An ionizable cationic lipid compound, and methods of making and using the same

Abstract

This invention provides an ionizable cationic lipid compound, its preparation method, and its application, belonging to the field of biomedical technology. The ionizable cationic lipid compound provided by this invention is a compound having formula (1), or its salt, stereoisomer, or tautomer, and can be used as a surfactant in various fields, especially as a lipid composition for delivering therapeutic agents. The lipid nanoparticles prepared from it can exhibit good efficacy and safety in cells and animals.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an ionizable cationic lipid compound, its preparation method, and its application. Background Technology

[0002] Gene therapy uses nucleic acids as functional molecules to activate biological therapies. Nucleic acids have gained significant attention due to global efforts in elucidating the human genome and recent discoveries such as RNA interference (RNAi) and CRISPR-based genome editing. Gene therapy uses genetic material to alter the expression of target genes or modify the biological characteristics of living cells to meet therapeutic needs. In recent years, several gene therapy products have been approved by regulatory agencies for various applications, such as the use of mRNA vaccines to combat the COVID-19 pandemic. Gene therapy can be divided into three main approaches: 1) editing variant genes using CRISPR-Cas technology; 2) upregulating gene expression by inserting functional gene copies, using molecules including DNA plasmids (pDNA), small circular DNA (mcDNA), synthetic mRNA, circular RNA, and self-amplifying RNA (saRNA); and 3) downregulating gene expression using molecules such as small interfering RNA (siRNA), antisense oligonucleotides (ASO), short hairpin RNA (shRNA), and microRNA (miRNA).

[0003] Nucleic acids, due to their negative charge, high molecular weight, and hydrophilicity, have poor permeability across cell membranes. Furthermore, nucleic acids exhibit low stability in vivo and are rapidly cleared from the host outside the cell. Therefore, delivering nucleic acids to their active sites within cells is one of the most challenging aspects of gene therapy. This is especially true when delivering RNA molecules, as RNA molecules typically have short-term activity and low intracellular retention, requiring more frequent administration. Thus, payload and vector toxicity are two crucial metrics. The vector itself needs to overcome extracellular and intracellular barriers, protect against nuclease activity in the bloodstream, enhance and assist cellular uptake, and facilitate endosome escape after entering the cell.

[0004] Commonly used nucleic acid vectors are divided into two categories: viral vectors and non-viral vectors. Viral gene delivery systems have high cell transfection efficiency, but because the human body produces neutralizing antibodies against the vector or pre-existing antibodies against the viral vector, the vector is rapidly cleared. At the same time, viral vectors are typically small in size (usually less than 7kb). Therefore, lipid-based non-viral gene delivery systems have become the mainstream vector delivery systems.

[0005] Ionizable cationic lipids are an important component of lipid-based nonviral carriers. From a chemical structure perspective, ionizable cationic lipids are amphiphilic molecules, consisting of three parts: a head group, a linker, and a tail chain. The head group typically contains one or more ionizable sites, allowing the molecule to be neutral or positively charged under different pH conditions. Below their pKa, ionizable lipids can form lipophilic drug complexes with oppositely charged nucleic acids, anionic small molecules, peptides, proteins, and heparin. Lipids and nucleic acids can also form lipid nanoparticles (LNPs) through electrostatic interactions.

[0006] In pharmaceutical products, ionizable cationic lipids possess pH-dependent cationic properties, posing certain safety concerns. First-generation cationic lipids (e.g., DOTMA, DOTAP) bind to cell and nuclear membranes, triggering the release of degrading enzymes from lysosomes, leading to mitochondrial permeability and dysfunction, generating reactive oxygen species, altering the function of cytoplasmic enzymes, and damaging DNA. To address this deficiency of cationic lipids, around 2001, Acuitas designed second-generation lipid structures with dimethyl tertiary amine as the head group. Compared to first-generation lipid structures, the electrical properties of lipid structures with dimethyl tertiary amine as the head group can be adjusted by pH. Therefore, starting with the second-generation lipid structures, cationic lipids used as nucleic acid carriers are also referred to as ionizable cationic lipids (see CN105873902B, CN107207428A, CN110003066B, CN110325511B, CN110337429B). Common second-generation ionizable cationic lipids include D-Lin-MC3-DMA, DLin-K-DMA, and DlinDMA (CN107922364B, CN108368028B). Second-generation ionizable lipids have been successfully used as carriers for siRNA drugs. For example, Acuitas Therapeutics successfully developed the lipid D-Lin-MC3-DMA, and its siRNA product Onparrto, developed in collaboration with Alnylam, became the world's first nucleic acid drug, approved for marketing in 2018. Subsequently, Moderna developed third-generation lipids with hydroxyl tertiary amines as head groups (CN110520409A, CN114746398A). Third-generation ionizable lipids exhibit a positive charge under acidic pH conditions, which facilitates complexation with nucleic acids and allows them to escape from endosomes after entering cells. Currently, high-performance ionizable cationic lipids include ALC-0315 (4-hydroxybutyl)azo-2-(hexane-6,1-diyl)bis(2-hexyldecanoate) and lipid H (SM-102) (9-heptadecyl-8-{(2-hydroxyethyl)[6-oxo-6-(undecapoxy)hexyl]amino}octanoate), which are used in the coronavirus (COVID-19) mRNA vaccines BNT162b and mRNA-1273, respectively.

[0007] Most patents currently protect ionizable cationic lipids that introduce head groups and tail chains using a tertiary amine group as the central functional group. Of the three functional groups on the tertiary amine, one is used to introduce the head group, and the other two are used to introduce the tail chain. The ionizable cationic lipid compounds (including their stereoisomers, tautomers, and corresponding salts of these lipids) provided by this invention use a tetrafunctional compound as the core unit, and introduce the corresponding head group and tail chain through the terminal functional group of the tetrafunctional compound. In this invention, one of these four functional groups is used to introduce the head group, and the other three functional groups are used to introduce the tail chain, as shown in formula (1):

[0008]

[0009] The ionizable cationic lipid compounds provided by this invention use ionizable cationic compounds and their analogues as ionizable lipid head groups, lipids such as fatty acids as tail chains, and tetrafunctional compounds as linkers. The ionizable lipid head groups, tail chains, and linkers are connected by biodegradable chemical bonds (such as ester or amide bonds), forming amphiphilic lipid compounds. In constructing the structure of these ionizable cationic lipid compounds, this invention introduces biodegradable functional groups. Because these biodegradable chemical bonds can be cleaved in vivo by various enzymes such as lipases, esterases, and proteases, rapidly hydrolyzing into endogenous metabolites, this reduces cytotoxicity and improves the safety of their use. Summary of the Invention

[0010] This invention addresses the problems existing in the prior art by providing an ionizable cationic lipid compound, its preparation method, and its applications. The ionizable cationic lipid compound provided by this invention uses an ionizable cationic compound or its analogue as the ionizable lipid head group, a lipid such as a fatty acid as the tail chain, and a tetrafunctional compound as a linker. The ionizable lipid head group, tail chain, and linker are connected by biodegradable chemical bonds (such as ester or amide bonds), forming an amphiphilic lipid compound. These biodegradable chemical bonds can be cleaved in vivo by various enzymes such as lipases, esterases, and proteases, rapidly hydrolyzing into endogenous metabolites, exhibiting low cytotoxicity.

[0011] To achieve the above objectives, in a first aspect, the present invention provides an ionizable cationic lipid compound, said ionizable cationic lipid compound being a compound of formula (1), or a salt, stereoisomer, or tautomer thereof:

[0012]

[0013] Among them, R1, R2, and R3 are H and C independently of each other. 5-40 Straight-chain or branched alkyl, C 5-40 Straight-chain or branched alkenyl, C 5-40The side chains are: straight-chain or branched alkynyl groups, 3-6 membered saturated or partially unsaturated cyclic hydrocarbon groups containing 1-3 side chains, or 6-10 membered aromatic groups containing 1-3 side chains; wherein the side chains are independently selected from C... 10-30 Straight-chain or branched alkyl groups, C 10-30 Straight-chain or branched alkenyl, C 10-30 Straight-chain or branched alkynyl group; condition that at most one of R1, R2, and R3 is H;

[0014] M is selected from -NR4R5, saturated or partially unsaturated 3-6 membered heterocyclic groups containing at least one nitrogen atom, and 6-10 membered heteroaryl groups containing at least one nitrogen atom, wherein the heterocyclic group or heteroaryl group is not substituted or is substituted by one or more -OH, carboxyl, amino, oxo, or halogen groups.

[0015] R4 and R5 are H and C, respectively, which are independent of each other. 1-6 Straight-chain or branched alkyl groups, C 2-6 Straight-chain or branched alkenyl or C 2-6 Straight-chain or branched alkynyl groups, wherein the C 1-6 Straight-chain or branched alkyl groups, C 2-6 Straight-chain or branched alkenyl or C 2-6 The straight-chain or branched alkynyl group is either unsubstituted or substituted by one or more -OH, carboxyl, aminoamide, amidine, guanidine, or halogen groups;

[0016] G1, G2, and G3 are independently -O-, -S-, -NR6-, -SS-, -C(=O)-, -C(=S)-, -C(=O)O-, -CH(OH)-, -OC(=O)-, -C(=O)NR6-, -NR6C(=O)-, -OC(=O)O-, -NR6C(=O)O-, -OC(=O)NR6-, -NR6C(=O)NR 13 -, -C(=O)S-, -C(=S)S-, -SC(=S)-, -SC(=O)-, -OC(=O)S-, -SC(=O)O-, -SC(=O)S-, -OS(=O)2O-, -S(=O)2O-, -OS (=O)2-, -S(=O)2-, -S(=O)2-NR6-, -NR6-S(=O)2-, -P(=O)(OR6)O-, -OP(=O)(OR6)- or -OP(=O)(OR6)O-; where each R6, R 13 They are independently selected from H, hydroxyl, and C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-30 Alkenes, whether straight-chain or branched;

[0017] L1 is selected from -X1- or -(CR7R8) m-X1-, where each X1 is independently selected from -O-, -S-, -NR 14 -, -SS-, -C(=O)-, -C(=S)-, -C(=O)O-, -OC(=O)-, -C(=O)NR 14 -、-NR 14 C(=O)-、-OC(=O)O-、-NR 14 C(=O)O-、-OC(=O)NR 14 -、-NR 14 C(=O)NR 15 -, -C(=O)S-, -C(=S)S-, -SC(=S)-, -SC(=O)-, -OC(=O)S-, -SC(=O)O-, -SC(=O)S-, -OS(=O)2O-, -S(=O)2O-, -OS(=O)2-, -S(=O)2-, -S(=O)2-NR 14 -、-NR 14 -S(=O)2-、-P(=O)(OR 14 )O-、-OP(=O)(OR 14 - or -OP(=O)(OR 14 )O-; where m is an integer from 2 to 6, and R7 and R8 are independently H, hydroxyl, halogen, C 1-6 Straight-chain or branched alkyl or cycloalkyl, C 2-6 Straight-chain or branched alkenyl groups, each R 14 R 15 They are selected independently from H and C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-30 Alkenes, whether straight-chain or branched;

[0018] L2 is -(CR9R) 10 ) n -or-(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - where X2 is selected from -O-, -S-, -NR 16 -, -SS-, -C(=O)-, -C(=S)-, -C(=O)O-, -OC(=O)-, -C(=O)NR 16 -、-NR 16 C(=O)-、-OC(=O)O-、-NR 16 C(=O)O-、-OC(=O)NR 16 -、-NR 16 C(=O)NR 17-, -C(=O)S-, -C(=S)S-, -SC(=S)-, -SC(=O)-, -OC(=O)S-, -SC(=O)O-, -SC(=O)S-, -OS(=O)2O-, -S(=O)2O-, -OS(=O)2-, -S(=O)2-, -S(=O)2-NR 16 -、NR 16 -S(=O)2-、-P(=O)(OR 16 )O-、-OP(=O)(OR 16 - or -OP(=O)(OR 16 O-; n is an integer from 1 to 6; k is an integer from 1 to 6; R9, R 10 R 11 R 12 The elements that are independent of each other are H, hydroxyl, halogen, and C. 1-6 Straight-chain or branched alkyl or cycloalkyl, C 2-6 Straight-chain or branched alkenyl groups, each R 16 R 17 They are selected independently from H and C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-30 Alkenes, whether straight-chain or branched;

[0019] Among them, R4 to R 17 The alkyl, cycloalkyl, and alkenyl groups mentioned herein are either not substituted or are substituted by one or more groups selected from hydroxyl, mercapto, amino, substituted amino, and halogen groups;

[0020] The salts mentioned do not include quaternary ammonium salts.

[0021] In a preferred embodiment, R1, R2, and R3 are independently groups of the following:

[0022]

[0023] Among them, Y does not exist, or it is C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-20 Straight-chain or branched alkenyl, C 2-20 Straight-chain or branched alkynyl group; R1' and R2' are independently H and C. 1-30 Straight-chain or branched alkyl groups, C 2-30 Straight-chain or branched alkenyl, C 2-30 Straight-chain or branched alkynyl groups, with a total carbon chain length of 8-40 for Y, R1' and R2'.

[0024] In a preferred embodiment, R1, R2, and R3 are independently selected from the following groups:

[0025]

[0026] Among them, R1' and R2' are H and C, respectively, which are independent of each other. 1-30 Straight-chain or branched alkyl groups, C 2-30 Straight-chain or branched alkenyl, C 2-30 Straight-chain or branched alkynyl groups, with a total carbon chain length of 8-30 for R1' and R2'.

[0027] In a preferred embodiment, R1, R2, and R3 are independently selected from any one of the following groups:

[0028]

[0029] In a preferred embodiment, G1, G2, and G3 are independently -O-, -S-, -NR6-, -SS-, -C(=O)-, -C(=O)O-, -CH(OH)-, -OC(=O)-, -C(=O)NR6-, -NR6C(=O)-, -OC(=O)O-, -NR6C(=O)O-, -OC(=O)NR6-, -NR6C(=O)NR 13 -, -P(=O)(OR6)O-, -OP(=O)(OR6)- or -OP(=O)(OR6)O-.

[0030] In a preferred embodiment, L1 is selected from -(CR7R8). m -X1-, where X1 is selected from -O-, -S-, -NR 14 -, -SS-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR 14 -、-NR 14 C(=O)-、-OC(=O)O-、-NR 14 C(=O)O-、-OC(=O)NR 14 -、-NR 14 C(=O)NR 15 -、-P(=O)(OR 14 )O-、-OP(=O)(OR 14 - or -OP(=O)(OR 14 )O-.

[0031] In a preferred embodiment, L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - where X2 is selected from -O-, -S-, -NR 16-, -SS-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR 16 -、-NR 16 C(=O)-、-OC(=O)O-、-NR 16 C(=O)O-、-OC(=O)NR 16 -、-NR 16 C(=O)NR 17 -、-P(=O)(OR 16 )O-、-OP(=O)(OR 16 - or -OP(=O)(OR 16 )O-.

[0032] In a preferred embodiment, M is selected from the following structures:

[0033]

[0034] Where m' and n' are independent integers from 0 to 6, and R1” and R2” are independent integers H and C. 1-6 alkyl, C 2-6 The nitrogen-containing heterocycle is selected from pyrrole, imidazole, pyridine, pyrazole, triazole, oxazole, isoxazole, thiophene, isothiazole, pyridazine, pyrazine, piperazine, indole, benzimidazole, carbazole, quinoline, isoquinoline, purine, and pyrimidine, and their tautomeric forms, and is not substituted or optionally is substituted by one or more groups selected from hydroxyl, mercapto, amino, substituted amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 6-14 The aryl group is substituted with an organic group.

[0035] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1A):

[0036]

[0037] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1B):

[0038]

[0039] In a preferred embodiment, the compound of formula (1) is selected from the compounds of formula (1C):

[0040]

[0041] In a preferred embodiment, the compound of formula (1) is selected from compounds of formula (1D):

[0042]

[0043] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1E).

[0044]

[0045] In a preferred embodiment, the compound of formula (1) is selected from the compounds represented by formula (1F):

[0046]

[0047] In a preferred embodiment, the compound of formula (1) is selected from the compounds represented by formula (1G):

[0048]

[0049] In a preferred embodiment, the compound of formula (1) is selected from the compounds represented by formula (1H):

[0050]

[0051] In a preferred embodiment, M is selected from any one of the following groups:

[0052]

[0053] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1I):

[0054]

[0055] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1J):

[0056]

[0057] In a preferred embodiment, the compound of formula (1) is selected from the compounds shown in formula (1K):

[0058]

[0059] In a preferred embodiment, Y is absent, and the compound of formula (1) is selected from the compounds shown in formula (1L):

[0060]

[0061] In a preferred embodiment, the compound of formula (1) is selected from the compounds represented by formula (1M):

[0062]

[0063] In a preferred embodiment, the compound of formula (1) is selected from:

[0064]

[0065]

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[0140]

[0141] Secondly, the present invention provides a method for preparing the aforementioned ionizable cationic lipid compound, comprising the step of reacting a compound of formula (II) with a compound of formula (III):

[0142]

[0143] Wherein, Xa and Xb are groups containing leaving groups or nucleophilic groups, and Xa and Xb form L1 through nucleophilic reactions or condensation reactions.

