New cationic lipid compound, and preparation method therefor, composition thereof and use thereof

Abstract

Provided are a new cationic lipid compound, and a preparation method therefor, a composition thereof and the use thereof. The cationic lipid has a structure as represented by formula I. Lipid nanoparticles prepared from the cationic lipid have a stable nanostructure, have a relatively narrow size distribution, and can be stored for a relatively long time at a low temperature. Moreover, the lipid nanoparticles have relatively good biocompatibility and a relatively high in-vivo mRNA transfection efficiency. The core structures of the cationic lipids can all be constructed by means of a Ugi reaction. The reaction involves simple operation and mild reaction conditions, does not require additional catalysts, has high atomic economy, is simple and easy to carry out, and incorporates inexpensive and readily available raw materials, such that not only is higher safety provided, but the industrial production and quality control of the cationic lipids are also facilitated. The new cationic lipid has good application prospects.

Description

Novel cationic lipid compound, preparation method, composition and application thereof

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of Chinese application No. 202311682718.9, filed on December 8, 2023. Said application No. 202311682718.9 is hereby incorporated by reference in its entirety. Technical Field

[0003] The present invention provides a novel cationic lipid molecule and a method for its efficient preparation. The cationic lipid can be used in combination with other lipid components (such as neutral lipids, steroids, and polymer-conjugated lipids) to form a nucleic acid-lipid nanoparticle composition for delivering one or more therapeutic and / or prophylactic agents to mammalian cells or organs and / or producing polypeptides in mammalian cells or organs. In addition to the novel cationic lipid, the lipid nanoparticle composition of the present invention can also include one or more cationic and / or ionizable amino lipids, neutral lipids including polyunsaturated lipids, polymer-conjugated lipids, steroids, and / or therapeutic and / or prophylactic agents in specific ratios. Background Art

[0004] The effective targeted delivery of bioactive substances such as small molecule drugs, proteins and nucleic acids is a long-term medical problem that needs to be overcome. Specifically, it is very difficult to effectively deliver therapeutic nucleic acids (mRNA, DNA, sgRNA, antisense oligonucleotides, nucleic acid aptamers, plasmids, immunostimulatory nucleic acids, etc.) to cells because these species are relatively unstable and have low cell permeability. Therefore, it is necessary to develop methods and compositions that help deliver therapeutic agents and / or prophylactic agents such as nucleic acids into cells or animals.

[0005] Studies have shown that nanoparticle compositions, liposomes, and liposome complexes containing lipid molecules can be used as transport vehicles to effectively deliver biologically active substances such as small molecule drugs, proteins, and nucleic acids to cells and / or intracellular compartments. These compositions generally contain one or more cationic lipids, neutral lipids including polyunsaturated lipids (such as phospholipids), structural lipids (such as steroids), and / or lipids containing polyethylene glycol (polymer-conjugated lipids). Cationic lipids include, for example, amine-containing lipids that can be easily protonated.

[0006] Currently, therapeutic nucleic acids face two problems when used in therapeutic settings. First, free nucleic acids are easily digested and degraded by nucleases in plasma; second, the ability of free nucleic acids to enter intracellular compartments with relevant translation machinery is limited. Lipid nanoparticles (LNPs) formed by cationic lipids and other lipid components (such as neutral lipids, cholesterol, PEGylated lipids and nucleic acids) have been used to prevent the degradation of nucleic acids in plasma and promote the cellular uptake of nucleic acids. In addition, the production and quality control of cationic lipid molecules are also extremely critical aspects. It is necessary and important to construct cationic lipids quickly and efficiently to achieve controllable mass production.

[0007] Therefore, there is still a need to improve cationic lipids and lipid nanoparticles for delivering nucleic acids, so that the improved lipid nanoparticles have more optimized drug delivery performance, prevent nucleic acids from being degraded and cleared in plasma, so as to be suitable for systemic or local delivery, and provide intracellular delivery of nucleic acids; at the same time, improve new cationic lipid structures and production methods to achieve efficient and controllable production and preparation plans and improve safety.

[0008] In summary, there is still a need in this field to develop cationic liposome molecules with completely new structures and efficient and controllable synthesis methods thereof. Summary of the Invention

[0009] The purpose of the present invention is to provide a cationic lipid molecule with a completely new structure and an efficient synthesis method involving these compounds. The cationic lipid molecule of the present invention has the structure shown in Formula I.

[0010] Therefore, in the first aspect, the present invention relates to a compound as shown in Formula I, or an isomer, a pharmaceutically acceptable salt, or a prodrug thereof;

[0011] in:

[0012] Side chain part-W1-L1-(R1) n1 、-W2-L2-(R2) n2 and-W3-L3-(R3) n3 The main chain (i.e., the longest chain) of each independently has 1-40 carbon atoms;

[0013] W1, W2 and W3 are each independently C1-C14 alkylene, C2-C14 alkenylene or C2-C14 alkynylene;

[0014] n1, n2, and n3 are each independently 0, 1, or 2;

[0015] L1, L2 and L3 are each independently selected from the following structures:

[0016] R1, R2 and R3 are each independently H, aryl, cholesteryl, adamantyl, C6-24 alkyl, C6-24 alkenyl, C6-24 alkynyl, C4-C10 cycloalkyl, C4-C10 cycloalkenyl, and any of the alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl groups are connected by one or more ester bonds, amide bonds, carbonate bonds, amino acid ester bonds or ether bonds;

[0017] R1, R2 and R3 can also be selected from 4-10 membered heterocyclic rings, which include one or more heteroatoms selected from N, O, S and Se;

[0018] R4 and R5 are each independently C1-C12 alkyl, C2-C12 alkenyl or C2-C12 alkynyl, wherein the alkyl, alkenyl or alkynyl is arbitrarily substituted with one or more OH, SH, or C1-C4 alkoxy groups, or R4 and R5 are combined with each other to form a 4- to 10-membered heterocyclic ring, which optionally includes one or more heteroatoms of N, O, or S in addition to the N atom connected to M;

[0019] M is a C1-C12 alkylene group, a C2-C12 alkenylene group or a C2-C12 alkynylene group.

[0020] In another preferred embodiment, the compound of formula I has the structure shown in the following formula II:

[0021] X, Y and Z are each independently CH2, NH, S, Se;

[0022] “-----” indicates a single bond or a double bond;

[0023] m and n are each independently 0, 1, 2, 3 or 4;

[0024] The remaining groups are as defined herein.

[0025] Preferably, X and Y are S, Z is CH2, "-----" represents a single bond, and n is 1.

[0026] In another preferred embodiment, the compound has the structure shown in Formula I and the compound has the structure shown in Formula III below:

[0027] The definitions of the groups are as described herein.

[0028] In another preferred embodiment, the side chain portion -W1-L1-(R1) n1 、-W2-L2-(R2) n2 and-W3-L3-(R3) n3The backbone (i.e., longest chain) of each independently has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 carbon atoms. Preferably, it has 6-24 carbon atoms, more preferably 12-22 carbon atoms.

[0029] In another preferred embodiment, W1, W2 and W3 are each independently a C3-C8 alkyl, C3-C8 alkenyl or C3-C8 alkynyl. Preferably, W1, W2 and W3 are each independently a C3-C8 alkyl, C3-C8 alkenyl or C3-C8 alkynyl.

[0030] In another preferred embodiment, a is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. When a is 0, it means that the corresponding L1, L2, and L3 are a bond. Preferably, L1, L2, and L3 are each independently selected from the following structures:

[0031] In another preferred embodiment, R1, R2 and R3 are each independently H, C3-C22 alkyl, C3-C22 alkenyl, C3-C22 alkynyl, C5-C8 cycloalkyl, or C5-C8 cycloalkenyl. Preferably, R1, R2 and R3 are each independently H, C6-C22 alkyl, C6-C22 alkenyl, or C6-C22 alkynyl, more preferably C6-C14 alkyl, C6-C14 alkenyl, or C6-C14 alkynyl.

