is selected from at least one of PEG-DMG (or DMG-PEG) and PEG-distearate phosphatidylethanolamine (PEG-DSPE), such as PEG-DMG, wherein PEG-DMG is a polyethylene glycol (PEG) derivative of glyceryl 1,2-dimyristate. In some embodiments, the average molecular weight of PEG is about 2000 to 5000. In some embodiments, the average molecular weight of PEG is about 2000.

[0334] In some implementations, the auxiliary lipid is DOPE and the structural lipid is CHO-HP.

[0335] In some implementations, the auxiliary lipid is DSPC and the structural lipid is CHO-HP.

[0336] On the other hand, this disclosure provides loaded lipid nanoparticles comprising empty lipid nanoparticles as described in any of the above embodiments and an active ingredient.

[0337] In some implementations, the active ingredient is a pharmaceutical active ingredient, which is a therapeutic and / or preventative agent.

[0338] As used herein, the terms “therapeutic agent” or “preventive agent” mean any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or preventive effect and / or causes the desired biological and / or pharmacological effect.

[0339] In some implementations, the active pharmaceutical ingredient is a bioactive ingredient, which is a substance that has a biological effect when introduced into cells or a host, for example, by stimulating an immune or inflammatory response, by exerting enzymatic activity, or by supplementing mutations. Bioactive ingredients include, but are not limited to, nucleic acids, proteins, peptides, antibodies, small molecules, and mixtures thereof.

[0340] In some implementations, the bioactive ingredient is nucleic acid.

[0341] In some implementations, the nucleic acid is selected from one or more of RNA, antisense oligonucleotides, and DNA.

[0342] In some implementations, the RNA is selected from messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single-stranded guide RNA (sgRNA), Cas9 mRNA, or a mixture thereof.

[0343] In some embodiments, messenger RNA (mRNA) encodes a polypeptide and / or protein of interest. This includes any polypeptide that is naturally or non-naturally present or otherwise modified. In some embodiments, the polypeptide and / or protein encoded by the mRNA may have therapeutic and / or preventative effects when expressed in cells.

[0344] In some embodiments, the RNA is a siRNA, which can selectively reduce or downregulate the expression of a gene of interest. For example, the choice of siRNA can enable the silencing of genes associated with a specific disease, condition, or ailment after administration of lipid nanoparticles comprising the siRNA to a subject in need. 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.

[0345] In some implementations, the RNA is sgRNA and / or cas9 mRNA. sgRNA and / or cas9 mRNA can be used as gene editing tools. For example, the sgRNA-cas9 complex can affect the mRNA translation of cellular genes.

[0346] In some implementations, the RNA is shRNA or its encoding vector or plasmid. shRNA can be generated inside the target cell after the appropriate construct is delivered to the nucleus. The constructs and mechanisms associated with shRNA are well known in the relevant field.

[0347] In some implementations, the DNA is a plasmid.

[0348] In other embodiments, the bioactive ingredient is an antitumor agent, antibiotic, immunomodulator, anti-inflammatory agent, agent acting on the central nervous system, polypeptide, protein, antigen, or mixture thereof.

[0349] In other embodiments of this disclosure, the active ingredient may also be a transfection reagent, a detection reagent, etc.

[0350] Pharmaceutical Composition

[0351] In another respect, this disclosure provides pharmaceutical compositions comprising the above-described loaded lipid nanoparticles and pharmaceutically acceptable carriers, diluents or excipients.

[0352] In some implementations, the pharmaceutical composition also contains a buffer solution.

[0353] In some implementations, the buffer is selected from phosphate buffer and / or Tris buffer. For example, the buffer is phosphate buffer.

[0354] In some implementations, the concentration of the buffer solution is approximately 5 mmol / L to 30 mmol / L. For example, the concentration of the buffer solution is approximately 10 mmol / L.

[0355] In some implementations, the buffer solution has a pH of approximately 6-8. For example, the buffer solution has a pH of approximately 7-8. Yet another example is a buffer solution with a pH of approximately 7-7.5.

[0356] In some embodiments, the pharmaceutical composition also contains a cryoprotectant.

[0357] In some embodiments, the cryoprotectant is selected from sucrose and / or trehalose. For example, the cryoprotectant is sucrose.

[0358] In some implementations, the concentration of the cryoprotectant is approximately 50 mg / mL to 100 mg / mL.

[0359] use

[0360] On the other hand, this disclosure provides the use of the above-described empty lipid nanoparticles in the preparation of delivery carriers, which can, for example, deliver active ingredients.

[0361] The hollow lipid nanoparticles disclosed herein possess excellent encapsulation properties for active ingredients. These nanoparticles can serve as carriers to deliver any encapsulated active ingredient to cells, tissues, or organs, offering new therapeutic possibilities for individuals. Understandably, hollow lipid nanoparticles encapsulating active ingredients constitute the aforementioned loaded lipid nanoparticles.

[0362] As used herein, the individual can be any mammal. In some embodiments, the mammal is selected from mice, rats, pigs, cats, dogs, horses, goats, cattle, and monkeys, etc. In some embodiments, the individual is a human.

[0363] On the other hand, this disclosure provides a method for delivering an active ingredient to cells, tissues or organs, including contacting lipid-loaded nanoparticles with cells, tissues or organs.

[0364] Within the scope of this disclosure, the term "cell" is a general term and includes single cells, insect cells, avian cells, fish cells, amphibian cells, mammalian cells, primary cells, continuous cell lines, stem cells, and / or genetically engineered cells.

[0365] In some implementations, the cells are mammalian cells.

[0366] In some implementations, mammalian cells are located within mammalian bodies.

[0367] In some implementation schemes, the tissues or organs are selected from the group consisting of: spleen, liver, kidney, lung, femur, eye tissue, vascular endothelium in blood vessels, lymph nodes, and tumor tissue.

[0368] In another aspect, this disclosure also provides a method for generating peptides and / or proteins of interest in mammalian cells, comprising providing lipid-loaded nanoparticles containing mRNA, contacting cells with the lipid-loaded nanoparticles, wherein after the cells are in contact with the lipid-loaded nanoparticles, the mRNA can be absorbed into the cells and translated to generate peptides and / or proteins of interest.

[0369] Furthermore, this disclosure provides the use of the above-mentioned lipid-loaded nanoparticles or pharmaceutical compositions in the treatment and / or prevention of diseases or conditions.

[0370] In another aspect, this disclosure provides a method for treating and / or preventing diseases or conditions in mammals in need, comprising administering to the mammal a therapeutically and / or preventively effective amount of the aforementioned loaded lipid nanoparticles or pharmaceutical composition.

[0371] As used herein, "effective amount" or "therapeutic and / or preventive effective amount" refers to the amount of the lipid-loaded nanoparticles or pharmaceutical composition of this disclosure that, when administered to a mammal (preferably a human), is sufficient to achieve therapeutic and / or preventive effects in the mammal (preferably a human). The amount of the lipid-loaded nanoparticles or pharmaceutical composition of this disclosure constituting a "therapeutic and / or preventive effective amount" may also be conventionally determined by those skilled in the art based on their own knowledge and the content of this disclosure, for example, depending on the active ingredient and aminolipid compound contained therein, the condition and its severity, the method of administration, and the age of the mammal to be treated.

[0372] Furthermore, this disclosure provides the use of the above-mentioned lipid-loaded nanoparticles or pharmaceutical compositions in the preparation of pharmaceuticals.

[0373] Furthermore, the lipid-loaded nanoparticles or pharmaceutical compositions disclosed herein can be used as a drug.

[0374] In another aspect, this disclosure provides a drug comprising the above-described loaded lipid nanoparticles or pharmaceutical composition.

[0375] In some implementations, the drug is used to treat and / or prevent diseases or conditions.

[0376] In some implementations, the drug is used for gene therapy, protein replacement therapy, antisense therapy, or treatment via interfering RNA, as well as gene vaccination.

[0377] In some implementations, gene vaccination is used to treat and / or prevent cancer, allergies, toxicities, and pathogen infections. Pathogens may be, for example, but are not limited to, one or more selected from viruses, bacteria, and fungi.

[0378] In some implementations, the disease or condition is selected from the group consisting of: rare diseases, infectious diseases, cancer, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases. Among these, cancer may include, but is not limited to, one or more of lung cancer, stomach cancer, liver cancer, esophageal cancer, colon cancer, pancreatic cancer, brain cancer, lymphoma, leukemia, and prostate cancer; genetic diseases may include, but are not limited to, one or more of hemophilia, thalassemia, and Gaucher's disease.

[0379] In some implementations, the active ingredient in the lipid-loaded nanoparticles is a pharmaceutically active ingredient.

[0380] In some implementations, the active pharmaceutical ingredient is a bioactive ingredient, such as a nucleic acid.

[0381] In some embodiments, the nucleic acid is selected from one or more of RNA, antisense oligonucleotides, and DNA. RNA is, for example, messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single-stranded guide RNA (sgRNA), Cas9 mRNA, or mixtures thereof. DNA is, for example, a plasmid.

[0382] In some embodiments, the mass ratio of nucleic acid to aminolipid compound in the above-mentioned drug is about 1:(5-30), for example, about 1:(8-25), 1:(10-20), 1:(13-18), 1:(5-25), 1:(5-20), 1:(5-15), 1:(10-30), 1:(10-25), 1:(10-20), 1:(15-30), 1:(15-25), 1:(20-30), 1:(5-10), 1:(10-15), 1:(15-20), 1:(20-25), or 1:(25-30). For example, the mass ratio of nucleic acid to aminolipid compound in the above-mentioned drug is about 1:10.

[0383] Preparation method:

[0384] Furthermore, the lipid nanoparticles (empty lipid nanoparticles or loaded lipid nanoparticles) or pharmaceutical compositions disclosed herein can be prepared according to methods known in the art. For example, the method may include the following steps:

[0385] (1) Ingredients: Prepare a suitable aqueous phase; and prepare an organic phase comprising the amino lipid compounds of the present disclosure and optionally present auxiliary lipids, structural lipids and / or PEG-lipids;

[0386] (2) Encapsulation: Mix an appropriate amount of aqueous phase with the organic phase;

[0387] (3) Dialysis: Optionally, the mixture from step (2) is dialyzed;

[0388] (4) Sterilization: Optionally, the product of step (3) is sterilized, for example by passing it through a sterilization filter, such as a 0.22 μm microporous membrane.

[0389] In some embodiments, the disclosed lipid nanoparticles or pharmaceutical compositions containing nucleic acids, particularly mRNA, can be prepared by a method comprising the following steps:

[0390] (1) Ingredients: Prepare an aqueous phase containing nucleic acids; and prepare an organic phase (e.g., an ethanol phase) containing amino lipid compounds of the present disclosure and optionally present accessory lipids, structural lipids and / or PEG-lipids;

[0391] (2) Encapsulation: Mix an appropriate amount of aqueous phase with the organic phase;

[0392] (3) Dialysis: Optionally, the mixture from step (2) is dialyzed;

[0393] (4) Sterilization: Optionally, the product of step (3) is sterilized, for example by passing it through a sterilization filter, such as a 0.22 μm microporous membrane. Beneficial effects

[0394] The aminolipid compounds disclosed herein can form delivery carriers with excellent biological activity, such as lipid nanoparticles, which can be used for the delivery of bioactive ingredients, especially water-poorly soluble agents or easily decomposed and degraded active ingredients (e.g., nucleic acids), to improve their bioavailability, or immune activity, or transfection efficiency (for nucleic acids), or safety, or tissue and / or cell targeting or specificity.