[0144] Thirdly, the present invention provides a method for preparing the aforementioned ionizable cationic lipid compound, comprising the step of reacting a compound of formula (IV) with a compound of formula (V):

[0145]

[0146] Xc and Xd are groups containing leaving groups or nucleophilic groups, and Xc and Xd form L2 through nucleophilic reactions or condensation reactions.

[0147] Fourthly, the present invention provides a method for preparing the aforementioned ionizable cationic lipid compound, comprising the step of reacting the compound of formula (VI):

[0148]

[0149] Where Xe is a group containing a leaving group or a nucleophilic group, Xf is a compound containing a leaving group or a nucleophilic group, and Xe and Xf form M through a nucleophilic reaction or a condensation reaction.

[0150] Fifthly, the present invention provides a method for preparing the aforementioned ionizable cationic lipid compound, comprising the steps of reacting compound (VII) sequentially with compound (VIII), compound (IX), and compound (X):

[0151]

[0152] Xg, Xh, Xi, Xj, Xk, and Xl are groups containing leaving groups or nucleophilic groups, and Xg and Xj form G1 through nucleophilic or condensation reactions, Xh and Xk form G2 through nucleophilic or condensation reactions, and Xi and Xl form G3 through nucleophilic or condensation reactions.

[0153] In a sixth aspect, the present invention provides a method for preparing the aforementioned ionizable cationic lipid compound, comprising the step of reacting a compound of formula (XI) with a compound of formula (XII):

[0154] or

[0155] The steps for the reaction of compound (XIII) with compound (XII):

[0156]

[0157] Where Xm is a group containing a nucleophilic group, and Xm has the same C=CL as the compound of formula (XII). 2a X1-L2 is formed through an addition reaction.

[0158] In a preferred embodiment, R1, R2, and R3 are independently groups of the following:

[0159]

[0160] The meanings of Y, R1', and R2' are the same as before;

[0161] This also includes the steps of forming the tail chains R1, R2, and R3:

[0162]

[0163] Where X is a leaving group.

[0164] In a preferred embodiment, the raw materials used in the reaction process also contain protecting groups, and the reaction steps include protection and / or deprotection steps.

[0165] Those skilled in the art will recognize that protecting groups may be necessary to protect certain groups from the effects of reaction conditions. Protecting groups can also be used to distinguish similar functional groups in a molecule. A list of protecting groups and methods for introducing and removing these groups can be found in Wuts, PGM, Greene, TW, Greene's Protective Groups in Organic Synthesis, 4th Edition, John Wiley & Sons: New Jersey, 2007.

[0166] Preferred protecting groups include, but are not limited to: protecting groups for hydroxyl groups, protecting groups for amino or amine groups, protecting groups for carboxyl groups, protecting groups for aldehydes or ketones, protecting groups for thiol groups, and protecting groups for any combination of functional groups such as hydroxyl, amino or amine, carboxyl, aldehyde or ketone, and thiol.

[0167] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the methyl and substituted methyl groups can be selected from: methoxymethyl, meththiomethyl(phenyldimethylsilane)methoxymethyl, benzyloxymethyl, p-methoxybenzyloxymethyl, [(3,4-dimethoxybenzyl)oxy]methyl, p-nitrobenzyloxymethyl, o-nitrobenzyloxymethyl, [(R)-1-(2-nitrophenyl)ethoxy]methyl, (4-methoxyphenoxy)methyl, o-methoxyphenoxymethyl, p-phenylphenoxymethyl, tert-butoxymethyl, 4-isopentoxymethyl, silanoxymethyl, 2-methoxyethoxymethyl, 2-cyanoethoxymethyl, di(2-chloroethoxy)methyl, 2,2,2-trichloroethoxymethyl 2-(trimethylsilyl)ethoxymethyl, mentholoxymethyl, O-di(2-acetoxyethoxy)methyl, 2-tetrahydropyranyl, fluorotetrahydropyranyl, 3-bromo-2-tetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxycyclohexyl, 4-methoxy-tetrahydropyran-4-yl, 4-methoxy-thiotetrahydropyran-4-yl, 4-methoxy-thiotetrahydropyran-4-yl, 1-[ [2-Chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl, 1-(4-chlorophenyl)-4-methoxypiperidin-4-yl, 1,4-dioxane-2-yl, tetrahydrofuran-2-yl, thiotetrahydrofuran-2-yl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-mentholfuran-2-yl.

[0168] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the substituted ethyl group can be selected from: 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-hydroxyethyl, 2-bromoethyl, 1-[2-(trimethylsilane)ethoxy]ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 1-methyl-1-phenoxyethyl, 2,2,2-Trichloroethyl, 1,1-di-p-methoxyphenyl-2,2,2-trichloroethyl, 1,1,1,3,3,3-hexafluoro-2-phenylisopropyl, 1-(2-cyanoethoxy)ethyl, 2-trimethylsilylethyl, 2-(benzylthiol)ethyl, 2-(benzylselenoyl)ethyl, tert-butyl, cyclohexyl, 1-methyl-1'-cyclopropylmethyl, allyl, isopentenyl, cinnamyl, 2-styryl, propargyl, p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, 2,4-dinitrophenyl, 2,3,5,6-tetrafluoro-4-(trifluoromethyl)phenyl, etc.

[0169] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the benzyl and methoxy-substituted benzyl groups can be selected from: p-methoxybenzyl, 3,4-dimethoxybenzyl, 2,6-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, pentadienylnitrobenzyl, pentadienylnitropiperolyl, 2,6-dichlorobenzyl, 2,4-dichlorobenzyl, 2,6-difluorobenzyl, p-cyanobenzyl, fluorobenzyl, or 4-fluoroalkoxybenzyl. p-Trimethylsilylmethylbenzyl, p-Phenylbenzyl, 2-Phenyl-2-propyl, p-P-acetaminobenzyl, p-4-azidobenzyl, 4-azido-3-chlorobenzyl, 2-Trifluoromethylbenzyl, 4-Trifluoromethylbenzyl, p-(methylsulfinyl)benzyl, p-(Si-methyl-cyclobutyryl)benzyl, 4-acetoxybenzyl, 4-(2-trimethylsilyl)ethoxymethoxybenzyl, 2-naphthylmethyl, 2-pyridinemethyl, 4-pyridinemethyl, 3-methyl-N-oxo-2-pyridinemethyl, 2-quinolinemethyl, 6-methoxy-2-(4-methylphenyl)-4-quinoline, 1 -Pyrene methyl, diphenylmethyl, 4-methoxydiphenylmethyl, 4-phenyldiphenylmethyl, di(p-nitrophenyl)methyl, 5-diphenylcycloheptyl, triphenylmethyl, tris(4-tert-butylphenyl)methyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tris(p-methoxyphenyl)methyl, 4-(4'-benzoyl)phenyldiphenylmethyl, 4,4',4”-tris(4,5-dichlorophthalamidophenyl)methyl, 4,4',4”-tris(acetylpropionylphenyl)methyl, 4,4',4”-tris(benzyloxyphenyl)methyl 4,4'-dimethoxy-3”-[N-(imidazolylmethyl)]triphenylmethyl, 4,4'-dimethoxy-3”-[N-(imidazolylmethyl)aminoacyltriphenylmethyl, di(4-methoxyphenyl)-1'-pyrenemethyl, 4-(17-tetrabenzo[a,c,g,i]fluorenylmethyl)-4,4”-dimethoxytriphenylmethyl, 9-anthrayl, 9-(9-phenyl)oxanthracene, 9-phenylthioxanthracene, 9-(9-phenyl-10-oxo)anthrayl, 4,5-di(ethoxycarbonyl)-[1,3]-dioxopentane-2-yl, S,S-dioxobenzisothiazole, etc.

[0170] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the silyl group can be selected from: trimethylsilyl, triethylsilyl, triisopropylsilyl, dimethylisopropylsilyl, diethylisopropylsilyl, dimethyl-n-hexylsilyl, 2-norcanane dimethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl. Triphenylsilyl, tri-p-methylphenylsilyl, triphenylsilyl, diphenylmethyl, di-tert-butylmethylsilyl, di(tert-butyl)-1-pyrene methoxysilyl, tri(trimethylsilyl)silyl, (2-hydroxystyryl)dimethylsilyl, (2-hydroxystyryl)diisopropylsilyl, tert-butylmethoxyphenylsilyl, tert-butoxydiphenylsilyl, 1,1,3,3-tetraisopropyl-3-[2-(triphenylmethoxy)ethoxy]disiloxane-1-yl, fluoroalkylsilyl, etc.

[0171] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the acyl group can form an ester with an alcohol, which can be: formate, benzyloxyformate, ethyl ester, chloroacetate, dichloroacetate, triacetate, trichloroacetylimine ester, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, phenylacetate, p-polymer phenylacetate, diphenylacetate, 3-phenyl Propionates, propionates containing two fluorinated alkyl chains, 4-pentenoate, fructose acid, 4,4-(ethylene dithio)pentanoate, 5-[3-di(4-methoxyphenyl)hydroxymethylphenoxy]fructate, neopentanoate, 1-adamantanecarboxylic acid, 2-butenoate, 4-methoxy-2-butenoate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate, 4-bromobenzoate, 2,5-difluorobenzoate, p-nitrobenzoate, pyridine-2-carboxylate, nicotinic acid, 2-(azidomethyl)benzoate, 4-azidobutyrate, (2-azidomethyl)benzoate 2-{[(triphenylmethyl mercapto)oxy]methyl}benzoate, 2-{[(4-methoxytriphenylmethyl mercapto)oxy]methyl}benzoate, 2-{[methyl(triphenylmethyl)amino]methyl}benzoate, 2-{{[(4-methoxytriphenylmethyl) mercapto]methylamino}-methyl}benzoate, 2-(allyloxy)phenylacetate, 2-(isopentenoxymethyl)benzoate, 6-(fructoyloxymethyl)-3-methoxy-2-nitrobenzoate, 6-(fructoyloxymethyl)-3-methoxy-4-nitrobenzoate, 4 2-Benzyloxybutyrate, 4-triallylsiloxybutyrate, 4-acetoxy-2,2-dimethylbutyrate, 2,2-dimethyl-4-valerate, 2-iodobenzoate, 4-nitro-4-methylvalerate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 4-(meththiomethoxy)butyrate, 2-(meththiomethoxy)benzoate, 2-(chloroacetoxymethyl)benzoate, 2-[(2-chloroacetoxy)ethyl]benzoate, 2-[2-(benzyloxy)ethyl]benzoate, 2-[2-(4-methoxybenzyloxy)ethyl]benzoate, etc.

[0172] In a preferred embodiment, the protecting group for the hydroxyl group may be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the sulfonate, sulfinate, or sulfenate is, for example, sulfate, allyl sulfonate, methanesulfonate, benzyl sulfonate, p-toluenesulfonate, 2-[(4-nitrophenyl)ethyl]sulfonate, 2-trifluoromethylsulfonate, 4-monomethoxytriphenylmethyl sulfenate, 2,4-dinitrophenyl sulfenate, 2,2,5,5-tetramethylpyrrolidine-3-ton-1-sulfinate, etc.

[0173] In a preferred embodiment, the protecting group for the hydroxyl group may be a borate ester or a dimethyl thiophosphate ester.

[0174] In a preferred embodiment, the protecting group for the hydroxyl group can be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the carbonate can be: methyl carbonate, methoxymethyl carbonate, 9-fluorenylmethyl carbonate, ethyl carbonate, bromoethyl carbonate, 2-(meththiomethoxy)ethyl, 2,2,2-trichloroethyl carbonate, 1,1-dimethyl- 2,2,2-Trichloroethyl carbonate, 2-(trimethylsilyl)ethyl carbonate, 2-[dimethyl(2-naphthylmethyl)silyl]ethyl carbonate, 2-(benzenesulfonyl)ethyl carbonate, 2-(triphenylphosphino)ethyl carbonate, Cis-[4-[[(-methoxytriphenylmethyl)sulfenic acid]oxy]tetrahydrofuran-3-yl]oxy carbonate, isobutyl carbonate, tert-butyl carbonate, vinyl carbonate, allyl carbonate, cinnamyl carbonate, Propylene carbonate, p-chlorophenyl carbonate, p-nitrophenyl carbonate, 4-ethoxy-1-naphthyl carbonate, 6-bromo-7-hydroxycoumarin-4-ylmethyl carbonate, benzyl carbonate, o-nitrobenzyl carbonate, p-nitrobenzyl carbonate, p-methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, anthraquinone-2-ylmethyl carbonate, 2-dansylethyl carbonate, 2-(4-nitrophenyl)ethyl carbonate, 2-(2,4-dinitrophenyl)ethyl carbonate Esters, 2-(2-nitrophenyl)propyl carbonate, 2-(3,4-methylenedioxy-6-nitrophenylpropyl 294 carbonate, 2-cyano-1-phenylethyl carbonate, 2-(2-pyridyl)amino-1-phenylethyl carbonate, 2-[N-methyl-N-(2-pyridyl)]amino-1-phenylethyl carbonate, benzoylmethyl carbonate, 3',5'-dimethoxybenzoyl carbonate, methyl dithiocarbonate, S-benzyl thiocarbonate, etc.;

[0175] In a preferred embodiment, the protecting group for the hydroxyl group may be a substituted methyl, substituted ethyl, substituted benzyl, silyl, acyl, sulfonyl, sulfinyl, sulfenyl, carbamoyl, carbonate, borate, or phosphoryl group, wherein the carbamoyl may be N,N-dimethylthiocarbamate, N-phenylcarbamate, N-methyl-N-(o-nitrophenyl)carbamate, etc.

[0176] In a preferred embodiment, 1,2-diol or 1,3-diol can be protected by forming a cyclic acetal or ketal, a cyclic orthoester, a silane derivative, a cyclic carbonate, or a cyclic borate ester. The protecting group forming the cyclic acetal or ketal can be: methylene, ethylene, tert-butylmethylene, 1-tert-butylethylene, 1-phenylethylene, 2-(methoxycarbonyl)ethylene, 2-(tert-butylcarbonyl)ethylene, benzenesulfonylethylene, 2,2,2-trichloroethylene, 3-(benzyloxy)propylene, acrolein acetal, or acetone (isopropylene). Cyclopentanediol, cyclohexyldiol, cycloheptanediol, benzyldiol, p-methoxybenzyldiol, 1-(4-methoxyphenyl)ethylene, 2,4-dimethoxybenzyldiol, 3,4-dimethoxybenzyldiol, p-acetoxybenzyldiol, 4-(tert-butyldimethylsiloxy)benzyldiol, 2-nitrobenzyldiol, 4-nitrobenzyldiol, mesitylene, 6-bromo-7-hydroxycoumarin-2-methylene, 1-naphthaldehyde acetal, 2-naphthaldehyde acetal, 9-anthraethyl acetal, benzophenone ketal, di-(p-anesinyl)methylene ketal, xanthene-9-ene, 2,7-dimethylxanthene-9-ene.

[0177] In a preferred embodiment, 1,2-diol or 1,3-diol can be protected by forming a cyclic acetal or ketal, a cyclic orthoester, a silane derivative, a cyclic carbonate, or a cyclic borate, wherein the protecting group forming the cyclic orthoester can be: methoxymethylene, ethoxymethylene, 2-oxacyclopentanediol, dimethoxymethylene, 1-methoxyethylene, 1-ethoxyethylene, methylene, phthalide, 1,2-dimethoxyethylene, α-methoxybenzylene, 1-(N,N-dimethylamino)ethylene derivative, α-(N,N-dimethylamino)benzylene derivative, butane-2,3-bisacetal, cyclohexane-1,2-diacetal, dispirol ketal, etc.

[0178] In a preferred embodiment, the 1,2-diol or 1,3-diol can be protected by forming a cyclic acetal or ketal, a cyclic orthoester, a silane derivative, a cyclic carbonate, or a cyclic borate, wherein the silane derivative protecting group can be: di-tert-butylsilenyl, dialkylsilenyl, 1,3-(1,1,3,3-tetraisopropyldisiloxane) derivative, 1,1,3,3-tetra-tert-butoxydisiloxane derivative, methylene-bis-(diisopropylsiloxane)methylene, 1,1,4,4-tetraphenyl-1,4-disiloxane, o-xylene ether, 3,3'-oxybis(dimethoxytriphenylmethyl) ether, 1,2-ethylene-3,3-bis(4”4'-dimethoxytriphenylmethyl) ether.