[0032] In another preferred embodiment, the side chain moieties -W1-L1-R1 and -W2-L2-R2 are each independently selected from the following structures:

[0033] In another preferred embodiment, R4 and R5 are each independently a C1-C6 alkyl, a C2-C6 alkenyl or a C2-C6 alkynyl, and the alkyl, alkenyl or alkynyl is arbitrarily substituted with one or more OH, C1-C4 alkoxy groups, or R4 and R5 are combined with each other to form a 5- to 8-membered heterocyclic ring, and in addition to the N atom connected to M, the heterocyclic ring optionally includes one or more heteroatoms selected from N, O, and S. Preferably, R4 and R5 are each independently a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 2-hydroxyethyl, 3-hydroxypropyl, or R4 and R5 are combined with each other to form one of the following structures:

[0034] Wherein, R is selected from one of C(Ra)2, NRb, O, and S;

[0035] Ra are each independently H, OH, C1-12 alkyl;

[0036] Rb are each independently H, C1-12 alkyl, C(O)C1-12 alkyl;

[0037] a1 is 1, 2, 3, 4, 5, 6, 7 or 8; a2 is 1, 2, 3, 4, 5 or 6; a3 is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. In another preferred embodiment, R4 and R5 are combined to form tetrahydropyrrol-1-yl, piperidinyl-1-yl, piperazin-1-yl, 4-methylpiperazin-1-yl. In another preferred embodiment, M is C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene. Preferably, M is -(CH2) s -, s is 1, 2, 3, 4, 5, or 6.

[0038] In another preferred embodiment, the -MN(R4)(R5) has one of the following structures:

[0039] In another preferred embodiment, the compound is selected from the following group:

[0040] Table 1 Representative compounds

[0041] Unless otherwise defined, the terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs.The terms used in the specification are for describing particular embodiments only and are not intended to limit the invention.

[0042] In this article, Indicates the attachment site.

[0043] As used herein, the term "alkyl" refers to a linear or branched fully saturated hydrocarbon group, preferably a C1-C32 alkyl group, more preferably a C1-C24 alkyl group, and specifically includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, and C32 alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl groups.

[0044] As used herein, the term "alkylene" refers to a group formed by removing a hydrogen atom from an alkyl group.

[0045] As used herein, the term "alkenyl" refers to a straight chain, branched hydrocarbon group having at least one (e.g., 1, 2, 3, 4, 5 or more) carbon-carbon double bond, and includes groups with "cis" and "trans" orientations, or "E" and "Z" orientations. The carbon-carbon double bonds in the alkenyl group can be conjugated or non-conjugated. In some embodiments, C2-C32 alkenyl groups, more preferably C2-C24 alkenyl groups, specifically include C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32 alkenyl groups. Examples of the alkenyl group include ethenyl, propenyl, 1-butenyl, 2-butenyl, 1-pentenyl, 2-pentenyl, and the like, but are not limited thereto.

[0046] As used herein, the term "alkenylene" refers to an alkenyl group by removing a hydrogen atom.

[0047] As used herein, the term "alkynyl" refers to a straight chain or branched hydrocarbon radical having at least one (e.g., 1, 2, 3, 4, 5, or more) carbon-carbon triple bond. The carbon-carbon triple bond in the alkynyl radical can be conjugated or non-conjugated. In some embodiments, preferably C2-C32 alkynyl, more preferably C2-C24 alkynyl, specifically include C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32 alkynyl. The example of alkynyl includes ethynyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, etc., but is not limited thereto.

[0048] As used herein, the term "alkynylene" refers to an alkynyl group by removing a hydrogen atom.

[0049] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic fully saturated hydrocarbon group, preferably a C3-C15 cycloalkyl group, more preferably a C4-C10 cycloalkyl group, and specifically includes C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, and C15 cycloalkyl groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, and adamantyl groups.

[0050] Herein, term " cycloalkenyl " refers to the monocyclic or polycyclic hydrocarbon radical with at least one (such as 1, 2, 3, 4, 5 or more) carbon-carbon double bond, and the carbon-carbon double bond in alkenyl can be conjugated or non-conjugated.In some embodiments, preferably C3-C15 cycloalkenyl, more preferably C4-C15 cycloalkenyl, specifically include C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15 cycloalkenyl.The example of cycloalkenyl includes cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl etc., but is not limited to this.

[0051] Herein, "the alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl groups are arbitrarily connected by one or more ester bonds, amide bonds, carbonate bonds, amino acid ester bonds or ether bonds" means that an ester bond, amide bond, carbonate bond, amino acid ester bond or ether bond can be inserted between any two carbon atoms of the group.

[0052] As used herein, the term "aryl" refers to an aromatic cyclic hydrocarbon group, which may be a monocyclic or condensed ring, preferably a C6-C30, more preferably a C6-C25, C6-C15, or C6-C10 aryl group, specifically including C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, and C30 aryl groups. Examples of aryl groups include phenyl, naphthyl, anthracenyl, phenanthrenyl, and the like.

[0053] As used herein, the term "heterocyclyl" refers to a cyclic group comprising at least one heteroatom selected from nitrogen, oxygen, sulfur, and phosphorus as a ring member; the number of heteroatoms may be 1, 2, 3, 4, 5 or more, wherein the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heterocyclyl may be a monocyclic or fused ring and may have 5 to 25 ring atoms, preferably 5 to 13 ring atoms, and may specifically include 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 ring atoms. The heterocyclyl may be fully saturated, partially saturated, or fully unsaturated, and in the case of partial saturation, may include 1, 2, or 3 carbon-carbon double bonds or carbon-nitrogen double bonds. The heterocyclyl may be attached to the remainder of the molecule via a carbon atom or a heteroatom. Examples of heterocycloalkyl groups include tetrahydropyrrolyl, tetrahydrofuranyl, tetrahydrothiophenyl, dioxolane, dithiolanyl, morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, homopiperidinyl, homopiperazinyl, dihydropyrrolyl, dihydrofuranyl, dihydrothiophenyl, dihydropyridinyl, tetrahydropyridinyl, dihydropiperidinyl, tetrahydropiperidinyl, pyrrolyl, furanyl, thiophenyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl (e.g., 2-pyridinyl), pyridin ... The present invention also includes, but is not limited to, benzopyridinyl, benzothiophene, benzoxazolyl, benzothiazolyl, carbazolyl (including carbazolyl-9-yl), azacarbazolyl (including azacarbazolyl-9-yl), quinolinyl, isoquinolinyl, indolizinyl, azaindolizinyl, dibenzofuranyl, dibenzothiophene, naphthyridinyl, etc., but are not limited to them.

[0054] As used herein, the term "halogen" refers to F, Cl, Br, and I.

[0055] As used herein, the term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of a compound derived from various organic counterions and inorganic counterions known in the art. Pharmaceutically acceptable salts include metal (inorganic) salts and organic salts, including but not limited to those listed in Remington's Pharmaceutical Sciences, 17th edition, page 1418 (1985). Pharmaceutically acceptable salts include, by way of example only, salts of inorganic acids such as hydrochlorides, sulfates, phosphates, diphosphates, hydrobromides, and nitrates: or salts of inorganic acids such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, p-toluenesulfonate, salicylate, and stearate. Similarly, pharmaceutically acceptable cations include but are not limited to sodium, potassium, calcium, aluminum, lithium, and ammonium (especially ammonium salts with secondary amines). Specific salts of the present invention for the reasons cited above include potassium salts, sodium salts, calcium salts, and ammonium salts. In a second aspect, the present invention provides a lipid nanoparticle composition comprising a lipid, and a therapeutic and / or prophylactic agent.

[0056] In another preferred embodiment, the lipid includes a cationic lipid, which is a compound of the structure shown in Formula I, or one or more of its isomers, pharmaceutically acceptable salts, and prodrugs.

[0057] In another preferred embodiment, the present invention provides a lipid nanoparticle composition comprising a compound having a structure shown in Formula I, or one or more of its isomers, pharmaceutically acceptable salts, and prodrugs and a therapeutic agent and / or preventive agent.

[0058] In another preferred embodiment, the lipid further comprises one or more of a neutral lipid, a steroid and a polymer-conjugated lipid.