[0395] The delivery capability and safety of lipid nanoparticles containing the amino lipid compounds of this disclosure can be determined by methods known in the art.

[0396] The delivery capability of lipid nanoparticles can be determined, for example, by preparing lipid nanoparticles loaded with luciferase (Luc) mRNA and administering them intravenously or intramuscularly to mice, and evaluating the resulting protein expression, for example, by detecting protein expression using an in vivo imaging system (IVIS).

[0397] To make the objectives, technical solutions, and advantages of this invention clearer, the invention is described below with reference to specific embodiments. These embodiments are merely illustrative and not intended to limit the scope of the invention.

[0398] Example

[0399] The following embodiments are provided for illustrative purposes and are not intended to be limiting.

[0400] Experimental methods not specified in the examples are generally under standard conditions or according to the conditions recommended by the raw material or product manufacturer; reagents not specified in the examples are generally commercially available reagents.

[0401] The abbreviations used in the examples have the following meanings: Pd / C Palladium / carbon; EA Ethyl acetate; DCM Dichloromethane; ACN Acetonitrile TEA Triethylamine; H2 Hydrogen MPa Megapascals; DMF N,N-Dimethylformamide; DMSO Dimethyl sulfoxide; EDCI 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; DMAP 4-Dimethylaminopyridine; TBAF Tetrabutylammonium fluoride; TEMPO 2,2,6,6-Tetramethylpiperidine nitride; DCCNa Sodium dichloroisocyanurate; MTBE Methyl tert-butyl ether; THF Tetrahydrofuran; DTN 10-oxononadecanedioic acid; BH3-THF Boranetetrahydrofuran solution; TsOH p-Toluenesulfonic acid; mL Milliliters; h Hours; min Minutes.

[0402] Example 1 Synthesis of aminolipid compound 1-01

[0403] Step 1: Synthesize compound 1-01-A

[0404] Experimental procedure:

[0405] 4-(2-hydroxyethyl)-2,2-dimethyl-1,3-dioxolane (5 g, 34.2 mmol), DCM (50 mL), and TEA (11.4 g, 113.4 mmol) were added to a 250 mL flask. Methanesulfonic anhydride (13.2 g, 75.6 mmol) was slowly added dropwise at 0 °C. After the addition was complete, the mixture was stirred for 20 min and then allowed to react at room temperature for 5 h. A 10% potassium bicarbonate aqueous solution (50 mL) was added, and the mixture was extracted and separated. The organic phase was collected, and the aqueous phase was extracted again with DCM (50 mL). The organic phases were combined and concentrated under reduced pressure to give 8.2 g of crude 1-01-A. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (4:1 v / v) to give 6 g of compound 1-01-A, with a yield of 75.5%.

[0406] Step 2: Synthesis of 4,4-dihexyloxy-1-TBSbutanol

[0407] Experimental procedure:

[0408] 4-(tert-butyldimethylsilyl)oxo-1-butanal (10.0 g, 49.4 mmol), MTBE (40 mL), cyclohexane (40 mL), n-hexanol (12.6 g, 123.5 mmol), and ammonium bromide (242 mg, 2.47 mmol) were added to a 250 mL flask, and the mixture was stirred and refluxed for 6 h to separate the water. The mixture was filtered, and the filtrate was collected and concentrated under reduced pressure to obtain crude 4,4-dihexyloxy-1-TBSbutanol. The crude product was purified by silica gel column chromatography, eluting with n-heptane, to give 16.2 g of 4,4-dihexyloxy-1-TBSbutanol, in 84.5% yield.

[0409] Step 3: Synthesize compound BSB

[0410] Experimental procedure:

[0411] 4,4-Dihexyloxy-1-TBSbutanol (16.2 g, 41.7 mmol), THF (80 mL), and TBAF (13.1 g, 41.7 mmol) were added to a 250 mL flask, and the mixture was stirred and reacted at room temperature for 4 h. A saturated sodium bicarbonate solution (80 mL) was added, and the mixture was extracted twice with EA (80 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude 4,4-dihexyloxy-1-butanol. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (5:1 v / v) to give compound BSB 10.1 g, in 88.6% yield.

[0412] Step 4: Synthesize compound 1-01-B

[0413] Experimental procedure:

[0414] (1) Compound DNT was prepared according to CN114805049A.

[0415] (2) EDCI (8.7 g, 45.5 mmol), DCM (60 mL), pyridine (4.3 g, 59.6 mmol), DMAP (0.67 g, 5.5 mmol), BSB (10 g, 36.4 mmol), and DTN (6.2 g, 18.2 mmol) were added sequentially to a 100 mL flask, and the reaction was carried out at room temperature for 4 h. Water (100 mL) was added, and the mixture was stirred and extracted. The organic phase was collected, and the aqueous phase was extracted with DCM (50 mL). The organic phases were combined. The mixture was concentrated under reduced pressure to obtain 17.8 g of crude compound 1-01-B. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v, 20:1) to obtain 16 g of compound 1-01-B, with a yield of 82.2%.

[0416] Step 5: Synthesize compound 1-01-C

[0417] Experimental procedure:

[0418] Compound 1-01-B (3.44 g, 4.0 mmol), MTBE (15 mL), cyclohexane (15 mL), compound 1-01-A (1.8 g, 8.6 mmol), and TsOH (15.2 mg, 0.088 mmol) were added sequentially to a 100 mL flask, and the mixture was reacted at 100 °C for 4 h. After cooling to room temperature, 5% potassium bicarbonate aqueous solution (15 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted with EA (15 mL), and the organic phases were combined and concentrated under reduced pressure to obtain 4.6 g of crude compound 1-01-C. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v ratio 20:1) to obtain 500 mg of compound 1-01-C, with a yield of 12.2%.

[0419] Step 6: Synthesize compound 1-01

[0420] Experimental procedure:

[0421] Compound 1-01-C (0.5 g, 0.49 mmol) and dimethylaminetetrahydrofuran solution (25 mL, 1.0 mol / L) were added sequentially to a 25 mL flask, and the reaction was carried out at room temperature for 48 h. The reaction solution was concentrated under reduced pressure to obtain 600 mg of crude compound 1-01. The crude product was purified by reverse-phase extraction using ethanol and water (ethanol volume percentage 65%–100%) to give 250 mg of pale yellow oily compound 1-01 product, with a yield of 52.6% and a purity of 95.23%.

[0422] 1H NMR (600MHz, CDCl3) δ4.48(t,J=5.2Hz,2H),4.12-4.02(m,6H),3.56(dt,J=9.2,6.7Hz,4H),3.48(m,J=7.5Hz,1H),3.40(dt,J=9.3,6.7Hz,4H),2 .41(dd,J=39.5,31.8Hz,2H),2.28(dd,J=14.9,7.4Hz,10H),1.71-1.65 (m,8H),1.61-1.52(m,16H),1.36-1.23(m,46H),0.89(t,J=7.0Hz,12H).

[0423] LC-MS(ESI):Calculated for 970.51,Found(M+H):971.2

[0424] Example 2 Synthesis of aminolipid compound 1-06

[0425] Compound 1-06-C was synthesized according to steps one through five of Example 1, and then compound 1-06 was synthesized according to step six of Example 1, wherein compound 1-06-C (0.7 g, 0.71 mmol) and dimethylaminetetrahydrofuran solution (25 mL, 2.0 mol / L) yielded 0.36 g of pale yellow oily compound 1-06 product, with a yield of 54.6% and a purity of 93.37%.

[0426] 1 H NMR (600MHz, CDCl3) δ4.48 (t, J=5.2Hz, 2H), 4.11-4.04 (m, 6H), 3.56 (dt, J=9 .2,6.7Hz,4H),3.47(m,J=5.4Hz,1H),3.40(dt,J=9.2,6.7Hz,4H),2.39(dd, J=12.0,9.7,5.5Hz,2H),2.30-2.26(m,4H),2.22(s,6H),1.70-1.65(m,8H), 1.58-1.54(m,22H),1.31(m,J=9.2,5.6,5.0Hz,34H),0.89(t,J=6.9Hz,12H).

[0427] LC-MS(ESI):Calculated for 928.43,Found(M+H):929.1

[0428] Example 3 Synthesis of aminolipid compound 1-07

[0429] Compound 1-07-C was synthesized according to steps one through five of Example 1, and then compound 1-07 was synthesized according to step six of Example 1, wherein compound 1-07-C (0.65 g, 0.61 mmol) and dimethylaminetetrahydrofuran solution (25 mL, 2.0 mol / L) yielded 0.26 g of pale yellow oily compound 1-07 product, with a yield of 45.61% and a purity of 94.89%.

[0430] 1 H NMR (600MHz, CDCl3) δ4.46(t,J=5.2Hz,2H),4.09-4.01(m,6H),3.55(dt,J=9.2,6.7Hz,4H),3.46(t,J=7.6Hz,1H),3.39(dt,J=9.3 ,6.7Hz,4H),2.26(t,J=7.6Hz,4H),2.22(s,6H),1.70-1.62(m,10H),1.61-1.51(m,16H),1.37-1.21(m,42H),0.90-0.84(m,12H).

[0431] LC-MS(ESI):Calculated for 942.46,Found(M+H):943.3

[0432] Example 4 Synthesis of aminolipid compound 1-09

[0433] Compound 1-09-C was synthesized according to steps one through five of Example 1, and then compound 1-09 was synthesized according to step six of Example 1, wherein compound 1-09-C (0.95 g, 1.01 mmol) and dimethylaminetetrahydrofuran solution (25 mL, 2.0 mol / L) yielded 0.29 g of pale yellow oily compound 1-09 product, with a yield of 32.4% and a purity of 97.37%.

[0434] 1 H NMR (600MHz, CDCl3) δ4.52(t,J=5.3Hz,2H),4.16-4.07(m,6H),3.61(dt,J=9.3,6.7Hz,4H),3.52(m,J=7.5Hz,1H),3.45(dt,J=9.3,6.7H z,4H),2.50(d,J=41.4Hz,2H),2.42-2.23(m,10H),1.86-1.69(m,10H),1.68-1.56(m,16H),1.45-1.28(m,32H),0.93(t,J=7.0Hz,12H).

[0435] LC-MS(ESI):Calculated for 886.35,Found(M+H):887.0

[0436] Example 5 Synthesis of aminolipid compounds 1-32

[0437] Step 1: Synthesis of compound 1-32-G

[0438] Experimental procedure:

[0439] 4-(2-hydroxyethyl)-2,2-dimethyl-1,3-dioxolane (10 g, 68.40 mmol), DCM (60 mL), and TEA (13.84 g, 136.81 mmol) were added sequentially to a 250 mL dry flask. The mixture was stirred at 0 °C, and p-toluenesulfonic anhydride (29 g, 88.94 mmol) was added. After the addition was complete, the mixture was allowed to react at room temperature for 3 h. Water (50 mL) was added and stirred. The mixture was separated, and the aqueous phase was extracted with DCM (50 mL). The organic phases were combined and evaporated to dryness under reduced pressure to obtain 24 g of crude compound 1-32-G. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v ratio 4:1) to obtain 14 g of compound 1-32-G, with a yield of 68.14%.