[0179] In a preferred embodiment, the 1,2-diol or 1,3-diol can be protected by forming a cyclic acetal or ketal, a cyclic orthoester, a silane derivative, a cyclic carbonate, or a cyclic borate, wherein the protecting group forming the cyclic carbonate can be a formyl group.

[0180] In a preferred embodiment, the 1,2-diol or 1,3-diol can be protected by forming a cyclic acetal or ketal, a cyclic orthoester, a silane derivative, a cyclic carbonate, or a cyclic borate, wherein the protecting group forming the cyclic borate can be methyl borate, ethyl borate, phenyl borate, o-acetaminophenylboronic acid ester, etc.

[0181] In a preferred embodiment, the protecting group for the hydroxyl group is preferably selected from the following protecting groups: tetrahydropyranyl (THP), methyl (Me), benzyl (Bn), methoxymethyl (MOM), allyl (All), triphenylmethyl (Trt), acetyl (Ac), pivaloyl (Piv), tert-butyldimethylsilyl (TBDMS), tert-butyldimethylsilyl (TBDPS), trimethylsilyl (TMS), triethylsilyl (TES), phthalimide (Phth), tert-butyloxycarboxylate (Boc), benzyloxycarboxylate (Cbz), and 9-fluorenemethoxycarboxylate (Fmoc).

[0182] In a preferred embodiment, the protecting group for the carboxylic acid may be a substituted methyl ester, a substituted ethyl ester, a phenyl ester, a substituted phenyl ester, a silyl ester, an amino group, or a hydrazine group, wherein the substituted methyl ester may be: 9-fluorenyl methyl ester, methoxymethyl ester, 2-methoxyethoxymethyl ester, meththiomethyl ester, 2-tetrahydropyran ester, 2-tetrahydrofuran ester, 2-(trimethylsilyl)ethoxymethyl ester, benzyloxymethyl ester, triisopropylsiloxymethyl ester, tervamoloxymethyl ester, etc. Phenylacetoxymethyl ester, triisopropylsilyl methyl ester, cyanomethyl ester, acetone alcohol ester, benzoyl methyl ester, p-bromobenzoyl methyl ester, α-methylbenzoyl methyl ester, p-methoxybenzoyl methyl ester, 3,4,5-trimethoxybenzoyl methyl ester, 2,5-dimethylbenzoyl methyl ester, phenylethyl ketone ester, aminoacyl methyl ester, p-azoanilinecarbamoyl methyl ester, 6-bromo-7-hydroxycoumarin-4-ylmethyl ester, N-phthalamide methyl ester, etc.

[0183] In a preferred embodiment, the protecting group for the carboxylic acid is a substituted ethyl ester, such as: 2,2,2-trichloroethyl ester, 2-haloethyl ester, A-chloroalkyl ester, 2-(trimethylsilyl)ethyl ester, (2-methyl-2-trimethylsilyl)ethyl ester, (2-phenyl-2-trimethylsilyl)ethyl ester, 2-methylthioethyl ester, 1,3-dithiazide-2-methyl ester, 2-(p-nitrophenylsulfinyl)ethyl ester, 2-(p-methylbenzenesulfonyl)ethyl ester, 2-(2'-pyridyl)ethyl ester, 2-(diphenylphosphino)ethyl ester, (p-methoxy)ethyl ester, etc. 1-Methyl-1-phenylethyl ester, 2-(4-acetyl-2-nitrophenyl)ethyl ester, 1-[2-(2-hydroxyalkyl)phenyl]ethyl ketone, 2-cyanoethyl ester, tert-butyl ester, 3-methyl-3-pentyl ester, dicyclopropylmethyl ester, 2,4-dimethyl-3-pentyl ester, cyclopentyl ester, cyclohexyl ester, allyl ester, methyl allyl ester, 2-methyl-3-buten-2-yl ester, 3-methyl-2-butenyl ester, 3-butenyl ester, 4-(trimethylsilyl)-2-buten-1-yl ester, cinnamic acid ester, α-methyl cinnamic acid ester, propyne ester, etc.

[0184] In a preferred embodiment, the protecting group for the carboxylic acid is a phenyl ester or a substituted phenyl ester, such as: phenyl ester, 2,6-dimethylphenyl ester, 2,6-diisopropylphenyl ester, 2,6-di-tert-butyl-4-methylphenyl ester, 2,6-di-tert-butyl-4-methoxyphenyl ester, p-(dimethylthio)phenyl ester, pentafluorophenyl ester, 2-(dimethylamino)-5-nitrophenyl ester, etc.;

[0185] In a preferred embodiment, the protecting group for the carboxylic acid is a benzyl ester or a substituted phenyl ester, such as: benzyl ester, triphenylmethyl ester, 2-chlorophenyldiphenylmethyl ester, 2,3,4,4',4”,5,6-heptafluorotriphenylmethyl ester, diphenylmethyl ester, di(o-nitrophenyl)methyl, 9-anthraylmethyl ester, 2-(9,10-dioxo)anthraylmethyl ester, 5-dibenzocycloheptanyl ester, 1-pyrene methyl ester, 2-(trifluoromethyl)-6-cryoglycol methyl ester, 2,4,6-trimethylbenzyl ester, p-bromobenzyl ester, o-nitrobenzyl ester, p-nitrobenzyl ester, p-methoxy Benzyl esters, 2,6-dimethoxybenzyl esters, 4-(methylhypoxanyl)benzyl esters, 4-sulfonic acid benzyl esters, 4-azidomethoxybenzyl esters, 4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylene dimethyl)-3-methylbutyl]amino}benzyl esters, piperyl esters, 4-pyridine methyl esters, polymer-substituted benzyl esters, 2-naphthyl methyl esters, 3-nitro-2-naphthyl methyl esters, 4-quinolinyl methyl esters, 8-bromo-7-hydroxyquinoline-2-yl methyl esters, 2-nitro-4,5-dimethoxybenzyl esters, 1,2,3,4-tetrahydro-1-naphthyl esters, etc.

[0186] In a preferred embodiment, the protecting group for the carboxylic acid may be a silyl ester, such as: trimethylsilyl ester, tri- and silyl ester, tert-butyl dimethylsilyl ester, tert-butyl diphenylsilyl ester, isopropyl dimethylsilyl ester, phenyl dimethylsilyl ester, di-tert-butyl methylsilyl ester, triisopropylsilyl ester, tri(2,6-diphenylbenzyl)silyl ester, etc.

[0187] In a preferred embodiment, the protecting group for the carboxylic acid may be an amino group or a hydrazine group, such as: N,N-dimethylamino, pyrrolidone, piperidinyl, 5,6-dihydrophenanthridine, o-nitroaniline, N-7-nitroindolyl, N-8-nitro-1,2,3,4-tetrahydroquinolinyl, 2-(2-aminophenyl)acetaldehyde dimethyl acetal, p-polymer-benzenesulfonamide, N-phenylhydrazine, N,N'-dimethylhydrazine, N,N'-diisopropylhydrazine, etc.

[0188] In a preferred embodiment, the protection of the carboxylic acid can also be achieved by chemically converting the carboxylic acid group into the following groups: 2-alkyl-1,3-oxazoline, 4-alkyl-5-oxo-1,3-oxazoline, 2,2-ditrifluoromethyl-4-alkyl-5-oxo-1,3-oxazoline, 2,2-dimethyl-4-alkyl-2-silyl-5-oxo-1,3-oxazoline, 2,2-difluoro-1,3,2-oxazoborane-5-one, 5-alkyl-4-oxo-1,3-dioxopentane, dioxocyclohexanone, orthoester, pentaaminocobalt(III) complex, etc.

[0189] In a preferred embodiment, the protecting group for the carboxyl group is preferably selected from the following protecting groups: tert-butyl ester (t-Bu), benzoyl ester (Bn), methyl ester (Me), ethyl ester (Et), allyl ester (All), isopropyl ester (i-Pr), isobutyl ester (i-Bu), triphenylmethyl ester (Trt), methoxymethyl (MOM), tert-butyldimethylsilane ester (TBDMS), and 9-fluorenylmethoxycarbonyl (Fmoc).

[0190] In a preferred embodiment, the protecting group for the amino or amine group may be a carbamate ROC(O)N(R')2, where N(R')2 is the protected group, and R may be a methyl or substituted methyl group, such as: methyl, 9-fluorenylmethyl, 2,6-di-tert-butyl-9-fluorenylmethyl, 2,7-di(trimethylsilyl)fluorenylmethyl, 9-(2-sulfonyl)fluorenylmethyl, 9-(2,7-di-... (Brominated)fluorenylmethyl, 17-tetrabenzo[a,c,g,i]fluorenylmethyl, 2-chloro-3-indenemethyl, benzo[f]indene-3-ylmethyl, 1,1-dioxobenzo[b]thiophene-2-ylmethyl, 2-methanesulfonyl-3-phenyl-1-prop-2-enoyloxy, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthoneyl)]methyl, etc.

[0191] In a preferred embodiment, the protecting group for the amino or amine group may be a carbamate ROC(O)N(R')2, where N(R')2 is the protected group, and R may be an ethyl or substituted ethyl group, such as: 2,2,2-trichloroethyl, 2-trimethylsilylethyl, (2-phenyl-2-trimethylsilyl)ethyl, 2-phenylethyl, 2-chloroethyl, 1,1-dimethyl-2-haloethyl, 1,1-dimethyl-2,2-dibromoethyl, 1,1-dimethyl-2,2,2-trichloroethyl, 2-(2'-pyridyl)ethyl, 2-(4'-pyridyl)ethyl, 2,2-di(4'-nitrophenyl)ethyl, 2-[(2-nitrophenyl)dimercapto]-1-phenylethyl, 2-(N,N-dicyclohexylcarbamoyl)ethyl, tert-butyl, Fluorinated Boc, 1-adamantyl, 2-adamantyl, 1-(1-adamantane)-1-methylethyl, 1-methyl-1-(4-biphenyl)ethyl, 1-(3,5-di-tert-butylphenyl)-1-methylethyl, triisopropylsilyl, vinyl, allyl, isopentenyl, 1-isopropylallyl, cinnamyl, 4-nitrocinnamyl, 3-(3'-pyridyl)prop-2-enyl, hexadienyl, propargyl, but-2-yn-1,4-diyl, 8-quinolinyl, N-hydroxypiperidinyl, alkyl dithioyl, benzyl, 3,5-di-tert-butylbenzyl, p-methoxybenzyl, p-nitrobenzyl, p-bromobenzyl, p-chlorobenzyl, 2,4-dichlorobenzyl, 4-methylsulfonyluylbenzyl, 4-trifluoromethylbenzyl, fluorobenzyl, 2-naphthylmethyl, 9-anthraylmethyl, diphenylmethyl, etc.

[0192] In a preferred embodiment, the protecting group for the amino or amine group can be a carbamate ROC(O)N(R')2, where N(R')2 is the protected group, and R can be one of the following groups: 4-phenylacetylbenzyl, 4-azidobenzyl, 4-azidomethoxybenzyl, m-chloro-p-acetyloxybenzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolylmethyl, 2-(trifluoromethyl)-6-cryomethyl, 2-methylthioethyl, 2-methanesulfonylethyl, 2-(p-toluenesulfonyl)ethyl, 2-(4-nitrobenzenesulfonyl)ethyl, 2-(2,4-dinitrobenzenesulfonyl)ethyl, etc. Acyl)ethyl, 2-(4-trifluoromethylbenzenesulfonyl)ethyl, [2-(1,3-dithiaalkyl)]methyl, 2-alkylphosphineethyl, 2-[phenyl(methyl)thioether]ethyl, 1-methyl-1-(triphenylphosphine)ethyl, 1,1-dimethyl-2-cyanoethyl, 2-dansylethyl, 2-(4-nitrophenyl)ethyl, 4-methylthiophenyl, 2,4-dimethylthiophenyl, m-nitrophenyl, 3,5-dimethoxybenzyl, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl, α-methylnitropiperyl, nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl 3,4-Disubstituted-6-nitrobenzyl, phenyl(o-nitrophenyl)methyl, 2-nitrophenylethyl, 6-nitroveratrol, 4-methoxybenzoylmethyl, 3',5'-dimethoxybenzoyl, 9-xanthonylmethyl, N-methyl-N-(o-nitrophenyl), N-(2-acetoxyethyl)amino, 1-methyl-1-cyclopropylmethyl, tert-amyl, 1-methylcyclobutyl, 1-methylcyclohexyl, cyclobutyl, cyclopentyl, cyclohexyl, isobutyl, isoborneol, cyclopropylmethyl, p-decoxybenzyl, diisopropylmethyl, 2,2-dimethoxycarbonylvinyl, o-(N,N-di) Benzyl methylformamide, 1,1-dimethyl-3-(N,N-dimethylformamide)propyl, butynyl, 1,1-dimethylpropynyl, 2-iodoethyl, 1-methyl-1-(4'-pyridyl)ethyl, 1-methyl-1-(p-phenylazophenyl)ethyl, p-(p-methoxyphenylazo)benzyl, p-(phenylazo)benzyl, 2,4,6-trimethylbenzyl, isonicotinic acid, 4-(trimethylammonium)benzyl, p-cyanobenzyl, di(2-pyridyl)methyl, 2-furanylmethyl, phenyl, 2,4,6-tri-tert-butylphenyl, 1-methyl-1-phenylethyl, benzyl thiocarbamate.

[0193] In a preferred embodiment, the protecting group for the amino or amine group can be a urea or thiourea derivative RN(R')2, where N(R')2 is the protected group and R can be selected from the following groups: carbamoyl, phenothiazinyl-(10)-carbonyl derivative, N'-p-toluenesulfonylaminocarbonyl, N'-phenylaminothiocarbonyl, 4-hydroxyphenylaminocarbonyl, 3-hydroxytryptamine carbonyl, N'-phenylaminothiocarbonyl, etc.

[0194] In a preferred embodiment, the protecting group for the amino or amine group may be amide RN(R')2, wherein N(R')2 is the protected group, and R may be selected from the following groups: formyl, acetyl, chloroacetyl, trichloroacetyl, trifluoroacetyl, phenylacetyl, 3-phenylpropionyl, pent-4-enoyl, pyridylformyl, 3-pyridylformyl, N-benzoylphenylpropane derivative, benzoyl, p-phenylbenzoyl, o-nitrophenylacetyl, 2,2-dimethyl-2-o-nitrophenylacetyl, o-nitrophenoxyacetyl, 3-(o-nitrophenyl)propionyl, 2-methyl-2-(o-nitrophenoxy)propionyl, 3-methyl-3-nitrobutyryl, o-Nitrocinnamyl, o-Nitrobenzoyl, 3-(4-tert-butyl-2,6-dinitrophenyl)-2,2-dimethylpropionyl, o-(benzoyloxymethyl)benzoyl, 2-(acetoxymethyl)benzoyl, 2-[(tert-butyldiphenylsiloxy)methyl]benzoyl, 3-(3',6'-dioxo-2',4',5-trimethylcyclohexyl-1,4-diene)-3,3-dimethylpropionyl, o-hydroxytrans-cinnamyl, 2-methyl-2-(o-phenylazophenoxy)propionyl, 4-chlorobutyryl, acetylacetyl, 3-(p-hydroxyphenyl)propionyl, (N'-dithiobenzyloxycarbonylamino)acetyl, N-acetylated methionine derivatives, etc.

[0195] In a preferred embodiment, the primary amino group may be double-protected, and the protecting group may be selected from the following groups: 4,5-diphenyl-3-oxazoline-2-one, N-phthaloyl, N-dichlorophthaloyl, N-tetrachlorophthaloyl, N-4-nitrophthaloyl, N-thiodiglycol, N-dithiosuccinoyl, N-2,3-diphenylmaleyl, N-2,3-dimethylmaleyl, N-2,5-dimethylpyrrole, N-2,5- Bis(triisopropylsiloxy)pyrrole, N-1,1,4,4-tetramethyl-1,4-disil-5azacyclopentane, N-1,1,3,3-tetramethyl-1,3-disilisoindoline, N-diphenylsilyldiethylene group, N-5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, N-5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridone, 1,3,5-dioxazine.