[0059] In another preferred embodiment, the lipid nanoparticle composition is mainly composed of cationic lipids, neutral lipids, steroids and polymer-conjugated lipids in combination with therapeutic agents and / or preventive agents.

[0060] In another preferred embodiment, the lipid nanoparticle composition is mainly composed of cationic lipids, neutral lipids, steroids and polymer-conjugated lipids in a certain molar ratio or mass ratio in combination with therapeutic agents and / or preventive agents.

[0061] In another preferred embodiment, the compound of the present invention is preferably compound 5, 7 or 8.

[0062] In another preferred embodiment, the neutral lipid is selected from one or more of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and sphingomyelin (SM). In some embodiments, the preferred neutral lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).

[0063] In another preferred embodiment, the steroid is selected from one or more of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, and α-tocopherol. In some embodiments, the preferred steroid is cholesterol.

[0064] In another preferred embodiment, the polymer-conjugated lipid is selected from a PEGylated lipid. The PEGylated lipid is selected from one or more of distearoylphosphatidylethanolamine polyethylene glycol 2000 (DSPE-PEG2000), dimyristoylglycerol-3-methoxypolyethylene glycol 2000 (DMG-PEG2000), and methoxypolyethylene glycol ditetradecyl acetamide (ALC-0159). In some embodiments, the PEGylated lipid is preferably dimyristoylglycerol-3-methoxypolyethylene glycol 2000 (DMG-PEG2000).

[0065] In another preferred embodiment, the therapeutic and / or prophylactic agent comprises a nucleic acid. Preferably, the nucleic acid is RNA selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA, tRNA, mRNA, and sgRNA. In some embodiments, the RNA is selected from mRNA. Preferably, the nucleic acid is used to treat a disease by targeting a specific target; optionally, the nucleic acid molecule can be used to treat cancer, inflammation, fibrotic diseases, autoimmune diseases, infections, congenital and hereditary diseases, connective tissue diseases, digestive system diseases, endocrine diseases, eye diseases, reproductive diseases, cardiovascular diseases, renal and urinary diseases, respiratory diseases, metabolic disorders, musculoskeletal diseases, nervous system diseases, and hematological diseases.

[0066] In another preferred embodiment, the molar ratio of the cationic lipid, neutral lipid, steroid, and polymer-conjugated lipid in the lipid nanoparticle composition is (25-75):(5-25):(15-65):(0.5-10), preferably (40-60):(5-15):(35-45):(1-2). In some embodiments, the optimal ratio is 50:10:38.5:1.5.

[0067] The lipid nanoparticle composition may include one or more therapeutic and / or prophylactic agents, wherein the cationic lipid and the therapeutic and / or prophylactic agent have a mass ratio within a certain range. In some embodiments, the mass ratio of the cationic lipid to the therapeutic and / or prophylactic agent is 10:1 to 40:1, for example, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, or 40:1. The preferred mass ratio for cell administration is 10 to 20:1, more preferably 15:1, and the preferred mass ratio for animal administration is 25 to 35:1, more preferably 30:1.

[0068] The lipid nanoparticles are prepared by mixing an organic phase containing lipid molecules with a buffer phase containing a therapeutic and / or prophylactic agent via rapid mixing, vortexing, or microfluidics. The organic phase is preferably an ethanol phase. The volume ratio of the organic phase to the buffer phase is 1:1-10, preferably 1:2-4, and more preferably 1:3.

[0069] The particle size of the lipid nanoparticles ranges from 50 to 400 nm, preferably from 100 to 300 nm, and the polydispersity index (PDI) ranges from 0.001 to 0.3, preferably from 0.05 to 0.2.

[0070] In a third aspect, the present invention provides use of the compound or lipid nanoparticle composition of the present invention in preparing a delivery system.

[0071] In another preferred embodiment, the delivery system is used to deliver a therapeutic agent and / or a preventive agent into a cell. Preferably, the delivery can be performed in vitro or in vivo.

[0072] In another preferred embodiment, the cell is an animal cell. The cell is selected from the group consisting of endothelial cells, epithelial cells and tumor cells.

[0073] In another preferred embodiment, the in vivo refers to an animal body. Preferably, the cell is in a mammal. More preferably, the mammal is a human.

[0074] The present invention also provides the use of the compound or lipid nanoparticle composition of the present invention for delivering therapeutic and / or prophylactic agents to cells in vitro or in vivo.

[0075] The present invention also provides a method for delivering a therapeutic agent or prophylactic agent to a cell in vitro or in vivo, the method comprising administering a compound of the present invention or lipid nanoparticle composition to the cell. Preferably, the method comprises contacting the compound of the present invention or lipid nanoparticle composition with the cell for a period of time sufficient to allow intracellular delivery to occur.

[0076] In another preferred embodiment, the compound of the present invention or lipid nanoparticle composition is administered by one of the following routes of administration: oral, intranasal, intravenous, intraperitoneal, intramuscular, intraarticular, intralesional, intratracheal, subcutaneous, and intradermal. In a specific embodiment, the compound of the present invention or lipid nanoparticle composition is systemically administered, for example, via an enteral or parenteral route of administration.

[0077] The dosage of the lipid nanoparticle composition of the present invention will depend on the ratio of therapeutic and / or prophylactic agent to lipid and the opinion of the administering physician based on the age, weight and condition of the patient.

[0078] In a fourth aspect, the present invention provides use of the compound or lipid nanoparticle composition of the present invention in the preparation of a medicament for treating a disease or condition in a patient.

[0079] The present invention also provides a method for treating a disease or condition in a patient, comprising administering to the patient a therapeutically effective amount of the lipid nanoparticle composition of the present invention.

[0080] In another preferred embodiment, the diseases or conditions include cancer, inflammation, fibrotic diseases, autoimmune diseases, infections, congenital and hereditary diseases, connective tissue diseases, digestive system diseases, endocrine diseases, eye diseases, reproductive diseases, cardiovascular diseases, kidney and urinary diseases, respiratory diseases, metabolic disorders, musculoskeletal diseases, nervous system diseases and blood system diseases.

[0081] General Synthesis Methods for Cationic Lipid Compounds:

[0082] The cationic lipids of the present invention can all be synthesized by the Ugi four-component reaction "one-pot method", specifically, an aldehyde (A), an amine (B), a carboxylic acid (C) and an isonitrile (D) compound are reacted in a molar ratio of 1:1:1:1. Among them, the aldehyde compound can be purchased from a commercial company or a primary alcohol that can be synthesized by commercial or simple methods is obtained by oxidation of pyridinium chlorochromate (PCC); the isonitrile compound can be purchased from a commercial company or a primary amine that can be synthesized by commercial or simple methods is reacted with ethyl formate to obtain a formamide compound, and the formamide compound is then dehydrated to obtain the formamide compound; the amine and carboxylic acid compounds can both be purchased from commercial companies:

[0083] Therefore, the present invention also provides a method for synthesizing the cationic lipid of the present invention, comprising:

[0084] An aldehyde (A), an amine (B), a carboxylic acid (C) and an isonitrile (D) are reacted to produce a compound represented by formula I;

[0085] wherein R1-R3, W1-W3, L1-L3, M, and n1-n3 are as defined herein.

[0086] In another preferred embodiment, the molar ratio of aldehyde (A), amine (B), carboxylic acid (C) and isonitrile (D) is 1-2:1-2:1-2:1-2. Preferably, the molar ratio is 1:1:1:1.

[0087] In another preferred embodiment, the reaction is carried out in an organic solvent such as alcohols (e.g., methanol, ethanol), alkanes (e.g., n-hexane, cyclohexane), halogenated hydrocarbons (e.g., dichloromethane, chloroform), aromatic hydrocarbons (e.g., benzene, toluene), nitriles (e.g., acetonitrile), and esters (e.g., ethyl acetate).

[0088] In another preferred embodiment, aldehyde (A) and amine (B) are first stirred at 15-30°C for 10 minutes to 4 hours, and then carboxylic acid (C) and isonitrile (D) are added, and the temperature is raised to 35-60°C and stirred for 4-24 hours.