[0440] Step 2: Synthesizing BSA

[0441] Experimental procedure:

[0442] Add BSB (4.00 g, 14.57 mmol, as synthesized in step 3 of Example 1), DCM (32 mL), 2,2,6,6-tetramethylpiperidine oxide (114 mg, 0.73 mmol), sodium bromide (150 mg, 1.46 mmol), and potassium bicarbonate (1.46 g, 14.57 mmol) to a 100 mL flask. Maintain the temperature at -5 ± 5 °C and add DCCNa solution (2.24 g dissolved in 24 mL of water, 10.20 mmol) dropwise. After the addition is complete, allow the reaction to proceed for 19 h. Filter, add water (24 mL) to the filtrate, and extract and separate the liquid. Wash the organic phase with water (24 mL), concentrate the organic phase under reduced pressure, and give compound BSA 4.58 g, yield 109%.

[0443] Step 3: Synthesize compound 1-32-B

[0444] Experimental procedure:

[0445] (1) Compound 1-32-A was prepared according to CN114805049A.

[0446] (2) To a 250 mL dry flask, EDCI (18.55 g, 38.71 mmol), pyridine (9.17 g, 116.14 mmol), DCM (60 mL), DMAP (1.42 g, 11.61 mmol), benzyl alcohol (7.95 g, 73.56 mmol), and compound 1-32-A (10 g, 38.71 mmol) were added sequentially, and the mixture was reacted overnight at room temperature for 16 h. DCM (50 mL) and water (50 mL) were added, and the mixture was extracted and separated. The aqueous phase was then extracted again with DCM (50 mL). The organic phases were combined, concentrated under reduced pressure, and purified by silica gel column chromatography. The elution was performed with n-heptane and EA (6:1 v / v) to give 13.12 g of compound 1-32-B, with a yield of 77.27%.

[0447] Step 4: Synthesis of compound 1-32-C

[0448] Experimental procedure:

[0449] To a 250 mL dry flask, compound 1-32-B (6 g, 13.68 mmol), compound 1-32-G (8.22 g, 27.36 mmol), toluene (50 mL), and TsOH (118 mg, 0.68 mmol) were added sequentially, and the mixture was heated to reflux and the water was removed for 8 h. After cooling to room temperature, 30 mL of 5% sodium bicarbonate aqueous solution was added, and the mixture was extracted and separated. The aqueous phase was extracted with EA (40 mL), and the organic phases were combined and evaporated to dryness under reduced pressure to obtain crude compound 1-32-C. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (3:1 v / v) to give 5.8 g of compound 1-32-C, with a yield of 63.53%.

[0450] Step 5: Synthesize compound 1-32-D

[0451] Experimental procedure:

[0452] To a 100 mL dry flask, add compound 1-32-C (6 g, 8.82 mmol) and THF (30 mL), stir at 0 °C, and add 1.0 M boranetetrahydrofuran solution (44 mL) dropwise. After the addition is complete, transfer to room temperature and react for 16 h. Stir the reaction mixture at 0 °C, quench with water dropwise until no bubbles are generated, concentrate the reaction mixture under reduced pressure, add water (100 mL) and DCM (100 mL) to the crude product, extract and separate the phases. Extract the aqueous phase with DCM (100 mL), combine the organic phases, concentrate under reduced pressure to obtain crude compound 1-32-D. Purify the crude product by silica gel column chromatography, eluting with n-heptane and EA (2:1 v / v) to obtain 2 g of compound 1-32-D, yield 48.01%.

[0453] Step 6: Synthesize compound 1-32-E

[0454] Experimental procedure:

[0455] To a 100 mL dry flask, EDCI (1.01 g, 5.29 mmol), pyridine (668.8 mg, 8.46 mmol), DCM (10 mL), DMAP (78 mg, 0.64 mmol), compound 1-32-D (1 g, 2.12 mmol), and BSA (1.53 g, 5.29 mmol) were added sequentially, and the mixture was reacted at room temperature for 4 h. DCM (30 mL) and water (30 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (50 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-32-E. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v, 10:1) to give 1 g of compound 1-32-E, in 46.63% yield.

[0456] Step 7: Synthesize compound 1-32

[0457] Experimental procedure:

[0458] Compound 1-32-E (1 g, 0.99 mmol) and 2.0 M dimethylaminetetrahydrofuran solution (30 mL) were added to a 100 mL dry flask and the mixture was stirred at room temperature for 20 h. The reaction solution was evaporated to dryness under reduced pressure, and water (30 mL) and EA (30 mL) were added. The mixture was extracted and separated, and the aqueous phase was extracted with EA. The organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-32. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v ratio 2:1) to give 280 mg of pale yellow oily compound 1-32 product, with a yield of 32.18% and a purity of 96.96%.

[0459] 1 H NMR (600MHz, CDCl3) δ4.48 (t, J=5.6Hz, 2H), 4.06 (dt, J=23.9, 6.8Hz, 6H), 3.56 (dt, J=9.2,6.7Hz,4H),3.46(t,J=7.6Hz,1H),3.39(dt,J=9.3,6.7Hz,4H),2.37(t,J=7.6 Hz,4H),2.32-2.25(m,1H),2.22(s,6H),1.91(dd,J=13.3,7.5Hz,4H),1.80(m,J=12 .7,9.4,6.2Hz,1H),1.62-1.51(m,16H),1.38-1.21(m,38H),0.88(t,J=6.9Hz,12H).

[0460] LC-MS: Calculated for 886.35, Found(M+H): 886.9.

[0461] Example 6 Synthesis of aminolipid compounds 1-33

[0462] Step 1: Synthesis of compound 1-33-B

[0463] Experimental procedure:

[0464] (1) Compound 1-33-A was prepared according to CN114805049A.

[0465] (2) EDCI (6.68 g, 34.84 mmol), DCM (60 mL), DMAP (4.25 g, 34.84 mmol), undecyl alcohol (5 g, 29.04 mmol), and compound 1-33-A (7-oxotridecanoic acid) (15 g, 58.07 mmol) were added sequentially to a 250 mL dry flask, and the reaction was carried out at room temperature for 15 h. Water (50 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (50 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-33-B. The crude product was purified by silica gel column chromatography, eluting with DCM and methanol (volume ratio 15:1) to obtain 7 g of compound 1-33-B, with a yield of 58.43%.

[0466] Step 2: Synthesize compound 1-33-C

[0467] Experimental procedure:

[0468] Compound 1-33-B (5 g, 12.12 mmol), compound 1-32-G (7.64 g, 25.45 mmol, synthesized according to the first step of Example 5), toluene (50 mL), and TsOH (65 mg, 0.38 mmol) were added to a 100 mL dry flask, and the mixture was refluxed to remove water for 8 h. After cooling to room temperature, 5% sodium bicarbonate solution (30 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted with EA (50 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-33-C. The crude compound was purified by silica gel column chromatography, eluting with DCM and methanol (16:1 v / v) to give 3.5 g of compound 1-33-C, with a yield of 44.08%.

[0469] Step 3: Synthesize compound 1-33-D

[0470] Experimental procedure:

[0471] To a 250 mL dry flask, EDCI (1.22 g, 6.35 mmol), pyridine (580 mg, 7.33 mmol), DMAP (178 mg, 1.47 mmol), DCM (30 mL), BSB (1.34 g, 4.89 mmol, see step 3 of Example 1), and compound 1-33-C (3.2 g, 4.89 mmol) were added sequentially, and the reaction was carried out at room temperature for 16 h. DCM (30 mL) and water (40 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (40 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-33-D. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (8:1 v / v) to give 2 g of compound 1-33-D, with a yield of 44.92%.

[0472] Step 4: Synthesize compound 1-33

[0473] Experimental procedure:

[0474] Compound 1-33-D (2 g) and 2M dimethylaminetetrahydrofuran solution (30 mL) were added to a 100 mL dry flask, and the mixture was stirred at room temperature for 24 h. The mixture was concentrated under reduced pressure to obtain crude compound 1-33. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (2:1 v / v) to give 1.1 g of pale yellow oily compound 1-33, with a yield of 63.95% and a purity of 94.03%.

[0475] 1 H NMR (600MHz, CDCl3) δ4.47(t,J=5.1Hz,1H),4.07(m,J=13.6,11.2,6.9Hz,6H),3.55(dt,J=9.1,6.7Hz,2H),3.49-3.43(m,1H),3.39(dt,J= 9.2, 6.7Hz, 2H), 2.42-2.35 (m, 1H), 2.27 (dd, J=14.5, 7.0Hz, 5H), 2.22 (s, 6H), 1.69-1.53 ​​(m, 18H), 1.38-1.21 (m, 38H), 0.90-0.85 (m, 9H).

[0476] LC-MS: Calculated for 784.22, Found(M+H): 784.8.

[0477] Example 7 Synthesis of aminolipid compounds 1-39

[0478] Step 1: Synthesize compound 1-39-A

[0479] Experimental procedure:

[0480] EDCI (43.5 g, 0.227 mol), DCM (300 mL), DMAP (27.8 g, 0.227 mol), benzyl alcohol (20.5 g, 0.189 mol), and glutaric acid (50 g, 0.378 mol) were added sequentially to a 1000 mL dry flask, and the reaction was carried out at room temperature for 18 h. Water (150 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (200 mL), and the organic phases were combined and concentrated under reduced pressure to give 40 g of compound 1-39-A, with a yield of 95.12%.

[0481] Step 2: Synthesize compound 1-39-B

[0482] Experimental procedure:

[0483] Compound 1-39-A (40 g, 0.18 mol) and THF (100 mL) were added to a 1 L flask. The mixture was cooled to 0 °C, and 1 M boranetetrahydrofuran solution (180 mL, 0.18 mol) was slowly added dropwise. After the addition was complete, the mixture was allowed to react at room temperature for 1.5 h. The mixture was then cooled to 0 °C, and the reaction was quenched by adding water (40 mL). The mixture was concentrated under reduced pressure to obtain crude compound 1-39-B. The crude compound was extracted with water (200 mL) and EA (100 mL), and the extraction was repeated three times. The organic phases were combined, concentrated under reduced pressure, and purified by silica gel column chromatography. The mixture was eluted with n-heptane and EA (2:1 v / v) to give 25 g of compound 1-39-B, with a yield of 66.7%.

[0484] Step 3: Synthesize compound 1-39-C

[0485] Experimental procedure:

[0486] To a 500 mL dry flask, compound 1-39-B (25 g, 120.04 mmol), DCM (100 mL), TEMPO (566 mg, 3.63 mmol), sodium bromide (377 mg, 3.63 mmol), and potassium bicarbonate (9.62 g, 96.04 mmol) were added sequentially. The mixture was cooled to 0 °C and stirred. DCCNa (17.16 g, 78.03 mmol, dissolved in 120 mL water) solution was slowly added dropwise. After the addition was complete, the reaction mixture was allowed to react at low temperature for 2 h. The reaction solution was filtered, and the filtrate was separated. The aqueous phase was extracted with DCM (100 mL). The organic phases were combined and distilled under reduced pressure to obtain crude compound 1-39-C. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (6:1 v / v) to give 18 g of compound 1-39-C, with a yield of 72.69%.