[0196] In a preferred embodiment, for N-alkyl-substituted and N-aryl-substituted amino groups, the protecting group may be selected from the following groups: N-methyl, N-tert-butyl, N-allyl, N-isopreneyl, N-cinnamyl, N-phenylallyl, N-propynyl, N-methoxymethyl, N-[2-(trimethylsilyl)ethoxy]methyl, N-3-acetoxypropyl, N-cyanomethyl, 2-azanorbornene, N-2,4-dinitrophenyl, o-methoxy or p-methoxyphenyl N-Benzyl, N-4-Methoxybenzyl, N-2,4-Dimethoxybenzyl, N-2-Hydroxybenzyl, N-9-Phenylofluoroenyl, N-fluorenyl, N-Ferrocenemethyl, N-2-Methylpyridinium N'-oxide, N-7-Methoxy-4-ylmethyl, N-Diphenylmethyl, N-Bis(4-Methoxyphenylmethyl), N-5-Dibenzophenylethyl, N-Triphenylmethyl, N-(4-Methylphenyl)diphenylmethyl, N-(4-Methoxyphenyl)diphenylmethyl, etc.

[0197] In a preferred embodiment, the protection of the amino or amine group can be achieved by preparing an imine derivative, such as: N-1,1-dimethylthiomethylene, N-phenylmethylene, N-p-methoxyphenylmethylene, N-diphenylmethylene, N-[(2-pyridyl)methylene]methylene, N-(N',N'-dimethylaminomethylene), N-(N',N'-dibenzylaminomethylene), N-(N'-tert-butylaminomethylene), N,N'-isopropylidene, N-p-nitrophenylmethylene, N-salicylyl, N-5-chlorosalicylyl, N-(5-chloro-2-hydroxyphenyl)phenylmethylene, N-cyclohexylene, N-tert-butylmethylene, etc.

[0198] In a preferred embodiment, the protection of the amino or amine group can be achieved by preparing an enamine derivative, such as: N-(5,5-dimethyl-3-oxo-1-cyclohexenyl), N-2,7-dichloro-9-fluorenylmethylene, N-1-(4,4-dimethyl-2,6-dioxocyclohexenyl)ethyl, N-(1,3-dimethyl-2,4,6-(1H,3H,5H)-trioxopyrimidin-5-yl)methyl, N-4,4,4-trifluoro-3-oxo-1-butenyl, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrololin-3-yl).

[0199] In a preferred embodiment, the amino or amine group can be protected by derivatives formed by the reaction of the N atom of the amino or amine group with other heteroatoms (e.g., N, O, P, Si, or S), such as: N-nitro, N-nitroso, N-oxide, azide, triazacycloene, N-trimethylsilylmethyl-N-benzylhydrazine, N-diphenylphosphineamide, dimethyl and diphenylthiophosphineamide, dialkylphosphineamide, dibenzyl and diphenylphosphineamide, iminotriphenylphosphine, triphenylmethylsilyl, benzylthioamide, 2-nitrobenzenethioamide, 2,4-dinitrobenzenethioamide, pentachlorobenzenethioamide, 2-nitro-4-methoxybenzenethioamide, triphenylmethylthioamide, 1-(2,2,2-trifluoro-1,1-diphenyl)ethylthioamide, N-3-nitro-2-pyridylthioamide, methanesulfonamide, trifluorosulfonamide, tert-butylsulfonamide, benzylsulfonamide, 2-(trimethylsilyl) Ethylsulfonamide, p-toluenesulfonamide, benzenesulfonamide, anisylsulfonamide, 2- or 4-nitrobenzenesulfonamide, 2,4-dinitrobenzenesulfonamide, 2-naphthalenesulfonamide, 4-(4',8'-dimethoxynaphthyl)benzenesulfonamide, 2-(4-methylphenyl)-6-methoxy-4-methylsulfonamide, 9-anthraphenenesulfonamide, pyridine-2-sulfonamide, benzothiazole-2-sulfonamide, benzoylmethylsulfonamide, 2,3,6-trimethyl-4-methyl Oxybenzenesulfonamide, 2,4,6-trimethoxybenzenesulfonamide, 2,6-dimethyl-4-methoxybenzenesulfonamide, pentamethylbenzenesulfonamide, 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide, 4-methoxybenzenesulfonamide, 2,4,6-trimethylbenzenesulfonamide, 2,6-dimethoxy-4-methylbenzenesulfonamide, 3-methoxy-4-tert-butylbenzenesulfonamide, 2,2,5,7,8-pentamethylbenzopyran-6-sulfonamide, etc.

[0200] In a preferred embodiment, protection of amino alcohols or amino groups can be achieved by reacting an amino or amino group with a hydroxyl group to a 2-azolidinone or oxazoline.

[0201] In a preferred embodiment, the nitrogen-containing aromatic heterocycle may be selected from the N-sulfonyl derivatives, carbamates, N-alkyl and N-aryl derivatives, N-trialkylsilyl, N-allyl, N-benzyl, amides, amino acetal derivatives, and protecting groups of amides and xanthines as described above.

[0202] In a preferred embodiment, the protecting group for the amine group is preferably selected from the following protecting groups: 9-fluorenylmethoxycarbonyl (Fmoc), benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), trichloroethoxycarbonyl (Troc), allyloxycarbonyl (Alloc), methoxycarbonyl (Moc), acetyl (Ac), trifluoroacetyl (TFA), 2,4-dimethoxybenzyl (DMB), benzyl (Bn), triphenylmethyl (Tr), benzyloxymethyl (Bom), p-toluenesulfonyl (Ts), 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr), phthalimide (Phth), and benzylmethylamine.

[0203] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating an acetal or ketal, a dithioacetal or monothioacetal or ketal, a nitrile, a substituted hydrazone, an oxime, a 1,2-adduct, or by converting the carbonyl group to an enol anion, an enol ester, an enamine, or an imine.

[0204] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating an acetal or ketal, wherein the protecting group can be selected from: dimethyl, diisopropyl, bis(2,2,2-trichloroethyl), dibenzyl, bis(2-nitrobenzyl), diacetyl, 1,3-dioxane, 5-methylene-1,3-dioxane, 5-trimethylsilyl-1,3-dioxane, 5,5-dibromo-1,3-dioxane, 5-(2-pyridyl)-1,3-dioxane, salicylic acid acetal. 1,3-Dioxolane, 4,4,5,5-Tetramethyl-1,3-Dioxolane, 4-Bromomethyl-1,3-Dioxolane, 4-Phenylenylmethyl-1,3-Dioxolane, 4-(3-Butenyl)-1,3-Dioxolane, 4-Phenyl-1,3-Dioxolane, 4-(4-Methoxyphenyl)-1,3-Dioxolane, 4-(2-Nitrophenyl)-1,3-Dioxolane, 4-(4-Nitrophenyl)-1,3-Dioxolane, 4-Fluoro-1,3-Dioxolane, 4 -[6-bromo-7-hydroxycoumarin-4-yl]-1,3-dioxane, 4-trimethylsilyl-1,3-dioxolane, O,O'-phenyldioxyketal, 1,3-dioxane, 1,5-dihydro-3H-2,4-benzodioxane, 7,7-dimethyl-1,2,4-trioxane, 3,3-dialkyl-6-(1-phenylvinyl)-1,2,4-trioxane, (4R,5R)-diphenyl-1,3-dioxolane, 4,5-dimethyl... Examples of its compounds include methyl-1,3-dioxolane, trans-1,2-cyclohexanediol ketal, trans-4,6-dimethyl-1,3-dioxane, 4,5-bis(dimethylaminocarbonyl)-1,3-dioxolane, 4,5-dicarboxymethyl-1,3-dioxolane, 4,5-dimethoxymethyl-1,3-dioxolane, 2,2-dialkyl-4,5-bis(2-nitrophenyl)-1,3-dioxolane, and 4,5-bis(2-nitro-4,5-dimethoxyphenyl)-1,3-dioxolane.

[0205] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating a dithioacetal or ketal, wherein the protecting group can be selected from: S,S'-dimethyl, S,S'-diethyl, S,S'-dipropyl, S,S'-dibutyl, S,S'-dipentyl, S,S'-diphenyl, S,S'-dibenzyl, S,S'-diacetyl, 1,3-dithiaran, 1,3-dithiocycloalkanes, 1,5-dihydro-3H-2,4-dithionecycloheptane, etc.

[0206] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating a monothioacetal or ketal, wherein the protecting group can be selected from: O-trimethylsilyl-S-alkyl, O-alkyl-S-alkyl or -S-phenyl, O-methyl-S-2-methylthioethyl, 1,3-oxothiapentane, etc.

[0207] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating an O-substituted geminitrogen, wherein the protecting group can be selected from: O-acetyl, O-methoxycarbonyl, O-trimethylsilyl, O-1-ethoxyethyl, O-tetrahydropyranyl, etc.

[0208] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating a substituted hydrazone, the protecting group being selected from: N,N-dimethyl, phenyl, 2,4-dinitrophenyl, p-toluenesulfonyl, hemicarbazone, diphenylmethyl, etc.

[0209] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by generating a substituted oxime, the protecting group being selected from: O-methyl, O-benzyl, or O-thiophene methyl, etc.

[0210] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by forming a 1,2-adduct, wherein the protecting group can be selected from: diethylamine adduct, N-methoxy-N-methylamine adduct, pyrrole adduct, 1-methyl-2-(1'-hydroxyalkyl)imidazolium, O-silylimidazoliumamine acetal, sodium bisulfite adduct, O-carborane, aminobutyronitrile derivative, N,N'-dimethylimidazolium, N,N'-diarylimidazolium, and 2,3-dihydro-1,3-benzothiazole.

[0211] In a preferred embodiment, the protection of the aldehyde or ketone group can be achieved by converting the carbonyl group into an enol anion, enol ester, enamine, or imine, such as lithium diisopropylamide (LDA), trimethylsilylenol ether, enamine, imine, substituted methylene derivatives, methylaluminum bis(2,6-di-tert-butyl-4-methylphenyloxide) complex, etc.

[0212] In a preferred embodiment, the compound containing a dicarbonyl group may selectively protect one of the carbonyl groups, wherein for α- or β-diketones, the following protections may be selected: enamine, enol acetate, enol ether, methyl, ethyl, isobutyl, methoxyethoxymethyl, methoxymethyl, enamine derivatives, 4-methyl-1,3-dioxolane-enol acetate, pyrrolidine enamine, benzylenol ether, butylthioenol ether, etc.

[0213] In a preferred embodiment, the protecting group for the aldehyde or ketone group is preferably selected from the following protecting groups: O,O-dimethyl acetal (ketone), 1,3-dioxane, 1,3-dioxolane, S,S'-dimethylthioacetal (ketone), 1,3-dithiaane, 1,3-dithionecyclopentane, N,N-dimethylhydrazine (DMH), 2,4-dinitrophenylhydrazine (DNPH), aminourea, oxime, trimethylsilyl (TMS), Tollens' reagent (silver ammonia solution), etc.

[0214] In a preferred embodiment, the protection of the thiol group can be achieved by reacting to form a thioether, a thioester, an asymmetric disulfide bond, or a sulfinyl derivative, wherein the thioether protecting group can be: S-alkyl, S-benzyl, S-p-methoxybenzyl, S-o- or S-p-hydroxy or acetoxybenzyl, S-p- and S-o-nitrobenzyl, S-2,4,6-trimethylbenzyl, S-2,4,6-trimethoxybenzyl, S-4-pyridylmethyl, S-2-methylpyridinyl N-oxide, S-2-quinolinylmethyl, S-9-anthramethyl, S-9-fluorenylmethyl, S-xanthyl, S-ferroceneylmethyl, S-diphenylmethyl, S-bis(4-methoxyphenyl)methyl, S-5-dibenzocycloheptyl, S-triphenylmethyl, 4-methoxytriphenylmethyl, S-diphenyl-4-pyridylmethyl, S-phenyl, S-2,4-dinitrophenyl, S-2-quinolinyl, S-tert-butyl, S-1-adamantyl, S-methoxymethyl, S-isobutoxymethyl S-Benzyloxymethyl, S-1-ethoxyethyl, S-2-tetrahydropyranyl, S-benzylthiomethyl, S-phenylthiomethyl, thiazolidinyl, S-acetaminomethyl, S-trimethylacetaminomethyl, S-benzamidemethyl, S-allyloxycarbonylaminomethyl, N[2,3,5,6-tetrafluoro-4-(N'-piperidinyl)phenyl]-N-allyloxycarbonylaminomethyl, phthalimidemethyl, S-phenylacetaminomethyl, S-acetylmethyl, S-carboxyl Methyl, S-cyanomethyl, S-(2-nitro-1-phenyl)ethyl, S-2-(2,4-dinitrophenyl)ethyl, S-2-(4'-pyridyl)ethyl, S-2-cyanoethyl, S-2-(trimethylsilyl)ethyl, S-2,2-bis(carbonylethoxy)ethyl, S-(1-m-nitrophenyl-2-benzoyl)ethyl, S-2-benzenesulfonylethyl, S-1-(4-methylbenzenesulfonyl)-2-methylpropyl-2-yl, S-p-hydroxybenzoylmethyl.

[0215] In a preferred embodiment, the thiol group can be protected by reacting to form a thioether, a thioester, an asymmetric disulfide bond, or a sulfinyl derivative. The thioester protecting group can be: S-acetyl derivative, S-benzoyl derivative, S-2-methoxyisobutyryl, S-trifluoroacetyl derivative, SN-[[(p-biphenyl)isopropoxy]carbonyl]-N-methyl-γ-aminothiobutyrate, SN-(tert-butoxycarbonyl)-N-methyl-γ-aminothiobutyrate, S-2,2,2-trichloroethoxycarbonyl, S-tert-butoxycarbonyl, S-benzyloxycarbonyl, S-p-methoxybenzyloxycarbonyl, S-fluorenylmethylcarbonyl, S-n-ethylaminocarbonyl, S-(N-methoxymethyl)aminocarbonyl, etc.

[0216] In a preferred embodiment, the thiol group can be protected by reacting to form a thioether, a thioester, an asymmetric disulfide bond, or a sulfinyl derivative, wherein the protecting group forming the asymmetric disulfide bond can be S-ethyl, S-tert-butyl, or a substituted S-phenyl group, etc.

[0217] In a preferred embodiment, the protection of the thiol group can be achieved by reacting to form a thioether, a thioester, an asymmetric disulfide bond, or a sulfinyl derivative. The protecting group forming the sulfinyl derivative can be: S-sulfonate derivative, S-thiosulfonate derivative, S-sulfinyl thiocarbonate, S-3-nitro-2-pyridinethio, S-[tricarbonyl[1',2,3,4,5-n]-2,4-cyclohexadien-1-yliron, oxathione isopentyl ketone, etc.

[0218] In a preferred embodiment, the dithiol group can be protected by reacting to generate a dithioacetal or ketal. The protecting group can be: S,S'-methylene, S,S'-isopropylidene, S,S'-benzylidene, or S,S'-p-methoxybenzylidene, etc.

[0219] In a preferred embodiment, the amino and thiol groups are present simultaneously, and the amino and thiol groups can be protected by reacting to generate thiazoline or ninhydrin.

[0220] In a preferred embodiment, the protecting group for the thiol group is preferably selected from the following protecting groups: diphenylmethyl (Dpm), triphenylmethyl (Trt), monomethoxytriphenylmethyl (Mmt), benzyl (Bn), p-methylbenzyl, p-methoxybenzyl (Mob), 2,4,6-trimethoxybenzyl (Tmob), 2-nitrobenzyl (oNb), 2-nitro-4,5-dimethoxybenzyl (oNv), tert-butyl (tBu), 1-adamantyl, fluorenemethyl (Fm), 2,4-dinitrophenylethyl (Dnpe), fluorenemethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), acetaminomethyl (Acm), phenylacetaminomethyl (Phacm), S-tert-butylthio (StBu), S-2-pyridinylthio (S-Pyr), S-3-nitro-2-pyridinylthio (Npys), tetrahydropyranyl (Thp), methanesulfonyl, p-toluenesulfonyl, etc.

[0221] The leaving groups mentioned above refer to the parts that leave during nucleophilic or condensation reactions, including but not limited to: H, OH, H₂O, halogens (e.g., F, Cl, Br, and I), cyanate anions, inorganic acids (e.g., nitric acid, sulfuric acid, phosphoric acid), carboxylic acids (e.g., acetic acid, trifluoroacetic acid, and benzoic acid), sulfonic acids (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, and p-nitrobenzenesulfonic acid), carbon dioxide (CO₂), nitrogen (N₂), imidazole, and alkoxy groups (RO). - ), amino (-NHR, where R is H, alkyl or aryl), phenoxy, tertiary carbocation (e.g. tert-butyl cation), carbocation stabilized by an unsaturated system or heteroatom, or any of the protecting groups described above.