[0089] In another preferred embodiment, aldehyde (A) can be synthesized by the following method:

[0090] In another preferred embodiment, isonitrile (D) can be synthesized by the following method:

[0091] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0092] The present invention provides a novel cationic lipid compound, its preparation method, composition, and application. The lipid nanoparticles of the present invention have a stable nanostructure, a narrow size distribution, and can be stored at low temperatures for extended periods. They also exhibit good biocompatibility and high in vivo mRNA transfection efficiency. The core structure of the cationic lipid can be constructed via the Ugi reaction, which is simple to operate, operates under mild reaction conditions, does not require additional catalysts, has high atom economy, is simple to implement, and uses inexpensive and readily available raw materials. This not only enhances safety but also facilitates industrial production and quality control. The novel cationic lipid has promising application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] FIG1 shows the cellular delivery efficiency of LNPs prepared from cationic lipids of the present invention. DETAILED DESCRIPTION

[0094] The present invention will be further described below with reference to the following examples, but the present invention is not limited to the following examples. In the examples, if no specific conditions are specified, the reaction was carried out under conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, if the manufacturer is not specified, are conventional products that can be purchased commercially. Unless otherwise specified, percentages and parts are calculated by weight.

[0095] Example 1: Synthesis and characterization of compound 1

[0096] 1) Synthesis of Compound 1A

[0097] Octadecanol (1A-1, 1.0 g, 3.69 mmol) was dissolved in 30 mL of dichloromethane, and PCC (1.2 g, 5.55 mmol) was added. The mixture was allowed to react at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 1A (0.60 g, 95.0%).

[0098] 2) Synthesis of Compound 1D

[0099] Octadecylamine (1D-1, 1.0 g, 3.71 mmol) was dissolved in 10 mL of chloroform, and ethyl formate (2.76 g, 37.1 mmol) was added. The mixture was reacted at 60°C for 16 hours. After the reaction, the excess ethyl formate was removed by concentration, and the product was purified by silica gel column chromatography using a 10 / 1 to 0 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 1D-2 (0.78 g, 70.0%).

[0100] Compound 1D-2 (0.78 g, 2.62 mmol) was dissolved in 25 mL of chloroform, and pyridine (0.63 g, 7.87 mmol) was added. p-Toluenesulfonyl chloride (0.68 g, 3.93 mmol) was added at 0°C, and the reaction was continued with stirring at 0°C for 1 hour. After the reaction, a 5% aqueous sodium sulfate solution was added, and the mixture was stirred for 30 minutes. The mixture was extracted with dichloromethane and concentrated. The crude product was purified by silica gel column chromatography using a 0 / 1 to 20 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to afford product 1D (0.42 g, 57.3%).

[0101] 3) Synthesis of Compound 1

[0102] Compound 1A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 1D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was completed, the reaction solvent was removed by rotary evaporation, and the final product 1 was purified by silica gel column chromatography using dichloromethane:methanol 50 / 1-10:1 (v / v) as the eluent to obtain the final product 1. LCMS: [M+1] + =839.6; 1 H NMR (400 MHz, CDCl3) δ = 6.64 (br s, 1H), 4.76-4.65 (m, 1H), 3.64-3.52 (m, 1H), 3.43-3.32 (m, 2H), 3.09 (s, 3H), 2.52-2.33 (m, 4H), 2.29-2.18 (m, 7H), 2.08-1.85 (m, 4H), 1.66-1.59 (m, 2H), 1.55-1.40 (m, 5H), 1.26 (br s, 62H), 0.89 (br t, J = 6.8 Hz, 6H), pale yellow solid.

[0103] Example 2: Synthesis and characterization of compound 2

[0104] 1) Synthesis of compound 2A

[0105] Compound 2A-1 (1.0 g, 3.72 mmol) was dissolved in 30 mL of dichloromethane, and PCC (1.2 g, 5.59 mmol) was added. The mixture was allowed to react at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 2A (0.970 g, 98.0%).

[0106] 2) Synthesis of compound 2D

[0107] Compound oleylamine (2D-1, 1.0 g, 3.74 mmol) was dissolved in 10 mL of chloroform, and ethyl formate (2.77 g, 37.4 mmol) was added. The mixture was reacted at 60°C for 16 hours. After the reaction, the excess ethyl formate was removed by concentration, and the product was purified by silica gel column chromatography using a 10 / 1 to 0 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 2D-2 (0.78 g, 71.0%).

[0108] Compound 2D-2 (0.78 g, 2.66 mmol) was dissolved in 25 mL of chloroform, and pyridine (0.63 g, 7.97 mmol) was added. p-Toluenesulfonyl chloride (0.76 g, 3.98 mmol) was added at 0°C, and the reaction was continued with stirring at 0°C for 1 hour. After the reaction, a 5% aqueous sodium sulfate solution was added, and the mixture was stirred for 30 minutes. The mixture was extracted with dichloromethane and concentrated. The crude product was purified by column chromatography on silica gel with a 50 / 1 to 20 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain product 2D (0.44 g, 60.0%).

[0109] 3) Synthesis of Compound 2

[0110] Compound 2A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 2D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the final product 2 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent. LCMS: [M+1] + =834.6; 1 H NMR (500 MHz, CDCl3) δ 6.64 (br s, J = 6.0 Hz, 1H), 5.45-5.24 (m, 4H), 4.71 (br t, J = 8.4 Hz, 1H), 3.67-3.53 (m, 1H), 3.43-3.27 (m, 2H), 3.22-3.07 (m, 4H), 2.97-2.62 (m, 1H), 2.54-2.10 (m, 11H), 2.06-1.86 (m, 9H), 1.79-1.59 (m, 6H), 1.55-1.40 (m, 4H), 1.37-1.14 (m, 45H), 0.88 (t, J = 6.8 Hz, 6H), yellow oil.

[0111] Example 3: Synthesis and characterization of compound 3

[0112] 1) Synthesis of compound 3A

[0113] Compound 3A-1 (1.0 g, 3.30 mmol) and 5-hexen-1-ol (0.36 g, 3.60 mmol) were dissolved in 30 mL of dichloromethane. 4-Dimethylaminopyridine (DMAP, 0.60 g, 4.96 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 0.95 g, 4.96 mmol) were added, and the mixture was stirred at 25°C for 1 hour. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain compound 3A-2 (1.03 g, 81.0%).

[0114] Compound 3A-2 (1.03 g, 2.68 mmol) was dissolved in 20 mL of tetrahydrofuran, and tetrabutylammonium fluoride solution (4 mL, 4.0 mmol) was added. The mixture was reacted at 25°C for 2 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 3A-3 (0.69 g, 95.0%).

[0115] Compound 3A-3 (0.69 g, 2.54 mmol) was dissolved in 20 mL of dichloromethane, and PCC (0.83 g, 3.82 mmol) was added. The mixture was reacted at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 3A (0.53 g, 77.0%).

[0116] 2) Synthesis of compound 3

[0117] Compound 3A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 1D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was monitored on a silica gel chromatography plate, the reaction solvent was removed by rotary evaporation, and the final product 3 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 3. LCMS: [M+1] + =839.5; 1H NMR(500MHz, CDCl3)δ6.62(br s,1H),5.78(ddt,J=16.9,10.2,6.7Hz,1H),5.05-4.91(m,2H),4.68(br s,1H),4.05(t,J=6.7Hz,2H),3.56(ddt,J=9.5,6.2,3.1Hz,1H),3.38-3.23(m,2H),3.21

[0118] -3.06 (m, 4H), 2.49-2.33 (m, 3H), 2.31-2.15 (m, 10H), 2.09-2.05 (m, 2H), 1.97-1.86 (m, 2H), 1.76-1.40 (m, 17H), 1.35-1.13 (m, 40H), 0.87 (t, J = 6.9 Hz, 3H), yellow oil.

[0119] Example 4: Synthesis and characterization of compound 4

[0120] 1) Synthesis of compound 4A

[0121] Compound 4A-1 (1.0 g, 3.30 mmol) and 7-tridecanol (0.72 g, 3.60 mmol) were dissolved in 30 mL of dichloromethane. 4-Dimethylaminopyridine (DMAP, 0.60 g, 4.96 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 0.95 g, 4.96 mmol) were added, and the mixture was stirred at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to afford compound 4A-2 (1.21 g, 76.0%).