[0487] Step 4: Synthesize compound 1-39-D

[0488] Experimental procedure:

[0489] To a 500 mL dry flask, compound 1-39-C (17 g, 82.43 mmol), ammonium bromide (1.61 g, 16.49 mmol), n-hexanol (16.84 g, 164.86 mmol), cyclohexane (50 mL), and MTBE (100 mL) were added sequentially, and the mixture was refluxed for 8 h to separate the water. A 5% potassium bicarbonate aqueous solution (40 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted with EA, and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-39-D. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (20:1 v / v) to give 25 g of compound 1-39-D, with a yield of 77.26%.

[0490] Step 5: Synthesize compound 1-39-E

[0491] Experimental procedure:

[0492] To a 50 mL dry flask, compound 1-39-D (2.5 g, 6.37 mmol), THF (15 mL), and Pd / C (500 mg) were added sequentially. The mixture was purged with hydrogen three times and reacted at room temperature for 18 h. The reaction solution was filtered and washed with THF (20 mL) to obtain a solution of compound 1-39-E.

[0493] Step 6: Synthesize compound 1-39-F

[0494] Experimental procedure:

[0495] To a 100 mL dry flask, EDCI (1.01 g, 5.29 mmol), pyridine (668.8 mg, 8.46 mmol), DCM (10 mL), DMAP (78 mg, 0.64 mmol), compound 1-32-D (1 g, 2.12 mmol), and a solution of compound 1-39-E obtained in step 5 were added sequentially, and the mixture was reacted at room temperature for 4 h. DCM (30 mL) and water (30 mL) were added to the reaction mixture, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (30 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 1-39-F. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v, 12:1) to give 1 g of compound 1-39-F, in 45.34% yield.

[0496] Step 7: Synthesize compound 1-39

[0497] Experimental procedure:

[0498] Compound 1-39-F (800 mg, 0.77 mmol) and 2.0 M dimethylaminetetrahydrofuran solution (30 mL) were added to a 100 mL dry flask, and the mixture was stirred at room temperature for 18 h. The reaction solution was distilled under reduced pressure to obtain crude compound 1-39. EA (30 mL) and water (30 mL) were added to the crude product, and the mixture was extracted and separated. The aqueous phase was extracted with EA (30 mL), and the organic phases were combined and concentrated under reduced pressure. After reverse-phase purification, elution with ethanol and water (ethanol volume percentage 85%–100%) yielded 160 mg of a pale yellow oily product of compound 1-39, with a purity of 94.27% and a yield of 22.86%.

[0499] 1 H NMR (600MHz, CDCl3) δ4.46(t,J=5.4Hz,2H),4.12-3.99(m,6H),3.56(dt,J=9.2,6.7Hz,4H),3.48(t,J=7.5Hz,1H),3.39(d t,J=9.2,6.7Hz,4H),2.32(t,J=7.2Hz,4H),2.27(s,6H),1.71-1.52(m,24H),1.40-1.21(m,40H),0.88(t,J=6.9Hz,12H).

[0500] LC-MS: Calculated for 914.40, Found(M+H): 915.0.

[0501] Example 8 Synthesis of aminolipid compound 2A-01

[0502] Step 1: Synthesis of compound 3301-E

[0503] Experimental procedure:

[0504] 3-Amino-1-propanol (10 g, 133.14 mmol), TEA (11.2 g, 110.95 mmol), phthalic anhydride (16.4 g, 110.95 mmol), and toluene (80 mL) were added to a 250 mL dry flask, and the mixture was refluxed for 5 h. The reaction solution was concentrated under reduced pressure to obtain crude compound 3301-E. DCM (100 mL) and water (50 mL) were added to the crude product, and the mixture was extracted and separated. The organic phase was washed again with water (50 mL), and the organic phase was concentrated under reduced pressure to obtain 18 g of compound 3301-E, with a yield of 79.05%.

[0505] Step 2: Synthesize compound 3301-F

[0506] Experimental procedure:

[0507] Compound 3301-E (18 g, 87.71 mmol), DCM (100 mL), TEMPO (414 mg, 2.65 mmol), and sodium bromide (272 mg, 2.65 mmol) were added to a 250 mL dry flask. The mixture was stirred at 0 °C, and DCCNa solution (DCCNa (12.54 g, 57.01 mmol) dissolved in 100 mL of water) was slowly added dropwise. After the addition was complete, the mixture was stirred and kept at this temperature for 2 h. The reaction mixture was filtered, and the filtrate was separated. The aqueous phase was extracted with DCM (50 mL), and the DCM phases were combined and concentrated under reduced pressure to obtain crude compound 3301-F. The crude product was purified by silica gel column chromatography, eluting with DCM, to obtain 10.4 g of compound 3301-F, with a yield of 58.36%.

[0508] Step 3: Synthesize compound 2A-01-A

[0509] Experimental procedure:

[0510] Add EDCI (53.18 g, 0.277 mol), DMAP (33.89 g, 0.277 mol), and DCM (350 mL) to a 1000 mL dry flask, stir for 5 min, then add azelaic acid (87.03 g, 0.462 mol) and benzyl alcohol (25 g, 0.231 mol), and react at room temperature for 2 h. Add water (300 mL), extract and separate the liquid phase. Extract the aqueous phase with DCM (200 mL), combine the organic phases, add water (200 mL), extract and separate the liquid phase, concentrate the organic phase under reduced pressure to give 65.5 g of compound 2A-01-A, yield 90.88%.

[0511] Step 4: Synthesize compound 2A-01-B

[0512] Experimental procedure:

[0513] Compound 2A-01-A (65.5 g, 235.32 mmol) and THF (130 mL) were added to a 1000 mL dry flask. The mixture was stirred at 0 °C, and 1.0 M boranetetrahydrofuran solution (240 mL, 240 mmol) was added dropwise. After the addition was complete, the mixture was allowed to react at room temperature for 2 h. The reaction was quenched by adding saturated saline (100 mL). The mixture was separated, and the aqueous phase was extracted with EA (200 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01-B. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (8:1 v / v) to give 43.6 g of compound 2A-01-B, with a yield of 70.08%.

[0514] Step 5: Synthesize compound 2A-01-C

[0515] Experimental procedure:

[0516] Compound 2A-01-B (43.6 g, 164.92 mmol), DCM (240 mL), potassium bicarbonate (16.51 g, 164.92 mmol), sodium bromide (0.85 g, 8.25 mmol), and TEMPO (1.29 g, 8.25 mmol) were added to a 1 L dry flask. The mixture was stirred at 5 °C, and an aqueous solution of DCCNa (21.76 g, 98.95 mmol) (200 mL) was added dropwise. The reaction was allowed to proceed for 1.5 h after the addition was complete. The reaction mixture was filtered, and the aqueous phase was extracted twice with DCM (200 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01-C. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v, 10:1) to give 29.66 g of compound 2A-01-C, with a yield of 68.55%.

[0517] Step 6: Synthesize compound 2A-01-D

[0518] Experimental procedure:

[0519] EDCI (17.92 g, 93.55 mmol), pyridine (9.24 g, 116.94 mmol), DCM (160 mL), DMAP (2.85 g, 23.39 mmol), benzyl alcohol (7.59 g, 70.16 mmol), and 8-bromooctanoic acid (17.39 g, 77.96 mmol) were added to a 500 mL flask, and the reaction was carried out at room temperature for 16 h. The reaction was quenched with water (50 mL), and the mixture was separated. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined and concentrated under reduced pressure to give crude compound 2A-01-D. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v, 15:1) to give 16.5 g of compound 2A-01-D, with a yield of 68.58%.

[0520] Step 7: Synthesize compound 2A-01-E

[0521] Experimental procedure:

[0522] Compound 2A-01-D (24.3 g, 77.58 mmol), toluene (200 mL), and triphenylphosphine (22.38 g, 85.34 mmol) were added to a 1 L dry flask, and the mixture was refluxed for 50 h. After cooling, the mixture was concentrated under reduced pressure to obtain crude compound 2A-01-E. The crude product was purified by silica gel column chromatography, eluting with DCM and methanol (30:1 v / v) to give 29.12 g of compound 2A-01-E, with a yield of 65.28%.

[0523] Step 8: Synthesize compound 2A-01-F

[0524] Experimental procedure:

[0525] Compound 2A-01-E (19 g, 33.01 mmol), DMSO (60 mL), 2A-01-C (6.1 g, 23.55 mmol), and potassium carbonate (4.9 g, 35.37 mmol) were added sequentially to a 250 mL dry flask, and the mixture was heated to 100 °C and reacted for 6 h. After cooling to room temperature, water (150 mL) was added, and the mixture was extracted and separated. The aqueous phase was extracted twice with EA (50 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01-F. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (v / v ratio 12:1) to give 3 g of compound 2A-01-F, with a yield of 26.58%.

[0526] Step 9: Synthesize compound 2A-01-G

[0527] Experimental procedure:

[0528] Compound 2A-01-F (2.8 g, 5.85 mmol), DCM (14 mL), methylrhenium trioxide (15 mg, 0.06 mmol), and 30% hydrogen peroxide solution (663 mg, 5.85 mmol) were added to a 100 mL flask. The mixture was purged with nitrogen three times and reacted at room temperature. Then, DCM (20 mL) and water (20 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (30 mL), and the organic phases were combined. The solvent was evaporated to dryness under reduced pressure. Acetonitrile (18 mL) and water (6 mL) were then added, followed by slow dropwise addition of dilute sulfuric acid (3.6 M, 1.5 mL). After the addition was complete, the mixture was stirred at room temperature for 4 h. Add 20 mL of 5% potassium bicarbonate aqueous solution and 20 mL of EA, extract and separate the phases, extract the aqueous phase with EA (30 mL), combine the organic phases, concentrate under reduced pressure to obtain crude compound 2A-01-G. The crude product is purified by silica gel column chromatography, eluted with n-heptane and EA (volume ratio 3:1), to obtain 2.5 g of compound 2A-01-G, yield 83.33%.

[0529] Step 10: Synthesize compound 2A-01-H

[0530] Experimental procedure:

[0531] Compound 2A-01-G (3 g, 5.85 mmol), compound 3301-F (1.56 g, 7.62 mmol), toluene (40 mL), and TsOH (51.6 mg, 0.3 mmol) were added sequentially to a 100 mL flask, and the mixture was refluxed to remove water for 4 h. The mixture was cooled to room temperature, and 30 mL of 5% potassium bicarbonate aqueous solution was added. The mixture was extracted and separated. The aqueous phase was extracted with EA (30 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01-H. The crude compound was purified by silica gel column chromatography, eluting with n-heptane and EA (7:1 v / v) to give 3.5 g of compound 2A-01-H, with a yield of 85.78%.

[0532] Step 11: Synthesize compound 2A-01-I

[0533] Experimental procedure:

[0534] Compound 2A-01-H (3.5 g, 5.02 mmol), THF (20 mL), and Pd / C (700 mg) were added to a 100 mL dry flask. The mixture was purged with hydrogen three times and reacted at room temperature for 20 h. The reaction solution was filtered, and the filter cake was washed twice with THF (20 mL) to obtain a tetrahydrofuran solution of compound 2A-01-I.