[0222] The nucleophilic groups mentioned above refer to molecules or ions that can donate electron pairs to form new chemical bonds in chemical reactions. Common nucleophilic groups include: hydroxide ions (HO ions). - ), ammonia (NH3), hydroxylamine (NH2OH), hydrazine (NH2-NH2), substituted hydrazine, nucleophilic halogens (such as Cl- ... - ,Br - Or I - ), hydrogen anions (H - ), azide negative ions (N3) - ), cyanate anion (CN) - ), alcohol or alkoxy anions (e.g., alcohols with the hydroxyl hydrogen removed), amines (including primary, secondary and tertiary amines) or amine anions, carbanions (e.g., carbanions in organometallic reagents such as Grignard reagents, organolithium reagents, and Gilman reagents), thiols or thiol anions, thioethers, enols or enol anions, alkenyl ethers, enamines, carboxylic acids or carboxylic acid anions, alkyl or arylphosphine (e.g., triphenylphosphine), aromatic heterocycles with lone pairs of electrons (e.g., pyridine), etc.

[0223] In a preferred embodiment, the synthetic route for the ionizable cationic lipid compound is as follows:

[0224]

[0225] Where M' is M or M containing a protecting group.

[0226] In a preferred embodiment, the synthetic route for the ionizable cationic lipid compound is as follows:

[0227]

[0228] Where M' is M or M containing a protecting group; A is O, NH or S.

[0229] In a preferred embodiment, the synthetic route for the ionizable cationic lipid compound is as follows:

[0230]

[0231] Where X is a halogen; R4' is R4 or R4 containing a protecting group; R5' is R5 or R5 containing a protecting group.

[0232] In a preferred embodiment, the synthetic route for the ionizable cationic lipid compound is as follows:

[0233]

[0234] Where X is a halogen; R4' is R4 or R4 containing a protecting group; R5' is R5 or R5 containing a protecting group.

[0235] In a preferred embodiment, the synthetic route for the ionizable cationic lipid compound is as follows:

[0236]

[0237] Among them, M pro It is M or M containing a protecting group.

[0238] In a seventh aspect, the present invention provides the application of the aforementioned ionizable cationic lipid compounds as surfactants.

[0239] In a preferred embodiment, the ionizable cationic lipid compound is used as an emulsifier, suspending agent, dispersant, solubilizer, lubricant, thickener, antibacterial agent, or preservative.

[0240] Eighthly, the present invention provides the use of the aforementioned ionizable cationic lipid compounds in the preparation of cosmetic compositions.

[0241] In a ninth aspect, the present invention provides the use of the aforementioned ionizable cationic lipid compound in the preparation of lipid compositions, wherein the ionizable cationic lipid compound is used as a drug delivery carrier.

[0242] In a tenth aspect, the present invention provides a lipid composition, wherein the lipid component of the lipid composition comprises the aforementioned ionizable cationic lipid compound.

[0243] In a preferred embodiment, the lipid component of the lipid composition further comprises other lipids, including phospholipids, structural lipids, and PEG-conjugated lipids; the molar ratio of the ionizable cationic lipid compound to other lipid components is 1:0.2-5; the active ingredient of the lipid nanoparticle composition includes therapeutic and / or preventive agents; the therapeutic and / or preventive agents are vaccines or compounds capable of inducing an immune response; and the mass ratio of the lipid component to the active ingredient is 10:1 to 60:1.

[0244] In a preferred embodiment, the phospholipid is selected from 1,2-dilinoleoyl-sn-glycerol-3-phosphocholine.

[0245] (DLPC), 1,2-Dimyristoyl-sn-glycerol-3-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycerol-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), 1,2-distearateoyl-sn-glycerol-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycerol-3-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycerol-3-phosphocholine 1,2-Oleoyl-2-cholesterolylhemisuccino-sn-glycerol-3-phosphate choline (OChemsPC), 1-hexadecyl-sn-glycerol-3-phosphate choline (C16LysoPC), 1,2-dilinolenoyl-sn-glycerol-3-phosphate choline, 1,2-disarachidonicyl-sn-glycerol-3-phosphate choline, 1,2-bis(docosahexaenoyl)-sn-glycerol-3-phosphate choline, 1,2-dioleoyl-sn-glycerol-3-phosphate ethanolamine (DOPE), 1,2-diphydanyl-sn-glycerol-3-phosphate choline 1,2-Distearyl-sn-glycerol-3-phosphate ethanolamine (ME16.0PE), 1,2-distearyl-sn-glycerol-3-phosphate ethanolamine, 1,2-dilinoleoyl-sn-glycerol-3-phosphate ethanolamine, 1,2-dilinolenoyl-sn-glycerol-3-phosphate ethanolamine, 1,2-diarachidonicoyl-sn-glycerol-3-phosphate ethanolamine, 1,2-bis(docohexanoyl-sn-glycerol-3-phosphate ethanolamine), 1,2-dioleoyl-sn-glycerol-3-phosphate-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPP) G), at least one of palmitoyl oleoyl phosphatidyl ethanolamine (POPE), distearate-phosphatidyl-ethanolamine (DSPE), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoyl phosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-stearoyl ethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidyl ethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyl oleoyl phosphatidylcholine, lysophosphatidylcholine, and lysophosphatidyl ethanolamine (LPE).

[0246] In a preferred embodiment, the structural lipid is selected from at least one of cholesterol, coccosterol, sitosterol, ergosterol, campesterol, stigmasterol, rapeseed sterol, tomatine, ursolic acid, and α-tocopherol.

[0247] In a preferred embodiment, the PEG lipid is selected from PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

[0248] In a preferred embodiment, the therapeutic and / or preventive agent is a nucleic acid; the nucleic acid is DNA or RNA.

[0249] In a preferred embodiment, the RNA is selected from at least one of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), and messenger RNA (mRNA).

[0250] Eleventhly, the present invention provides the use of the aforementioned lipid composition in a medicament for treating diseases in mammals, the diseases being characterized by dysfunctional or abnormal protein or polypeptide activity; the diseases being selected from infectious diseases, cancer, proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases.

[0251] In a twelfth aspect, the present invention provides a pharmaceutical composition comprising the aforementioned lipid composition and a pharmaceutically acceptable carrier.

[0252] In a preferred embodiment, the pharmaceutical composition further includes a pharmaceutically active compound selected from anti-inflammatory compounds, steroids, statins, estradiol, BTK inhibitors, S1P1 agonists, glucocorticoid receptor modulators, and antihistamines.

[0253] Compared with the prior art, the present invention has the following beneficial effects:

[0254] 1. The present invention provides an ionizable cationic lipid compound having formula (1), or its salt, stereoisomer, or tautomer. The ionizable cationic lipid compound provided by the present invention is an amphiphilic lipid compound constructed by using an ionizable cationic compound or its analogue as an ionizable lipid head group, a lipid such as a fatty acid as a tail chain, and a tetrafunctional compound as a linker, and connecting the ionizable lipid head group, tail chain, and linker through biodegradable chemical bonds (such as ester bonds or amide bonds).

[0255] 2. The ionizable cationic lipid compounds (exemplary compounds) provided by this invention have a calculated c-pKa (mol network) of 5-11. Within a common pH range (0-14), the ionization state of the molecule or the ratio between different ionization states varies with the ambient pH. The c-LogP (cLogP driver) is between 5-55, exhibiting good lipophilicity. The ionizable cationic lipid compounds of this invention are pH-dependent amphiphilic lipid compounds, indicating that they can serve as excellent surfactants, particularly suitable for preparing lipid nanoparticles for use as drug delivery carriers.

[0256] 3. The ionizable cationic lipid compounds of the present invention can be used to prepare lipid compositions. The lipid compositions of mRNA prepared using the exemplary compounds have a zeta potential of -40mV to 40mV, a dispersion index of 0.01 to 0.5, a particle size of 10 to 400nm, and an encapsulation efficiency of >50%.

[0257] 4. The lipid nanoparticles of mRNA and / or other nucleic acid substances (such as siRNA, microRNA, pDNA, etc.) prepared from the ionizable cationic lipid compounds of the present invention exhibit biological activity in both in vitro cells and in vivo, such as expressing proteins encoded by the mRNA sequence. The luciferase mRNA lipid nanoparticles prepared using the exemplary compounds showed a fluorescence intensity >10 in HEK293T cells (dose: 100 ng, 96-well plate) expressing luciferase. 3 RLU; mean whole-body fluorescence signal greater than 10 after administration via tail vein injection in mice. 4 p / s / cm 2 The / sr reading indicates that the luciferase mRNA lipid nanoparticles prepared using the ionizable cationic lipid compounds of this invention exhibit biological activity in both in vitro cells and in vivo, and possess low toxicity. The lipid nanoparticles provided by this invention demonstrate good safety and efficacy as drug carriers. Attached Figure Description

[0258] Figure 1 Luciferase mRNA lipid nanoparticles prepared for compound 1005 were used to measure luciferase activity in HEK293T cells.

[0259] Figure 2 Whole-body in vivo imaging of mice 3 h and 6 h after intravenous administration of luciferase mRNA lipid nanoparticles prepared for compound 1008. Detailed Implementation

[0260] The synthetic processes of this invention are accommodating to a variety of functional groups, thus allowing the use of a wide range of substituted starting materials. These processes generally provide the desired final compound at or near the end of the process, although in some cases it may be necessary to further convert the compound into its pharmaceutically acceptable salt. The compounds of this invention can be prepared in a variety of ways using commercially available starting materials, compounds known in the literature, or from readily prepared intermediates, employing standard synthetic methods and procedures known to those skilled in the art or readily apparent to those skilled in the art based on the teachings herein. Standard synthetic methods and procedures for preparing organic molecules, as well as functional group transformations and manipulations, are available from relevant scientific literature or from standard textbooks in the field. The following description of synthetic methods is intended to illustrate, and not limit, the general procedures used to prepare the compounds of this invention.

[0261] The compounds of the present invention having the various forms described herein can be prepared from commercially available starting materials or starting materials that can be prepared using literature processes, according to the processes described in the following general synthetic routes 1, 2, 3, 4, and 5. Variables in each general synthetic route (e.g., R1, R2, and R3, etc.) are as defined herein. Those skilled in the art will note that the order of certain steps in the reaction procedures and synthetic schemes described herein can be varied, such as the introduction and removal of protecting groups.

[0262] The reaction schemes described herein can yield a variety of stereoisomers. When a specific stereoisomer is not indicated, this should be understood to include all possible stereoisomers produced by the reaction. Those skilled in the art will recognize that the reaction can be optimized to preferentially yield one isomer, or new schemes can be designed to produce a single isomer. If a mixture is produced, the isomers can be separated using techniques such as preparative thin-layer chromatography, preparative HPLC, preparative chiral HPLC, or preparative SFC.

[0263] General Synthetic Route 1

[0264]

[0265] Where M' is M or M containing a protecting group.

[0266] As described in the general synthetic route 1 above, Boc-aminotris(hydroxymethyl)methane reacts with an acid (compound 2) in a condensation reaction to generate compound 3. Step 1 can be carried out in an organic solvent (e.g., dichloromethane (DCM)) in the presence of, for example, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCl) and 4-dimethylaminopyridine (DMAP). Step 1 can be carried out at room temperature for 24 hours.

[0267] Next, compound 3 is deprotected by the Boc protecting group to generate compound 4. Step 2 can be carried out in an organic solvent (e.g., DCM) under the catalysis of an acid (e.g., trifluoroacetic acid).

[0268] Next, compound 4 and compound 5 undergo a condensation reaction to give compound 6. Step 3 can be carried out in an organic solvent (e.g., DCM or DMF) catalyzed by EDCl and DMAP or dicyclohexylcarbodiimide (DCC).

[0269] If the M' group of compound 6 contains the aforementioned protecting group, the protecting group is removed to obtain the target lipid compound. Step 4 is carried out under the selected protecting group removal reaction conditions.

[0270] General Synthesis Route 2

[0271]

[0272] Where M' is M or M containing a protecting group; A is O, NH or S.

[0273] As described in the general synthetic route 2 above, compound 1 undergoes a condensation reaction with compound 2 to give compound 3. Step 1 can be carried out in an organic solvent (e.g., DCM) in the presence of EDCl and DMAP.

[0274] Next, compound 3 is deprotected by a tert-butyloxy protecting group to give compound 4. Step 2 can be carried out in an organic solvent (e.g., DCM) in the presence of an acid (e.g., trifluoroacetic acid) and a positive ion scavenger (e.g., triisopropylsilane (TiPS)).

[0275] Next, compound 4 undergoes a condensation reaction with compound 5 to give compound 6. Step 3 can be carried out in an organic solvent (e.g., DCM or DMF) in the presence of EDCl and DMAP or DCC.

[0276] If the M' group of compound 6 contains the aforementioned protecting group, the protecting group is removed to obtain the target lipid compound. Step 4 is carried out under the deprotection reaction conditions of the selected protecting group.

[0277] General Synthesis Route 3

[0278]

[0279] Where X is a halogen, such as Cl, Br or I; R4' is R4 or R4 containing a protecting group; R5' is R5 or R5 containing a protecting group.

[0280] As described in the general synthetic route 3 above, Boc-aminotris(hydroxymethyl)methane undergoes a condensation reaction with compound 2 to generate compound 3. Step 1 can be carried out in an organic solvent (e.g., DCM) in the presence of, for example, EDCl and DMAP. Step 1 can be carried out at room temperature for 24 hours.

[0281] Next, compound 3 is deprotected by the Boc protecting group to generate compound 4. Step 2 can be carried out in an organic solvent (e.g., DCM) under the catalysis of an acid (e.g., trifluoroacetic acid).

[0282] Next, compound 4 reacts with a halogen-substituted aldehyde (compound 5) via condensation and reduction to give compound 6. Step 3 can be carried out in an organic solvent (such as dichloroethane (DCE)) in the presence of a reducing agent (e.g., sodium triacetylborohydride (NaBH(OAc)3)).

[0283] Next, compound 6 undergoes a nucleophilic reaction with an amine (compound 7) to generate compound 8. Step 4 can be carried out in an organic solvent (such as DMF), in the presence of a base (such as a non-nucleophilic organic base (e.g., triethylamine, iPr2EtN) or an inorganic base (e.g., K2CO3)) and a catalyst (KI or NaI).

[0284] If the R4' and / or R5' groups of compound 8 contain the aforementioned protecting groups, the protecting groups are removed to obtain the target lipid compound. Step 5 is carried out under the deprotection reaction conditions of the selected protecting groups.

[0285] General Synthesis Route 4

[0286]

[0287] Where X is a halogen, such as Cl, Br or I; R4' is R4 or R4 containing a protecting group; R5' is R5 or R5 containing a protecting group.

[0288] As described in the general synthetic route 4 above, Boc-aminotris(hydroxymethyl)methane undergoes a condensation reaction with compound 2 to generate compound 3. Step 1 can be carried out in an organic solvent (e.g., DCM) in the presence of, for example, EDCl and DMAP. Step 1 can be carried out at room temperature for 24 hours.

[0289] Next, compound 3 is deprotected by the Boc protecting group to generate compound 4. Step 2 can be carried out in an organic solvent (e.g., DCM) under the catalysis of an acid (e.g., trifluoroacetic acid).

[0290] Next, compound 4 undergoes a condensation reaction with a halogen-substituted compound 5 to give compound 6. Step 3 can be carried out in an organic solvent such as DCM or DMF, catalyzed by EDCl and DMAP or DCC.

[0291] Next, compound 6 undergoes a nucleophilic reaction with an amine (compound 7) to generate compound 8. Step 4 can be carried out in an organic solvent (e.g., DMF) in the presence of a base (e.g., a non-nucleophilic organic base (e.g., triethylamine, iPr₂EtN) or an inorganic base (e.g., K₂CO₃)) and a catalyst (KI or NaI).

[0292] If the R4' and / or R5' groups of compound 8 contain the aforementioned protecting groups, the protecting groups are removed to obtain the target lipid compound. Step 5 is carried out under the deprotection reaction conditions of the selected protecting groups.

[0293] General Synthesis Route 5

[0294]

[0295] Where M pro It is M or M containing a protecting group.

[0296] As described in the general synthetic route 5 above, Boc-aminotris(hydroxymethyl)methane undergoes a condensation reaction with compound 2 to generate compound 3. Step 1 can be carried out in an organic solvent (e.g., DCM) in the presence of, for example, EDCl and DMAP. Step 1 can be carried out at room temperature for 24 hours.

[0297] Next, compound 3 is deprotected by the Boc protecting group to generate compound 4. Step 2 can be carried out in an organic solvent (e.g., DCM) under the catalysis of an acid (e.g., trifluoroacetic acid).

[0298] Next, compound 4 and compound 5 undergo a condensation and reduction reaction to yield compound 6. Step 3 can be carried out in an organic solvent (such as dichloroethane (DCE)) in the presence of a reducing agent (such as sodium triacetylborohydride (NaBH(OAc)3)).