[0122] Compound 4A-2 (1.21 g, 2.51 mmol) was dissolved in 20 mL of tetrahydrofuran, and tetrabutylammonium fluoride solution (3.76 mL, 3.76 mmol) was added. The mixture was reacted at 25°C for 2 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 4A-3 (0.86 g, 92.1%).

[0123] Compound 4A-3 (0.86 g, 2.30 mmol) was dissolved in 20 mL of dichloromethane, and PCC (0.75 g, 3.46 mmol) was added. The mixture was reacted at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 4A (0.60 g, 70.0%).

[0124] 2) Synthesis of compound 4

[0125] Compound 4A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 1D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 4. LCMS: [M+1] + =939.6; 1 H NMR (500 MHz, CDCl3) δ 6.62 (br s, 1H), 4.85 (t, J = 6.3 Hz, 1H), 4.69 (br t, J = 7.8 Hz, 1H), 3.60-3.54 (m, 1H), 3.39-3.22 (m, 2H), 3.21-3.08 (m, 4H), 2.49-2.33 (m, 3H), 2.28-2.20 (m, 10H), 2.01-1.84 (m, 2H), 1.81-1.54 (m, 9H), 1.55-1.36 (m, 8H), 1.33-1.13 (m, 56H), 0.88-0.85 (m, 9H), yellow oil.

[0126] Example 5: Synthesis and characterization of compound 5

[0127] 1) Synthesis of compound 5D

[0128] Tert-Butoxycarbonyl-11-aminoundecanoic acid (5D-1, 5.0 g, 16.6 mmol) and 7-tridecanol (3.66 g, 18.2 mmol) were dissolved in 80 mL of dichloromethane. 4-Dimethylaminopyridine (DMAP, 3.04 g, 24.9 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 3.82 g, 19.9 mmol) were added, and the mixture was stirred at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by column chromatography on silica gel using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain the product 5D-2 (7.9 g, 98.4%).

[0129] Compound 2B (7.9 g, 16.3 mmol) was dissolved in 100 mL of dichloromethane, and trifluoroacetic acid (TFA, 77.0 g, 675 mmol) was added at 0°C. The reaction was continued at 0°C for 30 minutes. After the reaction, the solvent was removed by concentration, and the pH was adjusted to approximately 10 with saturated aqueous sodium carbonate solution. The mixture was extracted with dichloromethane, and the organic phases were combined and concentrated to obtain the crude product 5D-3 (5.9 g), which was used directly in the next step.

[0130] Compound 5D-3 (5.9 g, 15.4 mmol) was dissolved in 10 mL of chloroform, and ethyl formate (11.4 g, 154 mmol) was added. The mixture was reacted at 60°C for 16 hours. After the reaction, the excess ethyl formate was removed by concentration, and the mixture was purified by silica gel column chromatography using a 10 / 1 to 0 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain product 5D-4 (5.0 g, 78.9%).

[0131] Compound 5D-4 (2.5 g, 6.07 mmol) was dissolved in 25 mL of chloroform, and pyridine (1.44 g, 18.2 mmol) was added. p-Toluenesulfonyl chloride (1.74 g, 9.11 mmol) was added at 0°C, and the reaction was continued with stirring at 0°C for 1 hour. After the reaction, a 5% aqueous sodium sulfate solution was added, and the mixture was stirred for 30 minutes. The mixture was extracted with dichloromethane and concentrated. The crude product was purified by column chromatography on silica gel with a 50 / 1 to 20 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain product 5D (1.3 g, 54.4%).

[0132] 3) Synthesis of Compound 5

[0133] Compound 2A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, and 5.0 mL of anhydrous methanol was added. The mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added. The temperature was raised to 40°C and the reaction was allowed to proceed for 10 hours. After the reaction was monitored on a silica gel chromatography plate, the reaction solvent was removed by rotary evaporation. The product 5 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 5. LCMS: [M+1] + =991.8; 1 H NMR (500 MHz, CDCl3) δ 6.66 (br s, 1H), 5.45-5.18 (m, 2H), 4.87 (p, J = 6.3 Hz, 1H), 4.69 (br t, J = 7.8 Hz, 1H), 3.70-3.53 (m, 1H), 3.40-3.23 (m, 2H), 3.20-3.08 (m, 4H), 2.50-2.36 (m, 6H), 2.29-2.25 (m, 4H), 2.11-1.85 (m, 6H), 1.81-1.54 (m, 12H), 1.54-1.38 (m, 9H), 1.37-1.17 (m, 53H), 0.89-0.86 (m, 9H), yellow oil.

[0134] Example 6: Synthesis and characterization of compound 6

[0135] 1) Synthesis of Compound 6A

[0136] Compound 6A-1 (1.0 g, 3.75 mmol) was dissolved in 30 mL of dichloromethane, and PCC (1.2 g, 5.63 mmol) was added. The mixture was allowed to react at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by silica gel column chromatography using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 6A (0.94 g, 95.1%).

[0137] 2) Synthesis of Compound 6

[0138] Compound 6A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 h. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added. The temperature was raised to 40°C and the reaction was allowed to proceed for 10 h. After completion of the reaction, the solvent was removed by rotary evaporation and the final product 6 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent. LCMS: [M+1]+ =949.7; 1 H NMR (500 MHz, CDCl3) δ 6.64 (br s, 1H), 5.47-5.27 (m, 4H), 4.94-4.83 (m, 1H), 4.70 (br s, 1H), 3.65-3.54 (m, 1H), 3.44-3.22 (m, 2H), 3.21-3.08 (m, 4H), 2.82-2.66 (m, 2H), 2.54-2.36 (m, 4H), 2.34-2.20 (m, 8H), 2.08-1.87 (m, 6H), 1.78-1.58 (m, 7H), 1.56-1.41 (m, 8H), 1.39-1.19 (m, 45H), 0.99-0.80 (m, 9H), yellow oil.

[0139] Example 7: Synthesis and characterization of compound 7

[0140] 1) Synthesis of compound 7

[0141] Compound 4A (1.0 mmol) and compound 1B (1.0 mmol) were placed in a 10 mL reaction flask, and 5 mL of anhydrous methanol was added. The mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added. The temperature was raised to 40°C and the reaction was allowed to proceed for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation. The mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 7. LCMS [M+1] + =1053.8; 1 H NMR (400 MHz, CDCl3) δ = 6.72-6.56 (m, 1H), 4.87 (quin, J = 6.0 Hz, 2H), 4.79-4.63 (m, 1H), 3.64-3.55 (m, 1H), 3.43-3.28 (m, 2H), 3.27-3.08 (m, 4H), 2.52-2.34 (m, 8H), 2.33-2.25 (m, 6H), 1.99-1.90 (m, 10H), 1.76-1.66 (m, 6H), 1.51 (br d, J = 6.0 Hz, 10H), 1.27 (br d, J = 6.8 Hz, 54H), 0.88 (br t, J = 6.8 Hz, 12H), yellow oil.

[0142] Example 8: Synthesis and characterization of compound 8

[0143] 1) Synthesis of compound 8

[0144] Compound 4A (1.0 mmol) and compound 8B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added and the temperature was raised to 40°C for 10 hours. After the reaction was completed, the solvent was removed by rotary evaporation and purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 8. All other compounds can be synthesized using the same method. LC-MS: [M+1] + =1081.9; 1 H NMR (400 MHz, CDCl3) δ = 6.62 (br s, 1H), 4.87 (t, J = 6.4 Hz, 2H), 4.71 (br t, J = 6.8 Hz, 1H), 3.41-3.23 (m, 2H), 3.21-3.09 (m, 4H), 2.61-2.38 (m, 8H), 2.28 (dt, J = 2.8, 7.6 Hz, 5H), 2.00-1.84 (m, 8H), 1.79-1.64 (m, 8H), 1.51 (br d, J = 6.0 Hz, 10H), 1.35-1.22 (m, 54H), 1.04 (br t, J = 6.8 Hz, 6H), 0.93-0.81 (m, 12H), yellow oil.