[0535] Step 12: Synthesize compound 2A-01-J

[0536] Experimental procedure:

[0537] Add EDCI (2.31 g, 12.08 mmol), pyridine (1.14 g, 14.49 mmol), DCM (20 mL), DMAP (177 mg, 1.45 mmol), BSB (2.51 g, 9.18 mmol, see step 3 of Example 1), and a tetrahydrofuran solution of compound 2A-01-I obtained in step 11 to a 100 mL dry flask, and react at room temperature for 16 h. Add DCM (30 mL) and water (30 mL), extract and separate the phases. Extract the aqueous phase with DCM (30 mL), combine the organic phases, concentrate under reduced pressure to obtain crude compound 2A-01-J. Purify the crude product by silica gel column chromatography, eluting with n-heptane and EA (v / v ratio 10:1) to obtain 2.8 g of compound 2A-01-J, yield 56.22%.

[0538] Step 13: Synthesize compound 2A-01-K

[0539] Experimental procedure:

[0540] Compound 2A-01-J (2.8 g, 2.72 mmol), anhydrous ethanol (30 mL), and hydrazine hydrate (1.36 g, 13.59 mmol) were added to a 100 mL dry flask, and the mixture was refluxed for 2 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. DCM (60 mL) and water (30 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (30 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01-K. The crude product was purified by silica gel column chromatography, eluting with DCM and methanol (v / v, 10:1) to give 2.2 g of compound 2A-01-K, with a yield of 89.79%.

[0541] Step 14: Synthesize compound 2A-01

[0542] Experimental procedure:

[0543] Compound 2A-01-K (2.2 g, 2.44 mmol), anhydrous ethanol (15 mL), Pd / C (440 mg), and 37% formaldehyde aqueous solution (1 g, 12.22 mmol) were added to a 100 mL dry flask. The mixture was purged with hydrogen three times and reacted at room temperature for 16 h. The reaction solution was filtered, and the filtrate was evaporated to dryness. DCM (50 mL) and water (30 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (30 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-01. The crude product was purified by reverse-phase extraction using ethanol and water (ethanol volume percentage 85%–100%) to give 670 mg of pale yellow oily compound 2A-01 product with a purity of 95.96% and a yield of 29.52%.

[0544] 1 H NMR (600MHz, CDCl3) δ5.00(t,J=4.8Hz,1H),4.47(t,J=5.2Hz,2H),4.07(t,J=6.1H z,4H),3.56(dt,J=9.2,6.7Hz,6H),3.40(dt,J=9.2,6.7Hz,4H),2.41(dd,J=18.0, 9.8Hz,2H),2.30-2.25(m,4H),2.24(s,6H),1.85-1.79(m,2H),1.72-1.63(m,8H), 1.64-1.58(m,4H),1.58-1.48(m,12H),1.37-1.23(m,38H),0.88(t,J=6.9Hz,12H).

[0545] LC-MS: Calculated for 928.43, Found(M+H): 929.2.

[0546] Example 9 Synthesis of aminolipid compound 2A-09

[0547] Experimental procedure:

[0548] Compound 2A-09-K was synthesized according to steps one through thirteen of Example 8, and then compound 2A-09 was synthesized according to step fourteen of Example 8, wherein 2A-09-K (0.6 g, 0.66 mmol), 37% formaldehyde aqueous solution (266 mg, 3.28 mmol), Pd / C (120 mg), and anhydrous ethanol (6 mL) were purified to obtain 200 mg of pale yellow oily compound 2A-09 product with a purity of 95.79% and a yield of 32.26%.

[0549] 1 H NMR (600MHz, CDCl3) δ5.01(t,J=4.8Hz,1H),4.47(t,J=5.2Hz,2H),4.07(t,J =6.1Hz,4H),3.56(dt,J=9.2,6.7Hz,6H),3.40(dt,J=9.2,6.7Hz,4H),2.40(d ,J=23.5Hz,2H),2.28(t,J=7.6Hz,4H),2.25(s,6H),1.86-1.80(m,2H),1.72- 1.65(m,8H),1.64-1.50(m,14H),1.37-1.25(m,42H),0.88(t,J=6.9Hz,12H).

[0550] LC-MS: Calculated for 942.46, Found(M+H): 943.0.

[0551] Example 10 Synthesis of aminolipid compound 2A-29

[0552] Step 1: Synthesize compound 2A-29-A

[0553] Experimental process

[0554] EDCI (35.91 g, 187.30 mmol), pyridine (19.75 g, 249.74 mmol), and DCM (200 mL) were added to a 500 mL flask and stirred at room temperature for 10 min. Then, 1,7-pimercic acid (20.00 g, 124.87 mmol), DMAP (3.05 g, 24.97 mmol), and benzyl alcohol (13.50 g, 124.87 mmol) were added, and the mixture was stirred at room temperature for 3 h. Water (200 mL) was added, and the mixture was stirred for 5 min before separation. The organic phase was collected. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined and concentrated under reduced pressure to give compound 2A-29-A29.00 g.

[0555] Step 2: Synthesize compound 2A-29-B

[0556] Experimental procedure:

[0557] Compound 2A-29-A (29.00 g, 115.86 mmol) and THF (116 mL) were added to a 1 L flask. The mixture was stirred and cooled to 0 ± 5 °C. A 1 M boranetetrahydrofuran solution (116 mL, 116 mmol) was added dropwise under controlled temperature. After the addition was complete, the mixture was heated to 25 ± 5 °C and reacted for 2 h. The reaction was quenched by adding water (116 mL) dropwise below 0 °C. Saturated saline solution (116 mL) was then added, and the mixture was stirred for 5 min. The aqueous phase was separated. The aqueous phase was extracted twice with EA (116 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-29-B. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v ratio 1:5) to give 11.00 g of compound 2A-29-B, with a yield of 48.7%.

[0558] Step 3: Synthesize compound 2A-29-C

[0559] Experimental procedure:

[0560] Compound 2A-29-B (11.0 g, 46.55 mmol) and DCM (110 mL) were added to a 250 mL flask and stirred until dissolved. The temperature was kept below 30 °C, and Desmond reagent (23.69 g, 55.86 mmol) was added in portions to the reaction mixture. The reaction was carried out at 25 ± 5 °C for 3 h. Water (110 mL) was added and stirred for 5 min. Insoluble matter was removed by filtration. The filtrate was separated, and the organic phase was collected and concentrated under reduced pressure to obtain crude compound 2A-29-C. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v, 1:9) to give 4.60 g of compound 2A-29-C, in 42.2% yield.

[0561] Step 4: Synthesize compound 2A-29-D

[0562] Experimental procedure:

[0563] Compound 2A-29-C (3.90 g, 16.65 mmol), benzyl alcohol (3.9 mL), 3-benzyl-5-(2-hydroxyethyl)-4-methyl-1,3-thiazolyl chloride (539 mg, 2.00 mmol), and TEA (5.31 g, 52.44 mmol) were added to a 100 mL flask. The reaction was carried out at 80 ± 5 °C for 2 h. DCM (39 mL) and water (39 mL) were added, and the mixture was extracted and separated. The organic phase was washed twice with water (39 mL), collected, and concentrated under reduced pressure to give crude compound 2A-29-D. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v, 1:4) to give 3.00 g of compound 2A-29-D, yield 76.9%.

[0564] Step 5: Synthesize compound 2A-29-E

[0565] Experimental procedure:

[0566] Compound 2A-29-D (3.0 g, 6.40 mmol) and DCM (10.5 mL) were added to a 50 mL flask. NaBH4 (194 mg, 5.12 mmol) was added in portions at -5 ± 5 °C. After the addition was complete, methanol (1.5 mL) was added dropwise to the reaction mixture. The reaction was maintained at -5 ± 5 °C for 2 h. A saturated NH4Cl solution (10.5 mL) was added dropwise, and the mixture was stirred for 10 min. The organic phase was collected by separation. The aqueous phase was extracted once with DCM (10.5 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-29-E. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (3:7 v / v) to give 3.00 g of compound 2A-29-E, with a yield of 99.7%.

[0567] Step 6: Synthesize compound 2A-29-F

[0568] Experimental procedure:

[0569] Compound 2A-29-E (3.00 g, 6.38 mmol), 3301-F (1.68 g, 8.29 mmol, synthesized according to steps one and two of Example 8), TsOH (55 mg, 0.32 mmol), and toluene (60 mL) were added to a 100 mL flask. The mixture was heated under reflux for 3 h to remove water. The reaction solution was concentrated under reduced pressure to obtain crude compound 2A-29-F. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v ratio 1:4) to give 2.30 g of compound 2A-29-F, yield 70.98%.

[0570] Step 7: Synthesize compound 2A-29-G

[0571] Experimental procedure:

[0572] Compound 2A-29-F (3.5 g, 5.34 mmol), Pd / C (0.70 g), and THF (21 mL) were added to a 250 mL flask. The mixture was purged with hydrogen three times and reacted at 25 ± 5 °C for 24 h. Pd / C was removed by filtration to obtain a solution of compound 2A-29-G.

[0573] Step 8: Synthesize compound 2A-29-H

[0574] Experimental procedure:

[0575] Add EDCI (2.54 g, 13.29 mmol), pyridine (1.26 g, 15.93 mmol), DMAP (0.19 g, 1.56 mmol), and DCM (25 mL) to a 100 mL flask. Stir at 25 ± 5 °C for 10 min. Add the solution of compound 2A-29-G obtained in step 7 and BSB (2.76 g, 10.06 mmol, synthesized according to step 3 of Example 1). Stir the reaction at 25 ± 5 °C for 15 h. Add water (50 mL), extract and separate the phases. Extract the aqueous phase with DCM (50 mL), and combine the organic phases. Wash the organic phase with water (50 mL), concentrate under reduced pressure, and obtain crude compound 2A-29-H. Purify the crude product by silica gel column chromatography, eluting with EA and n-heptane (v / v ratio 1:9), to obtain 4.00 g of compound 2A-29-H, with a yield of 75.8%.

[0576] Step 9: Synthesize compound 2A-29-I

[0577] Experimental procedure:

[0578] Compound 2A-29-H (4.00 g, 4.05 mmol), anhydrous ethanol (20 mL), and 50% hydrazine hydrate solution (2.03 g, 20.34 mmol) were added to a 50 mL flask. The mixture was refluxed at 80 ± 5 °C for 2 h. Insoluble matter was removed by filtration, and the filtrate was concentrated under reduced pressure at 45 °C to obtain crude compound 2A-29-I. The crude product was purified by silica gel column chromatography, eluting with DCM and methanol (19:1 v / v) to give 3.10 g of compound 2A-29-I, with a yield of 89.3%.

[0579] Step 10: Synthesize compound 2A-29

[0580] Experimental procedure:

[0581] Compound 2A-29-I (3.1 g, 3.61 mmol), 37% formaldehyde aqueous solution (1.46 g, 18.06 mmol), Pd / C (0.62 g), and anhydrous ethanol (31 mL) were added to a 100 mL flask. The mixture was purged with hydrogen three times and stirred at 25 ± 5 °C for 18 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure at 45 °C to obtain crude compound 2A-29. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (65:35 v / v), to give 180 mg of a colorless, transparent oily product of compound 2A-29, with an HPLC purity of 92.48% and a yield of 5.6%.