[0299] If M of compound 6 pro If the group contains the aforementioned protecting group, the protecting group is removed to obtain the target lipid compound. Step 4 is carried out under the deprotection reaction conditions of the selected protecting group.

[0300] Furthermore, it should be understood that any particular embodiment of the invention within the scope of the prior art may be expressly excluded from any one or more claims. Since such embodiments are considered to be known to those skilled in the art, they may be excluded, even if such exclusion is not expressly stated herein.

[0301] All sources cited herein, such as references, publications, databases, database entries, and techniques, are incorporated herein by reference, even if not explicitly stated in the citation. In the event of any conflict between the cited sources and the statements in this application, the statements in this application shall prevail.

[0302] Example 1: Synthesis of compounds according to formulas (1), (1A), (1B), (1C), (1D), (1E), (1F), (1G), (1H), (1I), (1J), (1K), (1L), or (1M).

[0303] A. General considerations

[0304] It is worth noting that the raw materials used in this invention are all commercially available products, and their sources are not specifically limited.

[0305] The process routes described below can be used to synthesize compounds 1001-3422 of the present invention.

[0306] The following abbreviations are used in this article:

[0307] THF: Tetrahydrofuran

[0308] MeCN: Acetonitrile

[0309] MeOH: Methanol

[0310] PE: Petroleum ether

[0311] EA: Ethyl acetate

[0312] DMF: N,N-dimethylformamide

[0313] EDCl: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride

[0314] LAH: Lithium Aluminum Hydrogen

[0315] DCM: Dichloromethane

[0316] DMAP: 4-Dimethylaminopyridine

[0317] LDA: Lithium diisopropylamino

[0318] rt: room temperature

[0319] DCE: 1,2-Dichloroethane

[0320] n-BuLi: n-Butyllithium

[0321] i-Pr2EtN:N,N-Diisopropylethylamine

[0322] B. Intermediate Synthesis

[0323] Intermediate A:

[0324]

[0325] Intermediate A is obtained using the following synthetic process:

[0326]

[0327] Tris(hydroxymethyl)aminomethane (50.0 g) and di-tert-butyl carbonate anhydride (Boc2O) (99.1 g) were dissolved in a methanol (300 mL) / H2O (30 mL) mixed solvent and reacted at room temperature for 72 h. The mixture was purified by silica gel column chromatography (DCM:MeOH = 20:1-10:1) to give a white solid intermediate A (N-Boc-aminotris(hydroxymethyl)methane) (75.0 g). LCMS (ESI) calcd for C9H 19 NO5, [M+H] + m / z 222.13, found 222.25.

[0328] Intermediate B:

[0329]

[0330] Intermediate B is obtained using the following synthetic process:

[0331]

[0332] Diethyl 2-ethyl-1,3-malonic acid (29.0 g), THF (90 mL), and DMF (30 mL) were added to a single-necked flask. NaH (3.7 g) was added under ice bath conditions, and the mixture was stirred at room temperature for 30 min under nitrogen protection. Pentadecane bromo was added, and the mixture was reacted at 80 °C for 2 h under nitrogen protection. THF was removed by concentration under reduced pressure. The reaction mixture was added dropwise to ice water, ethyl acetate was added, and the mixture was stirred. The mixture was filtered through diatomaceous earth, and the organic phase was collected and separated. The organic phase was washed with saturated brine and separated. The organic phase was purified by silica gel column chromatography (PE:EA = 100:1-50:1) to obtain compound 2-ethyl-2-pentadecanyl maleate diethyl ester (16.5 g). EtOH (50 mL), H₂O (50 mL), and KOH (11.3 g) were added to diethyl 2-ethyl-2-pentadecanyl maleate (8.0 g), and the mixture was reacted at 90 °C for 12 h. The EtOH was removed by vacuum concentration, and the reaction system was adjusted to pH 4–5 by adding dilute hydrochloric acid. Water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine and purified by column chromatography (PE:EA = 5:1–1:1) to obtain 2-ethyl-2-pentadecanyl maleic acid (5.2 g). 2-Ethyl-2-pentadecanyl maleic acid (10.0 g) was reacted at 170 °C for 6 h under open conditions, then cooled to room temperature. Water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine and separated. The organic phase was concentrated and purified by column chromatography (DCM:MeOH = 20:1–10:1) to obtain intermediate B (2-ethylheptadecanoic acid) (8.2 g). 1H NMR (400MHz, CDCl3) δ2.31 (tt, J = 8.6, 5.3Hz, 1H), 1.71–1.46 (m, 4H), 1.28 (s, 26H), 0.96 (t, J = 7.4Hz, 3H), 0.90 (t, J = 6.7Hz, 3H).

[0333] Intermediate C:

[0334]

[0335] Intermediate C is obtained using the following synthetic process:

[0336]

[0337] Decanoic acid (50 g) and solvent THF (500 mL) were added to a three-necked flask. After cooling the system to 0 °C, reactant NaH (23.22 g) was slowly added, and the mixture was stirred for 1 h under nitrogen protection at 0 °C. Then, LDA (62.19 g) was slowly added dropwise, and the mixture was stirred for another 1 h under nitrogen protection at 0 °C. Finally, iodononane (88.52 g) was added dropwise, and the mixture was heated to room temperature and stirred overnight. The mixture was then diluted with 1 LDCM and washed with saturated NH4Cl solution and water, respectively. The organic layer was dried over anhydrous Na2SO4, filtered, concentrated, and purified by silica gel chromatography (PE:EA = 50:1) to obtain intermediate C (2-octylundecanoic acid) (11 g). 1HNMR(400MHz,Chloroform-d)δ2.37(tt,J=8.7,5.1Hz,1H),1.71–1.58(m,2H ), 1.48 (dt, J = 13.3, 6.7Hz, 2H), 1.29 (d, J = 9.4Hz, 25H), 0.90 (t, J = 6.8Hz, 6H).

[0338] Intermediate D:

[0339]

[0340] Intermediate D is obtained using the following synthetic process:

[0341]

[0342] Add 79.5 g of tridecanoic acid and 800 mL of THF solvent to a three-necked flask. After cooling the system to 0 °C, slowly add NaH (22.25 g). Stir the reaction solution at 0 °C under nitrogen protection for 1 h. Then, slowly add LDA (317.87 g) dropwise to the reaction system. Continue stirring at 0 °C under nitrogen protection for another 1 h. After adding n-hexane iodide (94.3 g) dropwise, raise the temperature to room temperature and stir overnight. Dilute the reaction solution with 1 LDCM and wash with saturated NH4Cl solution and water, respectively. Dry the organic layer with anhydrous Na2SO4, filter, concentrate, and perform silica gel column chromatography (PE:EA = 50:1) to obtain intermediate D (2-hexyltridecanoic acid) (26 g). 1H NMR(400MHz,Chloroform-d)δ2.36(tt,J=8.7,5.4Hz,1H),1.63(ddd,J=14.3,8.7,5.5 Hz, 2H), 1.49 (dq, J = 13.5, 6.6 Hz, 2H), 1.29 (d, J = 10.1 Hz, 26H), 0.90 (t, J = 6.6 Hz, 6H).

[0343] Intermediate E:

[0344]

[0345] Intermediate E is obtained using the following synthetic process:

[0346]

[0347] Compound tris(hydroxymethyl)aminomethane (10.0 g) and tert-butyl acrylate (21.1 g) were dissolved in EtOH (150 mL). Under nitrogen protection, the reaction was carried out at 45 °C for 30 h. After removing EtOH by vacuum concentration, 100 mL × 3 of solvent (PE:EA = 20:1) was added and the mixture was stirred and filtered to obtain a white solid intermediate compound E (11.5 g). ¹H NMR (400 MHz, CDCl₃) δ 3.60 (s, 6 H), 2.84 (t, J = 5.9 Hz, 2 H), 2.47 (t, J = 5.8 Hz, 2 H), 1.48 (s, 9 H).

[0348] Intermediate F:

[0349]

[0350] Intermediate F was obtained using the following synthetic process:

[0351]

[0352] To a single-necked flask, 2.7 g of (2-aminoethyl)carbamate tert-butyl ester, 90 mL of MeCN, 7.99 g of benzyl 2-bromoethyl ether, and 11.65 g of K₂CO₃ were added. The mixture was reacted overnight at 80 °C. Water and ethyl acetate were then added, and the mixture was stirred and separated. The organic phase was washed with saturated brine. The organic phase was separated, concentrated, and purified by silica gel column chromatography (PE:EA = 10:1-5:1) to obtain compound 2 (5.5 g). Dioxane (30 mL) and dioxane hydrochloride solution (30 mL) were added to compound 2. After stirring at room temperature for 3 h, the mixture was concentrated under reduced pressure to obtain compound 3, i.e., intermediate F (6 g). LCMS(ESI)calcd for C 20 H 28 N₂O₂, [M+H] + m / z329.22, found 329.24.

[0353] Intermediate G:

[0354]

[0355] Intermediate G is obtained using the following synthetic process:

[0356]

[0357] Compound 1 (3.0 g), MeCN (90 mL), 3-benzyloxybromopropane (9.45 g), and K₂CO₃ (12.94 g) were added to a single-necked flask. The mixture was reacted overnight at 80 °C. Water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, and the organic phase was separated, concentrated, and purified by column chromatography (PE:EA = 10:1-5:1) to obtain compound 2 (6.9 g). Dioxane (30 mL) and dioxane hydrochloride solution (30 mL) were added to compound 2 (6.9 g). The mixture was stirred at room temperature for 3 h, and then concentrated under reduced pressure to obtain intermediate G (7.5 g). LCMS(ESI)calcd for C 22 H 32 N₂O₂, [M+H] + m / z 357.25, found 357.51.

[0358] Intermediate H:

[0359]

[0360] Intermediate H was obtained using the following synthetic process:

[0361]

[0362] Compound 1 (1.5 g), MeCN (90 mL), 4-bromobutyl ether benzyl ester (5.02 g), and K₂CO₃ (6.47 g) were added to a single-necked flask. After reacting overnight at 80 °C, water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, and the organic phase was separated, concentrated, and purified by column chromatography (PE:EA = 20:1-10:1) to obtain compound 2 (4.1 g). Dioxane (30 mL) and dioxane hydrochloride solution (30 mL) were added to compound 2 (4.1 g). After stirring at room temperature for 3 h, the mixture was concentrated under reduced pressure to obtain intermediate H (4.9 g). LCMS (ESI) calcd for C 24 H 36 N₂O₂, [M+H] + m / z 385.28, found 385.56.

[0363] Intermediate I:

[0364]

[0365] Intermediate I was obtained using the following synthetic process:

[0366]

[0367] Compound 1 (3.9 g), MeCN (90 mL), benzyl 2-bromoethyl ether (5.78 g), and K₂CO₃ (15.46 g) were added to a single-necked flask. The mixture was reacted overnight at 80 °C. Water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, concentrated, and purified by column chromatography (PE:EA = 20:1-10:1) to obtain compound 2 (6.8 g). Dioxane (30 mL) and dioxane hydrochloride solution (30 mL) were added to compound 2 (6.8 g). The mixture was stirred at room temperature for 3 h, then concentrated under reduced pressure to obtain intermediate I (6.5 g). LCMS(ESI)calcd for C 12 H 20 N₂O, [M+H] + m / z 209.16, found 209.31.

[0368] Intermediate J:

[0369]

[0370] Intermediate J is obtained using the following synthetic process:

[0371]

[0372] Butyric acid (5.0 g) and THF (100 mL) were added to a single-necked flask. NaH (2.73 g) was added at 0 °C, followed by slow dropwise addition of LDA (56.8 mL). The reaction was carried out at room temperature for 30 min. 1-Bromotridecane was added, and the reaction continued overnight at room temperature. Ice water and ethyl acetate were then added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, concentrated, and purified by column chromatography (PE:EA = 10:1-5:1) to obtain intermediate J (5.0 g). LCMS(ESI)calcd for C 17 H 34 O2, [M+H] + m / z271.26, found 271.46.

[0373] Intermediate K:

[0374]

[0375] Intermediate K was obtained using the following synthetic process:

[0376]

[0377] Reactant 1 (24 g), imidazole (19.00 g), and solvent DCM (200 mL) were added to a three-necked flask. After cooling to 0 °C, reactant TBDMSCI (38.57 g) was slowly added. The mixture was stirred at 25 °C for 4 h, then diluted with 300 mL of DCM. The organic phase was washed twice with 1 L of water and dried over anhydrous Na2SO4. After filtration and concentration, the mixture was filtered through a 1:20 DCM:MeOH column and collected. The sample was concentrated to obtain intermediate K (28.00 g). ¹H NMR (400 MHz, CDCl3) δ 4.89 (s, 1H), 3.58 (t, J = 6.1 Hz, 2H), 2.66 (t, J = 7.3 Hz, 2H), 2.45 (s, 3H), 1.67–1.40 (m, 4H), 0.84 (s, 9H).

[0378] Intermediate L:

[0379]

[0380] Intermediate L was obtained using the following synthetic process:

[0381]

[0382] Compound DMSO (3.58 g) and anhydrous solvent DCM (30 mL) were added to a three-necked flask. The system was cooled to -78 °C, and oxaloyl chloride (2.91 g) was slowly added. The reaction mixture was stirred at -78 °C under N2 protection for 10 minutes. Then, compound 1 (3 g) was slowly added dropwise. The mixture was stirred at -78 °C under N2 protection for 1 hour. Finally, TEA (9.28 g) was added dropwise, and the mixture was stirred at -78 °C under N2 protection for 0.5 hours. The mixture was then diluted with 100 mL of DCM and washed with saturated NH4Cl solution and water, respectively. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to obtain intermediate L (3.0 g). LCMS (ESI) calcd for C 11 H 14 O3, [M+H] + m / z195.09, found195.23.

[0383] Intermediate M:

[0384]

[0385] Intermediate M is obtained using the following synthetic process:

[0386]

[0387] Intermediate K (2.0 g), compound 1 (1.79 g), and solvent ACN (20 mL) were added to a sealed tube. Reactants K₂CO₃ (3.81 g) and KI (1.52 g) were then added. The mixture was stirred overnight at 70 °C under N₂ protection. The solution was diluted with 100 mL of EA and washed with saturated NH₄Cl solution and water, respectively. The organic layer was dried over anhydrous Na₂SO₄, filtered, concentrated, and then filtered through a 15:1 DCM:MeOH column in silica gel. The sample was collected and concentrated to obtain compound 2 (2.0 g). Reactant 2 (1.80 g) and solvent DCM (20 mL) were added to a three-necked flask. After purging the system with nitrogen, the temperature was lowered to -78 °C. Then, DIBAL-H (1.00 g) was added dropwise, and the mixture was stirred at -78 °C for 4 h. The reaction mixture was then quenched by adding methanol and sodium carbonate solution dropwise. Extraction was performed with DCM (100 mL), and the organic layer was dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to obtain intermediate M (1.36 g). LCMS(ESI)calcdforC 15 H 33 NO2Si,[M+H] + m / z288.52,found288.23.

[0388] C. Compound 1003 was synthesized according to general synthetic route 1.

[0389] Structural formula:

[0390] Chemical formula: C 63 H 122 N2O7

[0391] Molecular weight: 1019.68

[0392] Step 1: Synthesize compound 3 in general synthetic route 1

[0393]

[0394] Compound 1 (intermediate A) (1.0 g), DCM (20 mL), DMAP (2.2 g), and EDCI (3.4 g) were added to compound 2 (intermediate B) (4.3 g). The mixture was stirred at room temperature for 12 h under nitrogen protection. Water and dichloromethane were then added and the mixture was stirred and separated. The organic phase was washed with saturated brine to separate the organic phase. The organic phase was then dried and concentrated with anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 40:1-30:1) to obtain compound 3 (2.6 g).

[0395] Step 2: Synthesize compound 4 from general synthetic route 1

[0396]

[0397] DCM (15 mL) and TFA (5 mL) were added to compound 3 (2.6 g), stirred at room temperature for 3 h, concentrated, and purified by silica gel column chromatography (PE:EA = 10:1-5:1) to obtain compound 4 (3.2 g).

[0398] Step 3: Synthesize compound 6 from general synthetic route 1

[0399]

[0400] DCM (20 mL), Boc-glycine (compound 5, 578 mg), and DCC (6.8 g) were added to compound 4 (3.2 g). The mixture was stirred at room temperature under nitrogen protection for 12 h. Water and ethyl acetate were then added and the mixture was stirred and separated. The organic phase was washed with saturated brine and separated. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE:EA = 5:1-2:1) to obtain compound 6 (2.2 g).