[0145] Example 9: Synthesis and characterization of compound 9

[0146] 1) Synthesis of compound 9

[0147] Compound 4A (1.0 mmol) and compound 5B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 9. LCMS: [M+1] + =1094.0; 1H NMR (400 MHz, CDCl3) δ = 6.58 (br s, 1H), 4.86-4.73 (m, 2H), 4.61 (br t, J = 7.2 Hz, 1H), 3.57-3.46 (m, 1H), 3.34-3.15 (m, 2H), 3.13-3.00 (m, 3H), 2.29 (br d, J = 6.4 Hz, 6H), 2.26-2.16 (m, 6H), 1.92-1.78 (m, 3H), 1.68 (br s, 4H), 1.57-1.48 (m, 10H), 1.47-1.33 (m, 16H), 1.28-1.15 (m, 54H), 0.89-0.76 (m, 12H), yellow oil.

[0148] Example 10: Synthesis and characterization of compound 10

[0149] 1) Synthesis of compound 10

[0150] Compound 4A (1.0 mmol) and compound 10B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 10. LCMS: [M+1] + =1108.7; 1 H NMR (400 MHz, CDCl3) δ = 6.57 (br s, 1H), 4.79 (quin, J = 6.4 Hz, 2H), 4.63 (br t, J = 6.4 Hz, 1H), 3.56-3.45 (m, 2H), 3.35-3.17 (m, 3H), 3.16-3.02 (m, 5H), 2.45-2.35 (m, 6H), 2.30-2.24 (m, 8H), 2.23-2.17 (m, 6H), 1.91-1.80 (m, 4H), 1.68-1.61 (m, 6H), 1.43 (br d, J = 6.0 Hz, 12H), 1.27-1.14 (m, 54H), 0.81 (t, J = 6.8 Hz, 12H), yellow oil.

[0151] Example 11: Synthesis and characterization of compound 11

[0152] 1) Synthesis of compound 11

[0153] Compound 4A (1.0 mmol) and compound 11B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After the reaction was monitored on a silica gel chromatography plate, the solvent was removed by rotary evaporation, and the final product 11 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent. LCMS: [M+1] + =1109.3; 1 H NMR (400 MHz, CDCl3) δ = 6.60 (br s, 1H), 4.87 (quin, J = 6.0 Hz, 2H), 4.71 (br s, 1H), 3.63-3.56 (m, 1H), 3.32-3.09 (m, 6H), 2.60 (br s, 2H), 2.56-2.24 (m, 10H), 2.00-1.86 (m, 2H), 1.73-1.57 (m, 14H), 1.56-1.45 (m, 14H), 1.27 (br d, J = 7.3 Hz, 54H), 01.09 (br s, 6H), 0.92-0.84 (m, 12H), yellow oil.

[0154] Example 12: Synthesis and characterization of compound 12

[0155] 1) Synthesis of compound 12

[0156] Compound 4A (1.0 mmol) and compound 12B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 5D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation, and the final product 12 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent. LCMS: [M+1] + =1113.8; 1H NMR (400MHz, CDCl3) δ = 6.66 (br s, 1H), 4.91-4.83 (m, 2H), 4.82-4.73 (m, 1H), 3.83-3.65 (m, 4H), 3.50 (br d, J = 4.0 Hz, 2H), 3.47-3.32 (m, 2H), 3.30-3.09 (m, 5H), 2.96-2.89 (m, 1H), 2.81-2.74 (m, 2H), 2.74-2.61 (m, 4H), 2.53-2.42 (m, 4H), 2.42-2.33 (m, 4H), 2.28 (dt, J = 4.0, 7.6 Hz, 5H), 1.99-1.87 (m, 3H), 1.77-1.70 (m, 5H), 1.51 (br d, J = 5.6 Hz, 10H), 1.27 (br s, 54H), 0.92-0.84 (m, 12H)., yellow oil.

[0157] Example 13: Synthesis and characterization of compound 13

[0158] 1) Synthesis of compound 13

[0159] Compound 2A (1.0 mmol) and compound 8B (1.0 mmol) were placed in a 10 mL reaction flask, 5 mL of anhydrous methanol was added, and the mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 2D (1.0 mmol) were then added, and the temperature was raised to 40°C for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation, and the mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 13. LCMS: [M+1] + =863.8; 1 H NMR (500 MHz, CDCl3) δ 6.63 (br s, 1H), 5.46-5.23 (m, 4H), 4.71 (br s, 1H), 3.69-3.52 (m, 1H), 3.43-3.23 (m, 2H), 3.22-3.08 (m, 4H), 2.92-2.68 (m, 1H), 2.60-2.31 (m, 9H), 2.07-1.85 (m, 9H), 1.79–1.55 (m, 6H), 1.52-1.39 (m, 4H), 1.38-1.16 (m, 45H), 1.02 (t, J = 7.1 Hz, 6H), 0.88 (t, J = 6.8 Hz, 6H), yellow oil.

[0160] Example 14: Synthesis and characterization of compound 14

[0161] 1) Synthesis of compound 14

[0162] Compound 6A (1.0 mmol) and compound 8B (1.0 mmol) were placed in a 10 mL reaction flask, and 5 mL of anhydrous methanol was added. The mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 2D (1.0 mmol) were then added. The temperature was raised to 40°C and the reaction was allowed to proceed for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation. The mixture was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product 15. LCMS: [M+1] + =861.5; 1 H NMR (500 MHz, CDCl3) δ 6.62 (br s, 1H), 5.48-5.21 (m, 6H), 4.72 (br s, 1H), 3.65-3.53 (m, 1H), 3.43-3.23 (m, 2H), 3.23-3.03 (m, 4H), 2.92-2.61 (m, 1H), 2.58-2.31 (m, 9H), 2.16-1.87 (m, 9H), 1.78-1.57 (m, 6H), 1.53-1.39 (m, 4H), 1.38-1.20 (m, 41H), 1.02 (t, J = 7.1 Hz, 6H), 0.88 (t, J = 6.8 Hz, 6H), yellow oil.

[0163] Example 15: Synthesis and characterization of compound 15

[0164] 1) Synthesis of Compound 15A

[0165] N-tert-Butyloxycarbonyl-8-aminooctanoic acid (15A-1, 1.0 g, 3.86 mmol) and cis-2-nonen-1-ol (0.6 g, 4.24 mmol) were dissolved in 20 mL of dichloromethane. 4-Dimethylaminopyridine (DMAP, 0.71 g, 5.78 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 1.11 g, 5.78 mmol) were added, and the mixture was stirred at 25°C for 3 hours. After completion of the reaction, the solvent was removed by concentration, and the product was purified by column chromatography on silica gel using a 50 / 1 to 10 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain product 15A-2 (1.42 g, 96.0%).

[0166] Compound 15A-2 (1.42 g, 16.3 mmol) was dissolved in 100 mL of dichloromethane, and trifluoroacetic acid (TFA, 17.7 g, 155 mmol) was added at 0°C. The reaction was continued at 0°C for 30 minutes. After the reaction, the solvent was removed by concentration, and the pH was adjusted to approximately 10 with saturated aqueous sodium carbonate solution. The mixture was extracted with dichloromethane, and the organic phases were combined and concentrated to obtain the crude product 15A-3 (1.0 g), which was used directly in the next step.

[0167] Compound 15A-3 (1.0 g, 3.70 mmol) was dissolved in 10 mL of chloroform, and ethyl formate (2.74 g, 37.0 mmol) was added. The mixture was reacted at 60°C for 16 hours. After the reaction, the excess ethyl formate was removed by concentration, and the mixture was purified by silica gel column chromatography using a 10 / 1 to 0 / 1 (v / v) ratio of petroleum ether to ethyl acetate to afford product 15A-4 (1.0 g, 86.9%).