[0582] 1 H NMR (600MHz, CDCl3) δ4.93 (t, J = 4.7Hz, 1H), 4.49-4.47 (m, 2H), 4.09-4.07 ( m,4H),3.91-3.88(m,2H),3.59-3.55(m,4H),3.43-3.39(m,4H),2.56-2.36 (m,2H),2.31-2.30(m,4H),2.29-2.27(m,6H),1.89-1.81(m,2H),1.73-1.6 1(m,12H),1.57-1.54(m,11H),1.41-1.16(m,33H),0.89(t,J=6.9Hz,12H).

[0583] LC-MS(ESI):Calculated for 885.73,Found(M+H):887.0.

[0584] Example 11 Synthesis of aminolipid compound 2B-35

[0585] Step 1: Synthesize compound 2B-35-A

[0586] Experimental procedure:

[0587] (1) Heptadecanoic acid ketone was prepared according to CN114805049A.

[0588] (2) EDCI (15.24 g, 79.51 mmol), pyridine (7.55 g, 95.41 mmol), and DCM (100 mL) were added to a 250 mL flask and stirred at 25 ± 5 °C for 10 min. Heptadecanone (10.00 g, 31.80 mmol), DMAP (1.17 g, 9.51 mmol), and benzyl alcohol (7.22 g, 66.79 mmol) were added. The mixture was stirred at 25 ± 5 °C for 3 h. Water (100 mL) was added, and the mixture was extracted and separated, collecting the organic phase. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2B-35-A. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v ratio 1:9) to obtain 10.80 g of compound 2B-35-A, with a yield of 68.7%.

[0589] Step 2: Synthesize compound 2B-35-B

[0590] Experimental procedure:

[0591] Add methyltriphenylphosphine bromide (11.2 g, 31.35 mmol) and anhydrous THF (112 mL) to a 500 mL flask under nitrogen protection. Cool to 0 ± 5 °C and slowly add potassium tert-butoxide solution (potassium tert-butoxide (3.16 g, 28.18 mmol) dissolved in anhydrous THF (34 mL)) dropwise to the reaction. After the addition is complete, continue the reaction at 0 ± 5 °C for 1 h. Maintain the temperature at 0 ± 5 °C and add compound 2B-35-A solution (2B-35-A (10.86 g, 21.95 mmol) dissolved in anhydrous THF (22 mL)) dropwise. After the addition is complete, continue the reaction at 0 ± 5 °C for 2 h. Add water (112 mL), extract and separate the phases. Extract the aqueous phase with DCM (112 mL), combine the organic phases, and concentrate under reduced pressure to obtain crude compound 2B-35-B. The crude product was purified by silica gel column chromatography, eluted with EA and n-heptane (volume ratio 1:9), to give 7.81 g of compound 2B-35-B, yield 72.2%.

[0592] Step 3: Synthesize compound 2B-35-C

[0593] Experimental procedure:

[0594] Compound 2B-35-B (7.80 g, 15.83 mmol), DCM (60 mL), methylrhenium trioxide (79 mg, 0.32 mmol), and 30% hydrogen peroxide (2.64 g, 23.74 mmol) were added to a 500 mL flask. The mixture was purged with nitrogen three times and reacted at 25 ± 5 °C for 16 h. Water (60 mL) was added, and the mixture was extracted and separated. The organic phase was collected and concentrated to dryness under reduced pressure. Acetonitrile (70 mL), water (23 mL), and 3.6 M sulfuric acid solution (11 mL) were added, and the mixture was reacted at 25 ± 5 °C for 2 h. The mixture was concentrated under reduced pressure, and DCM (25 mL) was added to the residue. The mixture was extracted and separated, and the aqueous phase was extracted with DCM (25 mL). The organic phases were combined and concentrated under reduced pressure to obtain crude compound 2B-35-C. The crude product was purified by silica gel column chromatography, eluted with EA and n-heptane (volume ratio 1:4), to give 4.10 g of compound 2B-35-C, yield 49.2%.

[0595] Step 4: Synthesize compound 2B-35-D

[0596] Experimental procedure:

[0597] Compound 2B-35-C (1.75 g, 3.32 mmol), 3,3-dimethoxy-N,N-dimethyl-1-propane (0.78 g, 5.32 mmol), toluene (17.5 mL), and TsOH (0.92 g, 5.32 mmol) were added to a 100 mL flask, and the mixture was refluxed to remove water for 3 h. Saturated sodium bicarbonate solution (17.5 mL) was added, and the mixture was stirred for 5 min. The mixture was then extracted twice with EA (17.5 mL), and the combined organic phases were concentrated under reduced pressure to obtain crude compound 2B-35-D. The crude product was purified by silica gel column chromatography, eluting with methanol and DCM (v / v, 1:19) to give 1.50 g of compound 2B-35-D, in 73.9% yield.

[0598] Step 5: Synthesize compound 2B-35-E

[0599] Experimental procedure:

[0600] Compound 2B-35-D (1.50 g, 2.46 mmol), Pd / C (0.30 g), and THF (9 mL) were added to a 50 mL flask. The mixture was purged with hydrogen three times and reacted at 20 ± 5 °C for 12 h. The mixture was then filtered to obtain a solution of compound 2B-35-E.

[0601] Step 6: Synthesize compound 2B-35

[0602] Experimental procedure:

[0603] Add EDCI (1.17 g, 6.11 mmol), pyridine (0.58 g, 7.33 mmol), DMAP (0.09 g, 0.73 mmol), and DCM (10 mL) to a 50 mL flask. Stir at 25 ± 5 °C for 10 min. Then add the 2B-35-E solution obtained in step 5 and BSB solution (BSB (1.34 g, 4.89 mmol, synthesized according to step 3 of Example 1), dissolved in THF (10 mL)). Stir the reaction mixture at 25 ± 5 °C for 5 h. Concentrate the reaction solution under reduced pressure to obtain crude compound 2B-35. Purify the crude product by silica gel column chromatography, eluting with n-heptane and EA (9:1 v / v), to obtain 500 mg of colorless, transparent, oily compound 2B-35 product with HPLC purity of 91.93% and yield of 21.7%.

[0604] 1 H NMR (600MHz, CDCl3) δ5.04 (t, J = 4.7Hz, 1H), 4.53-4.51 (m, 2H), 4.13-4.11 (m,4H),3.77(d,J=8.1Hz,1H),3.63-3.59(m,5H),3.47-3.43(m,4H),2.50- 2.48(m,2H),2.40(s,1H),2.36-2.27(m,10H),1.95-1.83(m,2H),1.77-1. 69(m,8H),1.66-1.58(m,14H),1.43-1.23(m,41H),0.93(t,J=6.9Hz,12H).

[0605] LC-MS(ESI): Calculated for 941.79, Found(M+H): 943.1.

[0606] Example 12 Synthesis of aminolipid compound 2A-39

[0607] Step 1: Synthesis of compound 2A-39-G

[0608] Experimental procedure:

[0609] 1-Bromo-3,3-dimethoxypropane (5.00 g, 27.32 mmol), 2M dimethylamine solution (2.46 g, 54.63 mmol), potassium carbonate (7.55 g, 54.63 mmol), and acetonitrile (25 mL) were added to a 100 mL flask. The mixture was heated to 65 ± 5 °C and reacted for 24 h. Water (100 mL) and DCM (100 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (100 mL), and the organic phases were combined and concentrated under reduced pressure to give 3.20 g of compound 2A-39-G, with a yield of 79.6%.

[0610] Step 2: Synthesize compound 2A-39-A

[0611] Experimental procedure:

[0612] Octanediol (32.06 g, 219.25 mmol) and DMF (150 mL) were added to a 500 mL flask and stirred at 0 ± 5 °C. 60% sodium hydride (3.87 g, 96.47 mmol) was added in portions under controlled temperature, and the reaction mixture was heated to 25 ± 5 °C for 1 h. The reaction mixture was then cooled to 0 °C, and benzyl bromide (15.00 g, 87.70 mmol) diluted with DMF (7.5 mL) was added dropwise to the reaction mixture. After the addition was complete, the mixture was stirred at 0 ± 5 °C for 20 min. The temperature was then raised to 25 ± 5 °C and the reaction mixture was reacted for 5 h. Water (75 mL) was added dropwise to quench the reaction mixture with stirring. The mixture was extracted twice with EA (45 mL), and the organic phases were combined. The mixture was then washed with water (75 mL) and concentrated under reduced pressure to obtain the crude compound 2A-39-A. The crude compound was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v ratio 1:6). 15.20 g of compound 2A-39-A was obtained, with a yield of 73.3%.

[0613] Step 3: Synthesize compound 2A-39-B

[0614] Experimental procedure:

[0615] Compound 2A-39-A (15.20 g, 64.31 mmol) and DCM (152 mL) were added to a 500 mL flask and stirred until dissolved. Dys-Martin reagent (32.73 g, 77.17 mmol) was added in portions, and the mixture was reacted at 25 ± 5 °C for 4 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain crude compound 2A-39-B. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v, 1:9) to give 13.10 g of compound 2A-39-B, in 86.9% yield.

[0616] Step 4: Synthesize compound 2A-39-C

[0617] Experimental procedure:

[0618] Compound 2A-39-B (13.10 g, 55.90 mmol) was added to a 500 mL flask, followed by 3-benzyl-5-(2-hydroxyethyl)-4-methyl-1,3-thiazolyl chloride (1.81 g, 6.71 mmol) and TEA (17.82 g, 176.09 mmol). The mixture was stirred at 80 ± 5 °C for 3 h. The solution was concentrated under reduced pressure to obtain crude compound 2A-39-C. The crude compound was purified by silica gel column chromatography, eluting with EA and n-heptane (v / v, 1:5) to give 7.30 g of compound 2A-39-C, in 55.7% yield.

[0619] Step 5: Synthesize compound 2A-39-D

[0620] Experimental procedure:

[0621] Compound 2A-39-C (7.30 g, 1.00 eq) and DCM (25 mL) were added to a 500 mL flask. NaBH4 (465 mg, 0.8 eq) was added in portions at -5 ± 5 °C, followed by dropwise addition of methanol (3.7 mL). The reaction was stirred and maintained at this temperature for 2 h. At -5 ± 5 °C, a saturated ammonium chloride solution (25 mL) was added dropwise, and the mixture was extracted and separated. The aqueous phase was extracted with DCM (25 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-39-D. The crude product was purified by silica gel column chromatography, eluting with methanol and DCM (v / v ratio 1:19) to give 7.30 g of compound 2A-39-D, with a yield of 99.6%.

[0622] Step 6: Synthesize compound 2A-39-E

[0623] Experimental procedure:

[0624] Compound 2A-39-D (2.00 g, 4.25 mmol), 3,3-dimethoxy-N,N-dimethyl-1-propylamine (1.00 g, 6.80 mmol), and toluene (20 mL) were added to a 100 mL flask. The mixture was refluxed for 2 h to remove water. TsOH (808 mg, 4.69 mmol) was then added, and the reaction was continued for another 2 h. A saturated potassium bicarbonate solution (20 mL) and EA (20 mL) were added, and the mixture was extracted and separated. The aqueous phase was extracted with EA (20 mL), and the organic phases were combined and concentrated under reduced pressure to obtain crude compound 2A-39-E. The crude product was purified by silica gel column chromatography, eluting with EA and n-heptane (2:1 v / v) to give 1.50 g of compound 2A-39-E, in 63.8% yield.