[0401] Step 4: Compound 1003

[0402]

[0403] Add DCM (15 mL) and TFA (5 mL) to compound 6 (2.2 g), stir at room temperature for 12 h, concentrate, and purify by column chromatography (DCM:MeOH = 30:1-20:1) to obtain compound 1003 (1.3 g). 1 H NMR (400MHz, CDCl3) δ4.49 (d, J=2.6Hz, 6H), 3.28 (s, 2H), 2.32 (tt, J=8.6, 5. 5Hz, 3H), 1.62 (q, J = 7.1Hz, 12H), 1.27 (d, J = 3.4Hz, 78H), 0.94-0.86 (m, 18H).

[0404] D. Compound 1002, synthesized according to general synthetic route 1.

[0405] Structural formula:

[0406] Chemical formula: C 60 H 116 N2O7

[0407] Molecular weight: 977.60

[0408] The synthesis was carried out according to general synthetic route 1, which is similar to the synthesis process of compound 1003. The difference is that intermediate C was used as compound 2 in general synthetic route 1. 1H NMR (400MHz, CDCl3) δ4.48 (s, 6H), 3.27 (s, 2H), 2.37 (tt, J = 8.5, 5.5Hz, 3H), 1 .64(d,J=6.9Hz,6H),1.50-1.42(m,6H),1.27(s,72H),0.90(t,J=6.7Hz,18H).

[0409] Compound E.1004, synthesized according to general synthetic route 1.

[0410] Structural formula:

[0411] Chemical formula: C 54 H 104 N2O7

[0412] Molecular weight: 893.43

[0413] The synthesis was carried out according to general synthetic route 1, which is similar to the synthesis process of compound 1003, except that palmitic acid was used as compound 2 in general synthetic route 1. 1 H NMR (400MHz, CDCl3) δ7.55 (s, 1H), 4.45 (s, 6H), 3.74 (s, 2H), 2.35 (t, J = 7.6Hz, 6H), 1.60 (p, J = 6.9Hz, 6H), 1.27 (s, 72H), 0.90 (t, J = 6.7Hz, 9H).

[0414] Compound F. 1001, synthesized according to general synthetic route 1.

[0415] Structural formula:

[0416] Chemical formula: C 59 H 108 N2O7

[0417] Molecular weight: 957.52

[0418] The synthesis was carried out according to general synthetic route 1, which is similar to the synthesis process of compound 1003. The difference is that (9Z)-9-hexadecenoic acid is used as compound 2 in general synthetic route 1, and 5-(N,N-dimethylamino)valerate is used as compound 5 in general synthetic route 1. 1H NMR (400MHz, CDCl3) δ5.42-5.30(m,6H),4.51-4.38(m,6H),3.19(dt,J=11.3,5 .6Hz,2H),2.97(t,J=5.9Hz,6H),2.46(t,J=6.6Hz,1H),2.35(td,J=7.6,3.1Hz, 5H),2.20(t,J=7.6Hz,2H),2.03(d,J=6.2Hz,12H),1.89-1.81(m,3H),1.74(q, J=7.8Hz,2H),1.67-1.51(m,6H),1.31(d,J=7.8Hz,48H),0.90(t,J=6.7Hz,9H).

[0419] Compound G.1014, synthesized according to general synthetic route 2.

[0420] Structural formula:

[0421] Chemical formula: C 70 H 137 N3O9

[0422] Molecular weight: 1164.88

[0423] Step 1: Synthesize the general synthetic route 2 intermediate 3

[0424]

[0425] Intermediate 3 was synthesized according to step 1 of general synthetic route 1, except that intermediate E was used as compound 1 in general synthetic route 1.

[0426] Step 2: Synthesize compound 4 of general synthetic route 2

[0427]

[0428] DCM (15 mL), TFA (12 mL), and TiPS (3 mL) were added to compound 3 (4.8 g). After reacting at room temperature for 12 h, water and dichloromethane were added and the mixture was stirred and separated. The organic phase was washed with saturated brine, and the organic phase was separated, concentrated, and purified by silica gel column chromatography (DCM:MeOH = 20:1-10:1) to obtain compound 4 (3.3 g).

[0429] Step 3: Synthesize compound 6 of general synthetic route 2

[0430]

[0431] Compound 4 (4.0 g), DCM (80 mL), EDCI (1.1 g), 1-hydroxybenzotriazole (HOBt) (0.78 g), and DIEA (2.5 g) were added to compound 5 (1.9 g). After stirring at room temperature for 12 h, water and dichloromethane were added and the mixture was stirred and separated. The organic phase was washed with saturated brine, and the organic phase was separated, concentrated, and purified by silica gel column chromatography (DCM:MeOH = 30:1-20:1) to obtain compound 6 (4.0 g).

[0432] Step 4: Compound 1014

[0433]

[0434] To compound 6 (4.0 g), MeOH (30 mL), DCM (10 mL), and Pd / C (4.73 g (10%)) were added. The mixture was stirred overnight at room temperature under hydrogen atmosphere. The mixture was then filtered through diatomaceous earth, concentrated, and subjected to silica gel column chromatography (DCM:MeOH 30:1).

[0435] 15:1) yielded compound 1014 (2g). 1 H(400MHz, CDCl3)δ7.38(t,J=5.8Hz,1H),4.14-4.05(m,6H),3.58(t,J=4.9Hz,4H),3.31(q,J=5.7Hz,2H),2.89(t,J=6.3Hz, 2H), 2.64 (q, J=5.2Hz, 6H), 2.29 (dq, J=10.8, 4.4Hz, 6H), 1.66-1.37 (m, 12H), 1.24 (d, J=4.3Hz, 78H), 0.87 (t, J=7.1Hz, 18H).

[0436] Compound H.1015 was synthesized according to general synthetic route 2.

[0437] Structural formula:

[0438] Chemical formula: C 72 H 141 N3O9

[0439] Molecular weight: 1192.93

[0440] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that intermediate G was used as compound 5 in general synthetic route 2. 1H(400MHz, CDCl3)δ7.58(t,J=5.7Hz,1H),4.15-4.01(m,6H),3.70(t,J=5.4Hz,4H),3.39(q,J=5.7Hz,2H),2.86(t,J=6.2Hz,2H),2 .57(dt,J=20.5,5.8Hz,6H),2.36-2.22(m,5H),1.76-1.67(m,4H),1.62-1.35(m,13H),1.23(d,J=4.6Hz,78H),0.92-0.80(m,18H).

[0441] I. Compound 1020, synthesized according to general synthetic route 2.

[0442] Structural formula:

[0443] Chemical formula: C 74 H 145 N3O9

[0444] Molecular weight: 1220.99

[0445] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that intermediate H was used as compound 5 in general synthetic route 2.

[0446] Compound J.1025, synthesized according to general synthetic route 2.

[0447] Structural formula:

[0448] Chemical formula: C 69 H 135 N3O8

[0449] Molecular weight: 1134.85

[0450] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that intermediate I was used as compound 5 in general synthetic route 2. 1 H(400MHz, CDCl3)δ6.98(t,J=5.7Hz,1H),4.10(m,J=3.8Hz,6H),3.63(t,J=5.2Hz,2H),3.35(q,J=5.8Hz,2H),2.89(t,J=6.1Hz,2 H), 2.57 (q, J = 5.3Hz, 4H), 2.32 (s, 3H), 2.28 (q, J = 4.1Hz, 5H), 1.65-1.41 (m, 13H), 1.24 (d, J = 4.4Hz, 78H), 0.87 (t, J = 7.1Hz, 18H).

[0451] Compound K, 1005, was synthesized according to general synthetic route 2.

[0452] Structural formula:

[0453] Chemical formula: C 68 H 133 N3O7

[0454] Molecular weight: 1104.83

[0455] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that N,N-dimethylethylenediamine was used as compound 5 in general synthetic route 2. 1 H NMR (400MHz, CDCl3) δ4.19-4.07(m,6H),3.40(d,J=5.7Hz,2H),2.95-2.87(m,2H),2.82(s,4H),2.64-2.55(m,2H),2.38 (s, 6H), 2.33-2.29 (m, 3H), 1.63-1.55 (m, 6H), 1.49-1.44 (m, 2H), 1.26 (d, J = 5.4Hz, 78H), 0.89 (td, J = 7.0, 2.8Hz, 18H).

[0456] Compound L. 1006 was synthesized according to general synthetic route 2.

[0457] Structural formula:

[0458] Chemical formula: C 53 H 103 N3O7

[0459] Molecular weight: 894.42

[0460] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014. The difference is that tetradecanoic acid is used as compound 2 in general synthetic route 2 and N,N-dimethylethylenediamine is used as compound 5 in general synthetic route 2. 1 HNMR (400MHz, CDCl3) δ7.99 (t, J=5.8Hz, 1H), 4.13 (s, 6H), 3.60 (q, J=5.3

[0461] Hz,2H),3.38-3.34(m,2H),3.00(s,6H),2.94(t,J=5.8Hz,2H),2.45(t,J=5.8Hz,2H) ,2.35(t,J=7.6Hz,6H),1.66-1.56(m,6H),1.33-1.23(m,60H),0.90(t,J=6.8Hz,9H).

[0462] Compound M.1007 was synthesized according to general synthetic route 2.

[0463] Structural formula:

[0464] Chemical formula: C 68 H 133 N3O7

[0465] Molecular weight: 1104.83

[0466] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014. The difference is that intermediate D is used as compound 2 in general synthetic route 2 and N,N-dimethylethylenediamine is used as compound 5 in general synthetic route 2. 1 HNMR(400MHz,Chloroform-d)δ7.13(t,J=5.0Hz,1H),4.13(s,6H),3.33(q,J=5.6Hz,2H),2.92(t,J=6.0Hz,2H),2.47-2.28(m,7H ), 2.24 (s, 6H), 1.60 (dq, J = 14.7, 7.2Hz, 6H), 1.47 (dd, J = 14.2, 6.7Hz, 6H), 1.27 (d, J = 3.4Hz, 78H), 0.89 (td, J = 6.7, 2.5Hz, 18H).

[0467] Compound N.1008, synthesized according to general synthetic route 2.

[0468] Structural formula:

[0469] Chemical formula: C 68 H 133 N3O7

[0470] Molecular weight: 1104.01

[0471] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014. The difference is that intermediate C is used as compound 2 in general synthetic route 2 and N,N-dimethylethylenediamine is used as compound 5 in general synthetic route 2. 1HNMR(400MHz,Chloroform-d)δ4.13(s,6H),3.33(q,J=5.6Hz,2H),2.92(t,J=5.9Hz,2H),2.51-2.26(m,7H),2.24 (s, 6H), 1.59 (ddt, J = 14.8, 10.9, 6.4Hz, 6H), 1.47 (tq, J = 11.0, 5.4Hz, 6H), 1.26 (s, 77H), 0.89 (t, J = 6.7Hz, 18H).

[0472] Compound 1009 was synthesized according to general synthetic route 2.

[0473] Structural formula:

[0474] Chemical formula: C 62 H 121 N3O7

[0475] Molecular weight: 1020.66

[0476] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014. The difference is that intermediate J is used as compound 2 in general synthetic route 2 and N,N-dimethylethylenediamine is used as compound 5 in general synthetic route 2. 1 H(400MHz, CDCl3)δ7.04(t,J=5.0Hz,1H),4.12(m,J=3.3Hz,6H),3.30(q,J=5.6Hz,2H),2.91(t,J=6.0Hz,2H),2.39 (t,J=6.0Hz,2H),2.33-2.25(m,5H),2.22(s,6H),1.76-1.54(m,12H),1.24(m,J=3.7Hz,66H),0.90-0.84(m,18H).

[0477] Compound P.1011 was synthesized according to general synthetic route 2.

[0478] Structural formula:

[0479] Chemical formula: C 70 H 137 N3O7

[0480] Molecular weight: 1132.88

[0481] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that N,N-diethylethylenediamine was used as compound 5 in general synthetic route 2. 1H NMR (400MHz, CDCl3) δ4.19-4.08(m,6H),3.31(q,J=5.8Hz,2H),2.92(t,2H),2.57(t,J=6.9Hz,6H),2.37-2.26( m,5H),1.62(ddd,J=18.3,9.0,4.3Hz,12H),1.27(d,J=4.7Hz,78H),1.04(t,J=7.1Hz,6H),0.94-0.86(m,18H).

[0482] Q. Synthesize compound 1012 according to general synthetic route 2.

[0483] Structural formula:

[0484] Chemical formula: C 72 H 141 N3O7

[0485] Molecular weight: 1160.93

[0486] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that N,N-diethylethylenediamine was used as compound 5 in general synthetic route 2. 1 H NMR (400MHz, CDCl3) δ6.59 (s, 1H), 4.18-4.07 (m, 6H), 3.29 (q, J = 5.7Hz, 2H), 2.91 (t, J = 6.2Hz, 2H), 2.54 (t, J = 6.1 Hz, 2H), 2.40 (t, J = 7.5Hz, 4H), 2.35-2.27 (m, 5H), 1.63-1.42 (m, 16H), 1.27 (m, J = 4.5Hz, 78H), 0.93-0.85 (m, 24H).

[0487] Compound R, 1013, was synthesized according to general synthetic route 2.

[0488] Structural formula:

[0489] Chemical formula: C 74 H 145 N3O7

[0490] Molecular weight: 1188.99

[0491] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that N,N-diethylethylenediamine was used as compound 5 in general synthetic route 2. 1H NMR (400MHz, CDCl3) δ4.10(d,J=4.1Hz,6H),3.26(q,J=5.8Hz,2H),2.89(t,J=6.2Hz,2H),2.52(d,J=6.6Hz,2H) ,2.41(t,J=7.4Hz,4H),2.33-2.25(m,5H),1.60(d,J=11.8Hz,16H),1.24(d,J=4.5Hz,82H),0.96-0.79(m,24H).

[0492] Compound S. 1029, synthesized according to general synthetic route 2.

[0493] Structural formula:

[0494] Chemical formula: C 68 H 132 N2O8

[0495] Molecular weight: 1105.81

[0496] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014, except that N,N-diethyl-2-hydroxyethylamine was used as compound 5 in general synthetic route 2. 1 H NMR (400MHz, CDCl3) δ4.16(t,J=5.8Hz,2H),4.13-4.05(m,6H),2.86(t,J=6.3Hz,2H),2.55(t,J=5.8Hz,2H) ,2.44(t,J=6.3Hz,2H),2.35-2.20(m,9H),1.60-1.39(m,11H),1.24(d,J=4.5Hz,80H),1.02-0.79(m,18H).

[0497] Compound T.1111 was synthesized according to general synthetic route 2.

[0498] Structural formula:

[0499] Chemical formula: C 70 H 136 N4O7

[0500] Molecular weight: 1145.88

[0501] Synthesized according to general synthetic route 2, similar to the synthesis of compound 1014, the difference being that intermediate C was used as compound 2 in general synthetic route 2 and 4-(2-aminoethyl)piperazine-1-carboxylic acid tert-butyl ester was used as compound 5 in general synthetic route 2. ¹H NMR (400 MHz, Chloroform-d) δ 4.28 (s, 6H), 3.41 (s, 2H), 3.31 (s, 4H), 3.22 (s, 3H), 2.97–2.80 (m, 5H), 2.70 (s, 5H), 2.46–2.32 (m, 4H), 1.60 (t, J = 7.1 Hz, 6H), 1.54–1.40 (m, 6H), 1.27 (s, 78H), 0.89 (t, J = 6.7 Hz, 18H).

[0502] Compound U.1118 was synthesized according to general synthetic route 2.

[0503] Structural formula:

[0504] Chemical formula: C 71 H 139 N3O8

[0505] Molecular weight: 1162.91

[0506] The synthesis was carried out according to general synthetic route 2, which is similar to the synthesis process of compound 1014. The difference is that intermediate C is used as compound 2 in general synthetic route 2 and intermediate K is used as compound 5 in general synthetic route 2. 1 HNMR(400MHz,Chloroform-d)δ4.11(s,6H),3.61(s,2H),3.42(d,J=33.5Hz,2H),2.91(t,J=6.2Hz,3 H), 2.36 (dd, J = 9.2, 3.6Hz, 6H), 1.47 (dd, J = 13.9, 6.2Hz, 14H), 1.27 (s, 71H), 0.90 (t, J = 6.7Hz, 18H).

[0507] Compound V.1010, synthesized according to general synthetic route 3.

[0508] Structural formula:

[0509] Chemical formula: C 67 H 133 N3O6

[0510] Molecular weight: 1076.82

[0511] Steps 1 and 2: Synthesize compounds 3 and 4 of general synthetic route 3.

[0512]

[0513] The steps 1 and 2 of the general synthetic route 1 are the same as those for the synthesis of compound 1003.