[0168] Compound 15A-4 (1.0 g, 3.21 mmol) was dissolved in 25 mL of chloroform, and pyridine (0.76 g, 9.63 mmol) was added. p-Toluenesulfonyl chloride (0.92 g, 4.82 mmol) was added at 0°C, and the reaction was continued with stirring at 0°C for 1 hour. After the reaction, a 5% aqueous sodium sulfate solution was added, and the mixture was stirred for 30 minutes. The mixture was extracted with dichloromethane and concentrated. The crude product was purified by column chromatography on silica gel with a 50 / 1 to 20 / 1 (v / v) ratio of petroleum ether to ethyl acetate as the eluent to obtain product 15A (0.53 g, 52.7%).

[0169] 2) Synthesis of compound 15

[0170] Compound 4A (1.0 mmol) and compound 8B (1.0 mmol) were placed in a 10.0 mL reaction flask, and 5 mL of anhydrous methanol was added. The mixture was stirred at room temperature for 0.5 hours. Compound 1C (1.0 mmol) and compound 15D (1.0 mmol) were then added. The temperature was raised to 40°C and the reaction was allowed to proceed for 10 hours. After completion of the reaction, the solvent was removed by rotary evaporation. The product 15 was purified by silica gel column chromatography using dichloromethane:methanol (20:1, v / v) as the eluent to obtain the final product. LCMS: [M+1] + =980.6; 1HNMR(400MHz, CDCl3)δ=6.64(br s,1H),5.70-5.62(m,1H),5.58-5.51(m,1H),4.88(quin,J=6.4Hz,1H),4. 75-4.67(m,1H),4.64(d,J=6.8Hz,2H),3.64-3.57(m,1H),3.48-2.87(m,6H ),2.64-2.21(m,12H),2.12(q,J=7.2Hz,2H),2.02-1.86(m,2H),1.81-1.66 (m,6H),1.65-1.57(m,8H),1.56-1.46(m,8H),1.39-1.24(m,40H),1.04(br d, J = 6.5 Hz, 4H), 0.93-0.87 (m, 9H), yellow oil.

[0171] Example 16: In vitro evaluation of prepared luciferase mRNA lipid nanoparticles (LNPs)

[0172] HeLa cells in the logarithmic growth phase were seeded at 60,000 cells per well in a 48-well cell culture plate and cultured overnight until the cells adhered. A novel cationic lipid, DOPE (purchased from Shanghai Avto Pharmaceutical Technology Co., Ltd.), cholesterol (purchased from Shanghai Avto Pharmaceutical Technology Co., Ltd.), and DMG-PEG2000 (purchased from Xiamen Sinobond Biotechnology Co., Ltd.) were dissolved in ethanol at a molar ratio of 50:10:38.5:1.5. Luciferase mRNA (purchased from Hefei Afana Biotechnology Co., Ltd.) was dissolved in a certain volume of RNase-free citrate buffer (pH = 3-5) at a dose of 250 ng per well. The ethanol phase (organic phase) and the buffer phase (aqueous phase) were quickly mixed at a volume ratio of 1 / 3 and incubated for 15 minutes to obtain LNPs encapsulating firefly luciferase mRNA. At the same time, the culture medium was replaced with fresh PRMI1640 culture medium. After the incubation, the prepared LNPs were diluted with 1xPBS to an ethanol concentration of less than 5% and added to the cell culture plate. After 6 hours, the culture medium was replaced with DMEM culture medium and cultured for 24 hours. The cells were treated with a firefly luciferase reporter gene detection kit (purchased from Shanghai Biyuntian Biotechnology Co., Ltd.). The fluorescence intensity was measured by 20 / 20 Luminometer. Each experiment was repeated three times and the data were averaged. The results are shown in Table 2 and Figure 1.

[0173] Table 2 Fluorescence intensity results after mRNA-LNP transfection into Hela cells

[0174] In vitro cell transfection experiments demonstrated that LNPs prepared using the cationic lipids of the present invention exhibited good biocompatibility and high in vitro mRNA transfection efficiency, with LNPs prepared using cationic lipid 7 achieving the highest delivery efficiency. Therefore, LNPs can be used to deliver different therapeutic agents, tailored to the needs of diverse therapeutic environments.

[0175] Example 17: In vivo evaluation of prepared luciferase mRNA lipid nanoparticles (LNPs):

[0176] A novel cationic lipid, DOPE (purchased from Shanghai Avto Pharmaceutical Technology Co., Ltd.), cholesterol (purchased from Shanghai Avto Pharmaceutical Technology Co., Ltd.), and DMG-PEG2000 (purchased from Xiamen Sinobond Biotechnology Co., Ltd.) were dissolved in anhydrous ethanol at a molar ratio of 50:10:38.5:1.5. A certain amount of firefly luciferase mRNA (purchased from Hefei Afana Biotechnology Co., Ltd.) was dissolved in a certain volume of RNase-free citrate buffer (pH = 3-5), ensuring a 1 / 3 volume ratio of ethanol phase (organic phase) to buffer phase (aqueous phase). The two phases were rapidly mixed using a pipette to produce LNPs encapsulating firefly luciferase mRNA. The mixture was incubated at room temperature for 15 minutes, then dialyzed against PBS buffer for two hours using dialysis tubing. The mRNA-LNPs were collected and studied in 6-8 week old female Balb / c mice. The lipid nanoparticles were systemically administered with a dose of 0.25 mg / kg luciferase mRNA via tail vein injection. At a specific time point (3 hours) after administration, 150 μL (10 mg / mL) of luciferin substrate was injected intraperitoneally into the mice. Five minutes later, the mice were placed in a small animal in vivo imaging device to measure the fluorescence intensity. The final results were expressed as the mean fluorescence intensity. Each group of experiments was repeated three times, and the data were averaged. The results are shown in Table 3.

[0177] Table 3 Fluorescence intensity results after tail vein administration in mice

[0178] In vivo transfection experiments demonstrated that LNPs prepared using the cationic lipids of the present invention exhibited good biocompatibility and high mRNA transfection efficiency in vivo, with LNPs prepared using cationic lipids 5 and 8 achieving the highest delivery efficiency. Therefore, they can be used to deliver different therapeutic agents as needed to meet the needs of diverse therapeutic environments.

[0179] At the same time, through comparison of in vitro and in vivo experimental results, it was found that the LNP prepared from the same cationic lipid molecule of the present invention exhibited different transfection efficiencies in vivo and in vitro, and the in vitro nucleic acid delivery level could not accurately reflect the in vivo nucleic acid delivery level, that is, there was no correlation between the in vivo and in vitro nucleic acid delivery efficiencies. This result is consistent with previous relevant research results in this field.

[0180] Example 18: Determination of particle size and polydispersity index (PDI) of prepared nanolipid particles:

[0181] Dynamic light scattering was used to determine the particle size and polydispersity index (PDI) using a Malvern laser particle size analyzer. 10 μL of the LNP solution prepared using the method of Example 17 was taken, diluted to 1 mL with RNase-free deionized water, added to the sample pool, and each sample was measured 3 times. The measurement conditions were: 90° scattering angle, 25°C. The test results are shown in Table 4. The particle size of the prepared LNP was between 100 and 200 nm, and the PDI was between 0 and 0.3, indicating that the particle size of the prepared LNP met the range requirements of lipid nanoparticles in this field, and the particle size distribution was relatively uniform. The particle size can be adjusted according to the needs of the use scenario by using different preparation methods or / and adjusting the proportions of the components of the composition.

[0182] Table 4 Particle size and polydispersity index (PDI) of nanolipid particles

[0183] Based on the results of in vitro and in vivo experiments and physicochemical property measurements, the LNPs prepared from the cationic lipids of the present invention have achieved good biocompatibility and high in vivo mRNA transfection efficiency. The cationic lipids of the present invention can form stable nanostructures with a narrow size distribution. The size varies with the structure of different liposomes and is within the range of 200 to 300 nm. Moreover, the LNPs formed by the cationic lipids of the present invention can be stored at low temperatures for a long time. Structurally, the core structure containing amide bonds is introduced through the classic Ugi four-component reaction, which is conducive to the formation of hydrogen bonds between cationic lipid molecules in LNPs and improves the colloidal stability of LNPs; the introduction of degradable ester bonds in the hydrophobic tails of cationic lipid molecules can change the metabolic behavior of liposomes in the body, thereby improving the biosafety of mRNA-LNPs; the introduction of lipoic acid structures on the core nucleus can enhance the cell's ability to uptake LNPs, thereby increasing transfection efficiency. In terms of synthesis, the core structure of the cationic lipids of the present invention can be constructed through the Ugi reaction. This reaction is simple to operate, has mild reaction conditions, does not require additional catalysts, has high atom economy, is simple and easy to perform, and uses cheap and readily available raw materials. It not only has higher safety but also facilitates its industrial production and quality control. The new cationic lipids have good application prospects.