[0625] Step 7: Synthesize compound 2A-39-F

[0626] Experimental procedure:

[0627] Compound 2A-39-E (1.50 g, 2.71 mmol), Pd / C (0.30 g), and methanol (9 mL) were added to a 50 mL flask. The mixture was purged with hydrogen three times and reacted at 20 ± 5 °C for 24 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give 0.86 g of compound 2A-39-F, with a yield of 85.1%.

[0628] Step 8: Synthesize compound 2A-39

[0629] Experimental procedure:

[0630] Add EDCI (1.03 g, 5.35 mmol), pyridine (0.51 g, 6.42 mmol), DMAP (0.08 g, 0.64 mmol), and DCM (16 mL) to a 50 mL flask. Stir for 10 min, then add BSA (1.24 g, 4.29 mmol, as in step 2 of Example 5) and compound 2A-39-F (0.80 g, 2.14 mmol). React at 20 ± 5 °C for 3 h. Add saturated potassium bicarbonate solution (16 mL), extract and separate, concentrate the organic phase under reduced pressure to obtain crude compound 2A-39. Purify the crude product by silica gel column chromatography, eluting with n-heptane and EA (71:29 v / v), to obtain 122 mg of colorless, transparent oily compound 2A-39 product, HPLC purity 95.25%, yield 6.2%.

[0631] 1 H NMR (600MHz, CDCl3) δ4.94 (t, J = 4.7Hz, 1H), 4.51-4.49 (m, 2H), 4.07-4.04 (m, 4H),4.01-3.88(m,2H),3.59-3.55(m,5H),3.43-3.39(m,4H),2.47-2.44(m,2H ),2.39-2.37(m,4H),2.27(s,6H),1.95-1.91(m,4H),1.85-1.79(m,2H),1.63 -1.61(m,4H),1.59-1.48(m,11H),1.39-1.24(m,40H),0.89(t,J=6.9Hz,12H).

[0632] LC-MS(ESI): Calculated for 913.76, Found(M+H): 915.0.

[0633] Example 13 Synthesis of aminolipid compound 2A-43

[0634] Experimental procedure:

[0635] Compound 2A-43-F was synthesized according to steps one through seven of Example 12, and compound 1-39-E was synthesized according to step five of Example 7. Then, compound 2A-43 was synthesized according to step eight of Example 12, wherein compound 2A-43-F (0.80 g, 2.14 mmol) and compound 1-39-E (1.30 g, 4.28 mmol) were produced, yielding crude compound 2A-43. The crude product was purified by silica gel column chromatography, eluting with n-heptane and EA (3:1 v / v), to give 200 mg of colorless, transparent, oily compound 2A-43 product with an HPLC purity of 97.43% and a yield of 9.9%.

[0636] 1 H NMR (600MHz, CDCl3) δ4.93(t,J=4.8Hz,1H),4.48-4.46(m,2H),4.06-4.04(m,4H),4.00-3.90(m,2H),3.68-3.64(m,5H),3.42-3.38(m,4H) ),2.44-2.40(m,2H),2.34-2.32(m,4H),2.25(s,6H),1.87-1.80(m,2H),1.72-1.49(m,23H),1.39-1.23(m,40H),0.89(t,J=6.9Hz,12H).

[0637] LC-MS(ESI): Calculated for 941.79, Found(M+H): 943.0.

[0638] Experimental Example 1. Preparation of lipid nanoparticles encapsulating luciferase mRNA (Fluc mRNA)

[0639] (1) Ingredients:

[0640] Add the specified amount of Fluc mRNA stock solution and 25mM sodium acetate buffer to a container and mix well to obtain the aqueous phase.

[0641] Representative aminolipid compounds, DSPC, cholesterol (CHO-HP), and DMG-PEG 2000 of this disclosure were dissolved in anhydrous ethanol to prepare solutions with concentrations of 20 mg / mL, 10 mg / mL, 20 mg / mL, and 10 mg / mL, respectively. The molar ratio of aminolipid compound:DSPC:CHO-HP:DMG-PEG 2000 was 48:10:40.5:1.5. The four solutions were then transferred to separate solutions, mixed, and the resulting alcohol phase was prepared.

[0642] (2) Encapsulation: The aqueous and alcohol phases of step (1) were encapsulated using an MPE-L2 microfluidic preparation instrument to obtain lipid nanoparticles encapsulating mRNA.

[0643] (3) Dialysis: The product from step (2) was loaded into a dialysis bag and placed in Tris Buffer-8% (m / V) sucrose solution for displacement to remove residual ethanol, unassembled lipids, and other components. Dialysis was performed for 2 hours at room temperature in the dark with magnetic stirring. The dialysate was changed every hour during the dialysis process.

[0644] (4) The product from step (3) is passed through a 0.22 μm microporous membrane for sterilization, and then dispensed to prepare various LNP formulations encapsulating Fluc mRNA. Each LNP formulation contains different amino lipid compounds, and the particle size of each LNP formulation is 70 nm to 120 nm. The encapsulation rate of Fluc mRNA is over 90%.

[0645] Experimental Example 2. In vivo delivery level test of lipid nanoparticles encapsulating Fluc mRNA in mice

[0646] Female BALB / c mice aged 6–8 weeks were housed in an SPF-grade enclosure. The LNP formulation encapsulated with Fluc mRNA prepared in Example 1 was administered to the mice via tail vein injection, at a dose of 1 mpk per mouse, based on the mRNA content of the LNP formulation. Six hours after administration, the mice were anesthetized with isoflurane inhalation and injected with the luciferase-enhancing substrate D-Luciferin (150 mg / kg). The animals were placed in a supine position, and the distribution and expression intensity of fluorescence signals in the vivo animals and various organs were observed using an IVIS in vivo imaging system. The LNP formulation prepared using control compounds 1 and 2 as aminolipid compounds served as the control LNP formulation. The preparation method of the control LNP formulation was the same as that of the LNP formulation in Example 1, except that the representative aminolipid compounds of this disclosure were replaced with control compounds 1 and 2. The structures of control compounds 1 and 2 are shown below.

[0647] The experimental results are shown in Table 2, Figure 1, and Figures 2A-2F. The unit for the average photon number in Table 2, Figure 1, and Figures 2A-2F is p / s / cm. 2All tested aminolipid compounds exhibited systemic delivery activity, with control compound 2 showing relatively low delivery activity. Most aminolipid compounds showed a significant advantage in spleen delivery activity compared to control compound 1, particularly aminolipid compounds 1-06, 1-09, 1-32, 1-33, and 1-39, which exhibited high spleen / liver ratios, indicating a spleen-preferred delivery pattern.

[0648] Table 2. Fluorescence expression intensity induced by lipid nanoparticles encapsulating Fluc mRNA

[0649] Experimental Example 3. In vivo delivery level test of lipid nanoparticles encapsulating hEPO mRNA in rats

[0650] Following the method described in Example 1, Fluc mRNA was replaced with hEPO mRNA to prepare an LNP formulation encapsulating hEPO mRNA. Female SD rats weighing 180–220 g were selected and housed in an SPF-protected laboratory environment. The prepared hEPO mRNA-encapsulated LNP formulation was administered to the rats via tail vein injection, with each rat receiving a dose of 1 mpk based on the mRNA content in the LNP formulation. Blood was collected from the orbital sinus at 6 h and 24 h after administration, centrifuged, and serum was obtained to detect the expression level of hEPO in the serum.

[0651] The experimental results are shown in Table 3, Figure 3, and Figure 4. PBS buffer solution was used as a blank control, and control compound 2 (an aminolipid compound) was used as the LNP formulation encapsulating hEPO mRNA as the control LNP formulation. The hEPO concentration in Table 3 is expressed in ng / mL. As can be seen from Table 3, Figure 3, and Figure 4, all tested aminolipid compounds exhibited delivery activity, with control compound 2 showing relatively low delivery activity.

[0652] Table 3. Serum hEPO concentrations 6 h and 24 h after drug administration

[0653] It should also be noted that the various specific technical features described in different implementations or embodiments of this disclosure can be combined in any suitable manner without contradiction to provide other implementation methods or embodiments. To avoid unnecessary repetition, this disclosure will not further describe the various possible combinations.

[0654] In addition to those described herein, various modifications to this disclosure will be apparent to those skilled in the art based on the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

Claims

Amino lipid compounds as shown in formula (I), formula (II) or formula (III): Or its pharmaceutically acceptable salt, or its stereoisomer, in, X1 and X2 are independently selected from O or S; R1 is Wherein R' is H, an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C1-C6 heteroalkyl, or an optionally substituted C2-C6 heteroalkenyl; R” is H, an optionally substituted C1-C6 alkyl, an optionally substituted C2-C6 alkenyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted C2-C6 heteroalkenyl, or an optionally substituted C2-C6 heteroalkenyl. X3 and X4 are independently selected from O or S; R M H, halogen, -OH, -R * -N(R) * )2, -CN, -N3, -C(=O)OH, -C(=O)OR * -OC(=O)R * -OR * -SR * -S(=O)R * -S(=O)OR * -S(=O)2OR * -NO2, -S(=O)2N(R) * )2、-N(R * )S(=O)2R * -NH(CH2) t1 N(R * 2、-NH(CH2) p1 O(CH2) q1 N(R * 2、-NH(CH2) s1 OR * -N((CH2) s1 OR * )2、-N(R * -Carbon ring, -N(R) * - Heterocyclic rings, -N(R) * -Aryl, -N(R) * )-Heteroaryl, -N(R * (CH2) t1 -Carbon ring, -N(R) * (CH2) t1 -heterocyclic, -N(R) * (CH2) t1 -Aryl, -N(R) * (CH2) t1 - Heteroaryl, carbocyclic, heterocyclic, aryl or heteroaryl; Each R * It is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Each t1 is independently 1, 2, 3, 4 or 5; p1 is 1, 2, 3, 4 or 5; q1 is 1, 2, 3, 4, or 5; Each s1 is independently 1, 2, 3, 4 or 5; R N It is H or C1-C3 alkyl; r is 0 or 1; n is any integer from 0 to 10; L3 is a bond, an optional substituted C1-C6 alkylene group, or an optional substituted C1-C6 heteroalkylene group; R2 is H, a C1-C6 hydrocarbon group, or a C1-C6 heterohydrocarbon group; L1 and L2 are independently selected from bonded, optionally substituted C1-C3 alkylene or optionally substituted C2-C3 alkenyl groups; A1 and A2 are independently selected from C1 to C2. 10 Alkylene or C1-C 10 Heteroalkyl; A3 and A4 are independently selected from C1-C6 alkylene, C1-C6 heteroalkylene, or bonded; R3 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5); R4 is C1~C 24 Hydrocarbon group, C1-C containing O or S 24 heteroalkyl groups, -M1Y, -YM a -M2YM b -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6); Each M1 is independently C1~C 24 Hydroxyl group; Each M2 is independently C1 to C 12 Hydroxyl group; Each M a Independently C1~C 24 hydrocarbon group; Each M b Independently C1~C 15 hydrocarbon group; Z1 and Z2 are independently selected from -C(=O)O- or -OC(=O)-; Each R a It is independently an H or C1-C8 hydrocarbon group; Each R5 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ; Each R6 is independently C3 to C 12 Hydrocarbon group, C3~C 12 heterohydrocarbon groups, -M3Y, -YM c or -M4YM d ; Each M3 is independently C3~C 12 Hydroxyl group; Each M4 is independently a C1–C8 hydrocarbon group; Each M c Independently C3~C 12 hydrocarbon group; Each M d Independently composed of C1–C8 hydrocarbon groups; Each Y is an independent 3-7 member carbon ring. The aminolipid compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein X1 and X2 are O. The aminolipid compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z1 and Z2 are -C(=O)O-. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (I), wherein R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is -C(R a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6); Preferably, R3 is C1 to C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is -C(R a (OR6)2; or Preferably, R3 is C1 to C 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; or Preferably, R3 is C1 to C 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (I), wherein R3 is -CH(OR5)2 and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (I), wherein X1 and X2 are O, and R3 is C1 to C2. 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; Preferably, X1 and X2 are 0, and R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (I), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (I), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2; or Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound has a structure as shown in formula (IV): The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (II), wherein R3 is C1 to C2. 24 hydrocarbon group, -C(R) a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is -C(R a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6); Preferably, R3 is C1 to C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is -C(R a (OR6)2; or Preferably, R3 is C1 to C 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; or Preferably, R3 is C1 to C 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (II), wherein R3 is -C(R a (OR5)2、-C(R) a (SR5)2 or -C(R) a (SR5)(OR5), R4 is C1~C 24 hydrocarbon group, -C(R) a (OR6)2、-C(R) a (SR6)2 or -C(R) a (SR6)(OR6); Preferably, R3 is -C(R a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a (OR6)2; or Preferably, R3 is -CH(OR5)2, and R4 is Cl~C 24 Hydrocarbon group or -CH(OR6)2; or Preferably, R3 is -CH(OR5)2, and R4 is Cl~C 24 Hydrocarbon group. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (II), wherein R3 is -CH(OR5)2 and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (II), wherein X1 and X2 are O, and R3 is C1 to C2. 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; Preferably, X1 and X2 are 0, and R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (II), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group or -CH(OR6)2; Preferably, X1 and X2 are O, R3 is -CH(OR5)2, and R4 is Cl~C 24 Hydrocarbon group. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (II), wherein X1 and X2 are O, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (II), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is -CH(OR6)2; Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (II), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is C1~C 24 Hydrocarbon group or -CH(OR6)2; Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is C1~C 24 hydrocarbon group; or Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound has a structure as shown in formula (V): The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein R3 is C1 to C2. 24 Hydrocarbon group, R4 is C1 to C2. 24 Hydrocarbon group. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (III), wherein Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a (OR6)2; Preferably, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2; or Preferably, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (III), wherein Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (III), and Where R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The amino lipid compound is shown in formula (III), wherein R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 hydrocarbon group or -C(R) a (OR5)2, R4 is C1~C 24 hydrocarbon group or -C(R) a (OR6)2; Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group or -CH(OR5)2, R4 is C1 to C2. 24 Hydrocarbon group or -CH(OR6)2; or Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2; or Preferably, X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, and R4 is -CH(OR6)2. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound has a structure as shown in formula (VI): The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein X1 and X2 are O, and R3 is C1 to C2. 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein X1 and X2 are O, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, and R3 is C1~C 24 Hydrocarbon group, R4 is -CH(OR6)2, A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The aminolipid compound is shown in formula (III), wherein X1 and X2 are O, Z1 and Z2 are -C(=O)O-, R3 is -CH(OR5)2, R4 is -CH(OR6)2, and A1 or A2 is C2~C 10 Alkylene, C2-C 10 alkenyl or C1-C 10 Heteroalkyl groups. The aminolipid compound according to any one of claims 1 to 29, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, L1 and L2 are independently selected from C1 to C3 alkylene groups that are bonded or optionally substituted, and the ring containing L1 and L2 is a 4-7 membered heterocycle; Preferably, L1 and L2 are independently selected from C1-C3 alkylene groups that are either bonded or unsubstituted; Preferably, the heterocycle is a 5- or 7-membered heterocycle, more preferably a 5- or 6-membered heterocycle. The aminolipid compound according to any one of claims 1 to 30, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, L1 and L2 are not both keys; or L1 and L2 are not simultaneously unsubstituted C1 alkylene groups; or L1 and L2 are both bonds. The aminolipid compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein: R' is H, an optionally substituted C1-C6 alkyl group, or an optionally substituted C2-C6 alkenyl group; and R” is H, an optionally substituted C1-C6 alkyl group, an optionally substituted C2-C6 alkenyl group, or The aminolipid compound according to any one of claims 1 to 32, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R' and R” are independently selected from optional substituted C1 to C3 alkyl groups; Preferably, R' and R" are independently selected from unsubstituted C1 to C3 alkyl groups. The aminolipid compound according to any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, X3 and X4 are O, R M -N(R) * )2, r is 1; Preferably, each R * Independently, it is H or a C1-C3 alkyl group; Preferably, one of the R * For H. The aminolipid compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R' is an optionally substituted C1-C3 alkyl group, and R” is Where R * It is a C1 to C3 alkyl group; Preferably, R' is an unsubstituted C1-C3 alkyl group; more preferably, R' is methyl or ethyl. Preferably, R * It can be methyl or ethyl. The aminolipid compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R N For H. The aminolipid compound according to any one of claims 1 to 36, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, n is any integer from 1 to 5. The aminolipid compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, L3 is a C1-C6 alkylene group that is either bonded or optionally substituted; Preferably, L3 is an unsubstituted C1-C4 alkylene group. The aminolipid compound according to any one of claims 1 to 38, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R2 is H or a C1-C6 alkyl group; Preferably, R2 is H. The aminolipid compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R3 is C1~C 24 Alkyl or C2-C 24 alkenyl; Preferably, R3 is a straight-chain C3~C 15 Alkyl group; or, R3 is a branched C8-C12 group. 24 alkyl. The aminolipid compound according to any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R4 is C1~C 24 Alkyl or C2-C 24 alkenyl; Preferably, R4 is a straight-chain C3~C 15 Alkyl group; or, R4 is a branched C8-C10 group. 24 alkyl. The aminolipid compound according to any one of claims 1 to 41, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, Each R5 is independently C3 to C 12 Alkyl or C3-C 12 alkenyl; Preferably, each R5 is independently C3 to C5. 12 Alkyl; or Preferably, each R5 is independently a C3 to C9 alkyl group; or Preferably, each R5 is independently a C5-C8 alkyl group; or Preferably, each R5 is independently a straight-chain C5-C8 alkyl group. The aminolipid compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, R6 is independently C3~C 12 Alkyl or C3-C 12 alkenyl; Preferably, each R6 is independently C3 to C6. 12 Alkyl; or Preferably, each R6 is independently a C3 to C9 alkyl group; or Preferably, each R6 is independently a C5-C8 alkyl group; or Preferably, each R6 is independently a straight-chain C5-C8 alkyl group. The aminolipid compound according to any one of claims 1 to 43, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, A1 and A2 are independently C3 to C 10 Alkylene; Preferably, A1 and A2 are independently C5 to C8 alkylene groups. The aminolipid compound according to any one of claims 1 to 44, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, A3 and A4 are independently C1-C6 alkylene groups; Preferably, A3 and A4 are independently C2-C4 alkylene groups. The aminolipid compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the aminolipid compound is selected from one of the following structures: Empty lipid nanoparticles containing any of the amino lipid compounds according to claims 1 to 46. The air-filled lipid nanoparticles according to claim 47, wherein, The empty lipid nanoparticles also contain one or more of the following: auxiliary lipids, structural lipids, and PEG-lipids (polyethylene glycol-lipids); Preferably, the empty lipid nanoparticles further contain the auxiliary lipids, the structural lipids, and the PEG-lipids. Lipid-loaded nanoparticles comprising the empty lipid nanoparticles as described in claim 47 or 48 and the active ingredient. The lipid-loaded nanoparticles according to claim 49, wherein, The active ingredient is a pharmaceutical active ingredient; The active pharmaceutical ingredient is preferably a bioactive ingredient, which is preferably a nucleic acid. The nucleic acid is preferably selected from one or more of RNA, antisense oligonucleotides, and DNA. The RNA is preferably selected from messenger RNA (mRNA), ribosomal RNA (rRNA), microRNA (miRNA), transfer RNA (tRNA), small interfering RNA (siRNA), small nuclear RNA (snRNA), small hairpin RNA (shRNA), single-stranded guide RNA (sgRNA), Cas9 mRNA, or mixtures thereof. The DNA is preferably a plasmid. A pharmaceutical composition comprising the lipid-loaded nanoparticles as described in claim 49 or 50, and a pharmaceutically acceptable carrier, diluent, or excipient. Use of the empty lipid nanoparticles of claim 47 or 48 in the preparation of a delivery carrier; preferably, the delivery carrier is used to deliver an active ingredient. Methods for delivering active ingredients to cells, tissues, or organs include: The lipid-loaded nanoparticles of claim 49 or 50 are brought into contact with the cells, tissues or organs. Methods for generating polypeptides and / or proteins of interest in mammalian cells, including: Provide the lipid-loaded nanoparticles of claim 49 or 50 containing mRNA, and contact the cells with the lipid-loaded nanoparticles. Use of the lipid-loaded nanoparticles of claim 49 or 50 or the pharmaceutical composition of claim 51 in the treatment and / or prevention of diseases or conditions. Methods for treating and / or preventing diseases or conditions in mammals in need, including: Administer to mammals a therapeutic and / or preventative amount of the lipid-loaded nanoparticles of claim 49 or 50 or the pharmaceutical composition of claim 51. Use of the lipid-loaded nanoparticles of claim 49 or 50 or the pharmaceutical composition of claim 51 in the preparation of a medicament; Preferably, the drug is used to treat and / or prevent diseases or conditions; or Preferably, the drug is used for gene therapy, protein replacement therapy, antisense therapy, or treatment via interfering RNA, as well as gene vaccination, wherein the gene vaccination is preferably used for the treatment and / or prevention of cancer, allergies, toxicity, and pathogen infection. The lipid-loaded nanoparticles of claim 49 or 50 or the pharmaceutical composition of claim 51 are used as a drug; Preferably, the drug is used to treat and / or prevent diseases or conditions; or Preferably, the drug is used for gene therapy, protein replacement therapy, antisense therapy, or treatment via interfering RNA, as well as gene vaccination, wherein the gene vaccination is preferably used for the treatment and / or prevention of cancer, allergies, toxicity, and pathogen infection. A medicine comprising the lipid-loaded nanoparticles of claim 49 or 50 or the pharmaceutical composition of claim 51; Preferably, the drug is used to treat and / or prevent diseases or conditions; or Preferably, the drug is used for gene therapy, protein replacement therapy, antisense therapy, or treatment via interfering RNA, as well as gene vaccination, wherein the gene vaccination is preferably used for the treatment and / or prevention of cancer, allergies, toxicity, and pathogen infection.

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