[0184] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0185] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the disclosed patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the scope of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A compound as shown in formula I, or its isomer, pharmaceutically acceptable salt, prodrug; in: Side chain part -W1-L1-(R1) n1 、-W2-L2-(R2) n2 and-W3-L3-(R3) n3 The main chain (i.e., the longest chain) of each independently has 1-40 carbons; W1, W2 and W3 are each independently C1-C14 alkylene, C2-C14 alkenylene or C2-C14 alkynylene; n1, n2, and n3 are each independently 0, 1, or 2; L1, L2 and L3 are each independently selected from the following structures: R1, R2 and R3 are each independently H, aryl, cholesteryl, adamantyl, C6-24 alkyl, C6-24 alkenyl, C6-24 alkynyl, C4-C10 cycloalkyl, C4-C10 cycloalkenyl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl are arbitrarily connected by one or more ester bonds, amide bonds, carbonate bonds, amino acid ester bonds or ether bonds; R1, R2 and R3 may also be selected as a 4-10 membered heterocyclic ring, wherein the heterocyclic ring includes one or more heteroatoms selected from N, O, S and Se; R4 and R5 are each independently C1-C12 alkyl, C2-C12 alkenyl or C2-C12 alkynyl, and the alkyl, alkenyl or alkynyl is arbitrarily substituted by one or more OH, SH, C1-C4 alkoxy, or R4 and R5 are combined with each other to form a 4-10 membered heterocyclic ring, and in addition to the N atom connected to M, the heterocyclic ring optionally includes one or more heteroatoms of N, O, and S; M is a C1-C12 alkylene group, a C2-C12 alkenylene group or a C2-C12 alkynylene group.

2. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, prodrug, characterized in that: The compound of formula I has a structure shown in the following formula II: X, Y and Z are each independently CH2, NH, S, Se; represents a single or double bond; m and n are each independently 0, 1, 2, 3 or 4; The definitions of the remaining groups are as described in claim 1; Preferably, the compound has the structure shown in Formula I and the compound has the structure shown in Formula III below: The definitions of the groups are as described in claim 1.

3. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, prodrug, characterized in that: Side chain part -W1-L1-(R1) n1 、-W2-L2-(R2) n2 and-W3-L3-(R3) n3 The main chain (i.e., the longest chain) of each independently has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 carbons; W1, W2 and W3 are each independently C3-C8 alkyl, C3-C8 alkenyl or C3-C8 alkynyl; preferably, W1, W2 and W3 are each independently C3-C8 alkyl, C3-C8 alkenyl or C3-C8 alkynyl; R1, R2 and R3 are each independently H, C3-C22 alkyl, C3-C22 alkenyl, C3-C22 alkynyl, C5-C8 cycloalkyl, C5-C8 cycloalkenyl; preferably, R1, R2 and R3 are each independently H, C6-C14 alkyl, C6-C14 alkenyl, C6-C14 alkynyl; Most preferably, the side chain moieties -W1-L1-R1 and -W2-L2-R2 are each independently selected from the following structures:

4. The compound according to any one of claims 1 to 2 or its isomer, pharmaceutically acceptable salt, prodrug, characterized in that: R4 and R5 are each independently C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, and the alkyl, alkenyl or alkynyl is arbitrarily substituted by one or more OH, C1-C4 alkoxy, or R4 and R5 are combined with each other to form a 5-8 membered heterocyclic ring, and in addition to the N atom connected to M, the heterocyclic ring optionally includes one or more heteroatoms of N, O, and S; preferably, R4 and R5 are each independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 2-hydroxyethyl, 3-hydroxypropyl, or R4 and R5 are combined with each other to form one of the following structures: Wherein, R is selected from one of C(Ra)2, NRb, O, and S; Ra are each independently H, OH, C1-12 alkyl; Rb are each independently H, C1-12 alkyl, C(O)C1-12 alkyl; a1 is 1, 2, 3, 4, 5, 6, 7 or 8; a2 is 1, 2, 3, 4, 5 or 6; a3 is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; preferably, R4 and R5 are combined with each other to form tetrahydropyrrol-1-yl, piperidinyl-1-yl, piperazin-1-yl, 4-methylpiperazin-1-yl; M is C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene; preferably, M is -(CH2) s -, s is 1, 2, 3, 4, 5 or 6; Preferably, the -MN(R4)(R5) has one of the following structures:

5. The compound according to any one of claims 1 to 2 or its isomer, pharmaceutically acceptable salt, prodrug, characterized in that: The compound is selected from the compounds shown in Table 1.

6. A lipid nanoparticle composition comprising a compound according to any one of claims 1 to 5 or one or more of its isomers, pharmaceutically acceptable salts, prodrugs and a therapeutic and / or preventive agent.

7. The lipid nanoparticle composition according to claim 6, characterized in that The lipids also include one or more of neutral lipids, steroids and polymer-conjugated lipids; preferably, the lipid nanoparticle composition is mainly composed of cationic lipids, neutral lipids, steroids and polymer-conjugated lipids in combination with therapeutic agents and / or preventive agents; more preferably, the molar ratio of the compound or its isomer, pharmaceutically acceptable salt, prodrug according to any one of claims 1 to 5, and neutral lipids, steroids and polymer-conjugated lipids in the lipid nanoparticle composition is (25-75): (5-25): (15-65): (0.5-10), preferably (40-60): (5-15): (35-45): (1-2); Among them, the neutral lipid is preferably selected from one or more of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and sphingomyelin (SM); the steroid is preferably selected from one or more of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, and α-tocopherol; the polymer-conjugated lipid is preferably selected from pegylated lipids.

8. The lipid nanoparticle composition according to claim 6, characterized in that The therapeutic and / or preventive agent comprises a nucleic acid; preferably, the nucleic acid is RNA selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA, tRNA, mRNA, and sgRNA; further preferably, the nucleic acid is used to treat a disease by targeting a specific target; optionally, the nucleic acid molecule can be used to treat cancer, inflammation, fibrotic diseases, autoimmune diseases, infections, congenital and hereditary diseases, connective tissue diseases, digestive system diseases, endocrine diseases, eye diseases, reproductive diseases, cardiovascular diseases, kidney and urinary diseases, respiratory system diseases, metabolic disorders, musculoskeletal diseases, nervous system diseases, and blood system diseases.

9. Use of a compound according to any one of claims 1 to 5 or an isomer, a pharmaceutically acceptable salt, a prodrug thereof, or a lipid nanoparticle composition according to any one of claims 6 to 8 in the preparation of a delivery system or in the preparation of a medicament for treating a disease or condition in a patient.

10. A method for delivering a therapeutic agent and / or a preventive agent to a cell in vitro or in vivo, the method comprising administering to the cell a compound according to any one of claims 1 to 5 or an isomer, a pharmaceutically acceptable salt, a prodrug thereof, or a lipid nanoparticle composition according to any one of claims 6 to 8.

11. A method for synthesizing the compound of formula I according to any one of claims 1 to 5, comprising: Reacting an aldehyde (A), an amine (B), a carboxylic acid (C) and an isonitrile (D) to produce a compound of formula I; Wherein, the definitions of R1-R3, W1-W3, L1-L3, M, and n1-n3 are as described in any one of claims 1-5; Preferably, the molar ratio of aldehyde (A), amine (B), carboxylic acid (C) and isonitrile (D) is 1-2:1-2:1-2:1-2.

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