[0514] Step 3: Synthesize compound 6 of general synthetic route 3

[0515]

[0516] Compound 4 (600 mg), chloroacetaldehyde (compound 5, 242 mg (40%)), and NaBH(OAc)3 (394 mg) were added to DCE (60 mL). After reacting overnight at room temperature, water and dichloromethane were added, and the mixture was stirred and separated. The organic phase was washed with saturated brine, and the organic phase was separated, concentrated, and subjected to silica gel column chromatography (DCM:MeOH = 30:1-15:1) to obtain compound 6 (270 mg).

[0517] Step 4: Synthesize compound 1010

[0518]

[0519] To compound 6 (270 mg), MeCN (20 mL), N,N-dimethylethylenediamine (compound 7, 229 mg), KI (43 mg), and K2CO3 (179 mg) were added. After reacting overnight at 70 °C under nitrogen protection, compound 1010 (16 mg) was obtained by silica gel column chromatography. 1 H(400MHz, CDCl3)δ4.10(m,J=2.9Hz,6H),2.76-2.64(m,6H),2.40(t,J=6.2Hz,2H),2.29(ddd,J=8.5,5.6 ,2.9Hz,3H),2.22(s,6H),1.97(s,6H),1.50-1.41(m,6H),1.24(d,J=4.2Hz,78H),0.87(t,J=7.1Hz,18H).

[0520] Compound W.1059 was synthesized according to general synthetic route 3.

[0521] Structural formula:

[0522] Chemical formula: C 67 H 132 N2O7

[0523] Molecular weight: 1077.80

[0524] The synthesis follows general synthetic route 3, similar to the synthesis of compound 1010, except that intermediates C and L are used as compounds 2 and 5 in general synthetic route 3, respectively. Furthermore, compound 6 in general synthetic route 3 requires functional group transformation through the following process steps:

[0525]

[0526] To intermediate 6a (1.5 g), solvents MeOH (5 mL) and THF (5 mL) were added. Reactants Pd(OH)₂ / C (10%) (0.46 g) and Pd / C (10%) (0.35 g) were added. The mixture was reacted overnight at 25 °C under H₂ conditions. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated and subjected to silica gel column chromatography (PE:EA 30:1–10:1) to obtain intermediate 6b (520 mg). To intermediate 6b (470 mg), DCM (5 mL) was added, and SOCl₂ (1064.26 mg, 8.946 mmol) was added dropwise at 0 °C. After reacting at room temperature for 3 hours, the reaction mixture was extracted with water and DCM. The organic phase was concentrated and subjected to silica gel column chromatography (PE:EA 20:1) to obtain intermediate 6c (181 mg). Dimethylamine (143.39 mg) and DMF (2 mL), potassium carbonate (43.96 mg) and potassium iodide (26.39 mg) were added to intermediate 6c (170 mg). The reaction mixture was stirred at 70 °C, filtered, concentrated, and subjected to silica gel column chromatography (DCM:MeOH20:1) to give compound 1059 (91 mg, yield 53.22%). 1 H NMR(400MHz,Chloroform-d)δ4.11(s,6H),3.51(dt,J=18.3,5.6Hz,4H),2.81(dd,J=10.3,5.1Hz,2H),2.50(t,J=6.0 Hz, 2H), 2.42-2.32 (m, 3H), 2.28 (s, 6H), 2.08-1.98 (m, 1H), 1.53-1.38 (m, 7H), 1.27 (s, 77H), 0.90 (t, J = 6.7Hz, 18H).

[0527] Compound X.1112 was synthesized according to general synthetic route 3.

[0528] Structural formula:

[0529] Chemical formula: C 69 H 136 N2O6

[0530] Molecular weight: 1089.85

[0531] The synthesis was carried out according to general synthetic route 3, which is similar to the synthesis process of compound 1010. The difference is that intermediate C and 6-bromohexanol are used respectively in general synthetic route 3 for compounds 2 and 5. 1 H NMR(400MHz,Chloroform-d)δ4.11(s,6H),3.11(s,1H),2.57(t,J=6.8Hz,2H),2.44-2.19 (m,12H),2.04(q,J=6.5Hz,2H),1.53-1.38(m,13H),1.27(s,92H),0.90(t,J=6.7Hz,20H).

[0532] Compound Y.1113 was synthesized according to general synthetic route 4.

[0533] Structural formula:

[0534] Chemical formula: C 71 H 138 N2O8

[0535] Molecular weight: 1147.89

[0536] Steps 1 and 2: Synthesize compounds 3 and 4 of general synthetic route 4.

[0537]

[0538] The steps 1 and 2 of the general synthetic route 1 are the same as those for the synthesis of compound 1003.

[0539] Step 3: Synthesize compound 6 of general synthetic route 4

[0540]

[0541] DIEA (0.30 g) and solvent DCM (20 mL) were added to compound 4. Under nitrogen protection, the mixture was cooled to 0 °C, and then 5-bromopentanoyl chloride (compound 5, 0.27 g) was added. The mixture was stirred at 25 °C for 2 h. After dilution with 100 mL of DCM, the organic phase was washed twice with 100 mL of water. The organic phase was dried over anhydrous Na2SO4, filtered, concentrated, and then subjected to silica gel column chromatography (PE:EA20:1) to give compound 6 (800 mg).

[0542] Step 4: Synthesize compound 8 of general synthetic route 4

[0543]

[0544] Compound 6 (750.00 mg), intermediate K (compound 7, 217.31 mg), and solvent DMF (20 mL) were added to a three-necked flask. Reactants K₂CO₃ (184.16 mg) and KI (110.60 mg) were then added. The mixture was stirred at 25 °C for 16 h, followed by stirring at 100 °C for 3 h. The reaction solution was diluted with 200 mL of EA and then washed twice with 300 mL of saturated brine. The organic phase was dried over anhydrous Na₂SO₄, filtered, concentrated, and then filtered through a 20:1 DCM:MeOH column after adding silica gel. The collected sample was concentrated to obtain compound 8 (700.00 mg).

[0545] Step 5: Synthesize compound 1113

[0546]

[0547] Compound 8 (700.00 mg) and solvent THF (5 mL) were added to a three-necked flask, followed by 5.55 mL of 1 M dioxane hydrochloride solution. The reaction mixture was stirred at 25 °C for 2 h, diluted with 50 mL of EA, and then washed with saturated NaHCO3 solution and water. The organic phase was dried over anhydrous Na2SO4, filtered, concentrated, and then subjected to silica gel column chromatography (MeOH:DCM1:20) to give compound 1113 (201.1 mg, yield 31.59%). 1 H NMR (400MHz, CDCl3) δ4.50-4.39(m,6H),3.59(t,J=4.6Hz,2H),2.49(s,3H),2.35-2.26(m,6H),2.14 (t,J=7.5Hz,2H),1.75-1.69(m,4H),1.66-1.43(m,17H),1.27(d,J=3.7Hz,78H),0.94-0.87(m,18H).

[0548] Compound Z.1115 was synthesized according to general synthetic route 4.

[0549] Structural formula:

[0550] Chemical formula: C 67 H 131 N3O7

[0551] Molecular weight: 1090.80

[0552] The synthesis was carried out according to general synthetic route 4, which is similar to the synthesis process of compound 1010. The difference is that 2-bromoacetyl chloride and N,N-dimethylethylenediamine were used to synthesize compounds 5 and 7 in general synthetic route 4, respectively. 1 H NMR (400)

[0553] MHz, CDCl3) δ7.42(s,1H),4.48(tt,J=11.9,5.7Hz,6H),3.27(s,2H),3.13(s,2H),3.01(s,2H),2.83(s,6H),2 .38-2.25(m,3H),1.55(dtt,J=36.3,14.1,7.3Hz,12H),1.27(d,J=3.3Hz,78H),0.90(td,J=7.2,3.4Hz,18H).

[0554] Compound 1114, synthesized according to general synthetic route 5.

[0555] Structural formula:

[0556] Chemical formula: C 70 H 138 N2O7

[0557] Molecular weight: 1119.88

[0558] Steps 1 and 2: Synthesize compounds 3 and 4 of general synthetic route 5.

[0559]

[0560] The steps 1 and 2 of the general synthetic route 1 are the same as those for the synthesis of compound 1003.

[0561] Step 3: Synthesize compound 6 of general synthetic route 5

[0562]

[0563] Compound 4 (500 mg), intermediate M (compound 5, 298.68 mg), reactant STAB (328.68 mg), and DCE (10 mL) were added to a three-necked flask. The reaction solution was stirred in N2 at 25 °C for 16 h. Then, 100 mL of water was added to the reaction solution, and the mixture was extracted twice with EA (100 mL). The organic phases were combined, concentrated, and filtered through a 1:20 DCM:MeOH column after adding silica gel. The sample was collected and concentrated to obtain compound 6 (500 mg).

[0564] Step 4: Synthesize compound 1114

[0565]

[0566] Compound 6 (450 mg) and solvent THF (5 mL) were added to a three-necked flask, followed by 3.65 mL of 1 M dioxane hydrochloride solution. After stirring at 25 °C for 2 h, the mixture was diluted with 50 mL of EA, and the organic phase was washed with 50 mL of saturated NaHCO3 solution and water. The organic layer was dried over anhydrous Na2SO4, filtered, concentrated, and then subjected to silica gel column chromatography (MeOH:DCM 1:20) to give compound 1114 (107 mg, yield 24.84%). 1 H NMR (400MHz, CDCl3) δ4.16-4.07(m,6H),3.60(t,J=4.8Hz,2H),2.62(t,J=7.0Hz,2H),2.49(s,3H),2.31(tt, J=8.4,5.5Hz,6H),1.70(d,J=4.6Hz,3H),1.64-1.39(m,18H),1.27(d,J=4.0Hz,78H),0.90(t,J=7.1Hz,18H).

[0567] Compound Z.1120 was synthesized according to general synthetic route 5.

[0568] Structural formula:

[0569] Chemical formula: C 72 H 142 N2O7

[0570] Molecular weight: 1147.90

[0571] Synthesized via general synthetic route 5, similar to the synthesis of compound 1114, except that intermediate C and 6-bromohexanal were used for compounds 2 and 5 in general synthetic route 5, respectively. ¹H NMR (400 MHz, Chloroform-d) δ 4.11 (s, 6H), 3.63 (s, 2H), 2.57 (t, J = 6.8 Hz, 4H), 2.37 (ddd, J = 8.5, 7.1, 4.3 Hz, 5H), 1.67–1.41 (m, 24H), 1.27 (s, 79H), 0.90 (t, J = 6.7 Hz, 18H).

[0572] Example 2: Exemplary Compounds and Their Properties

[0573] Exemplary compounds of the present invention and their properties are listed in Table 1, which were synthesized according to the process route in Example 1.

[0574] The c-pKa (molnetwork) and LogP (cLogP driver) values ​​of the exemplary compounds of this invention are calculated by the ChemDraw module of Chemoffice.

[0575] Table 1 Properties of Exemplary Compounds

[0576]

[0577]

[0578]

[0579]

[0580]

[0581]

[0582]

[0583]

[0584]

[0585]

[0586]

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603] The above properties of the exemplary compounds of the present invention indicate that they can serve as excellent surfactants, and are particularly suitable for preparing lipid nanoparticles for use as drug delivery carriers.

[0604] Example 3: Preparation of Nanoparticles

[0605] Luciferase mRNA was diluted in 10-100 mM pH 4.0 sodium citrate buffer solution to a concentration of 135 μg / ml. A lipid-mixed ethanol solution was prepared according to the following molar percentage ratio: DSPC: cholesterol: 2-[(polyethylene glycol)-2000]-N,N-tetracosylacetamide (ALC-0159) 50:10:38.5:1.5. The mRNA solution and lipid-mixed solution were mixed in a 3:1 volume ratio in a nanomedicine preparation device and then filtered through ultrafiltration to obtain luciferase mRNA lipid nanoparticles.

[0606] Example 4: Apparent pKa determination

[0607] The apparent pKa of lipid nanoparticles is correlated with their effectiveness in delivering nucleic acids (see Jayaraman et al., Angewandte Chemie, International Edition (2012), 51(34), 8529-8533; Semil et al., Nature Biotechnology 28, 172-176 (2010)). The apparent pKa of nanoparticles prepared from different lipid compounds was determined using fluorescence based on 2-(p-toluidine)-6-naphthalenesulfonic acid (TNS), a fluorescent probe that was non-fluorescent in water but became slightly fluorescent upon binding to a lipid membrane. Blank lipid nanoparticles were prepared according to the following molar ratio: DSPC:cholesterol:2-[(polyethylene glycol)-2000]-N,N-tetracosylacetamide (ALC-0159) in a ratio of 50:10:38.5:1.5. The nanoparticles were diluted in different pH buffers ranging from pH 2 to 11, mixed with TNS aqueous solution, and then added to 96-well plates (black background). The excitation wavelength was 325 nm and the emission wavelength was 435 nm. The top detection was performed, and each well was measured 10 times. The fluorescence intensity was recorded, and a pH-fluorescence intensity curve was plotted. The apparent pKa was measured. The apparent pKa data of the lipid nanoparticles prepared by each exemplary compound are shown in Table 2.

[0608] Example 5: Determination of nanoparticle size, zeta potential, and encapsulation efficiency

[0609] 1. Particle size and polydispersity index (PDI) determination: The average particle size and PDI of the nanoparticle sample solution in the examples were determined by dynamic light scattering using a Malvern ZetaSizer Nano ZS90 microscope. The measurement angle was 90°, the refractive index of the dispersant was 1.330, and the test temperature was 25°C.

[0610] 2. Zeta potential: The zeta potential of the nanoparticle sample solution in the examples was determined using a Malvern ZetaSizer Nano ZS90 based on electrophoretic light scattering (ELS) technology. The dispersant refractive index was 1.330, and the test temperature was 25℃.

[0611] 3. Encapsulation efficiency: The encapsulation efficiency of luciferase mRNA in lipid nanoparticles was determined using the Quant-it Ribogreen RNA Quantification Kit (ThermoFisher Scientific, UK) according to the manufacturer's instructions.

[0612] The average particle size, PDI, Zeta potential, and encapsulation efficiency of the lipid nanoparticles prepared from each exemplary compound are shown in Table 2.

[0613] Table 2. Summary of apparent pKa, particle size, PDI, Zeta potential, and encapsulation efficiency of lipid nanoparticles prepared from the compounds.

[0614] Compound numbering Apparent pKa Particle size (nm) Dispersion Index (PDI) Zeta potential (mV) Encapsulation efficiency (%) 1002 / 76.23 0.111 -13.3 / 1003 / 62.59 0.127 -3.64 95.4％ 1005 6.10 55.44 0.076 -0.105 91.7％ 1007 5.13 85.78 0.068 -4.98 95.9％ 1008 6.14 73.49 0.019 -6.06 95.1％ 1009 5.23 67.63 0.126 2.60 92.7％ 1011 6.30 60.38 0.193 2.31 91.8％ 1012 6.05 80.07 0.051 0.811 87.4％ 1013 5.75 82.41 0.123 -2.23 90.7％ 1014 / 85.4 0.111 3.06 70.3％ 1015 / 102.3 0.136 4.27 80.3％ 1025 / 77.74 0.072 2.84 64.5％ 1029 5.41 77.94 0.057 -3.71 90.6％ 1059 5.18 72.71 0.067 -10.0 93.0％ 1111 / 110.3 0.103 -0.902 83.2％ 1113 5.91 58.00 0.082 4.57 100.3％ 1118 5.11 108.1 0.051 -3.96 95.9％ 1120 6.14 98.18 0.049 -6.05 97.2％

[0615] Example 6: In vitro cell activity evaluation of lipid nanoparticles

[0616] The in vitro transfection efficiency of lipid nanoparticle compositions prepared with various exemplary compounds was evaluated using HEK-293T cells. HEK293T cells were routinely cultured in DMEM + 10% FBS medium to ensure they were in the logarithmic growth phase. One day before transfection, cells were seeded at an appropriate density into 96-well culture plates and allowed to grow overnight. At transfection, cell confluence should reach 70-90%. The lipid nanoparticle compositions prepared with each exemplary compound were diluted to four different concentrations using DMEM and added to 96-well cell culture plates, achieving concentrations of 400 ng, 200 ng, 100 ng, and 50 ng per well, respectively. A positive control (PC) was obtained by transfecting the luciferase plasmid with Lipofectamine 2000. After incubation at 37℃ and 5% CO2 for 24 h, substrate was added to the wells, and luciferase activity was measured using a microplate reader. The results are shown in Table 3. It can be seen that the luciferase mRNA lipid nanoparticles prepared using the exemplary compound expressed luciferase in HEK 293T cells (dose: 100 ng, 96-well plate) with a fluorescence intensity >10. 3 RLU indicates that the luciferase mRNA lipid nanoparticles prepared using the ionizable cationic lipid compounds of the present invention exhibit biological activity in in vitro cells and have low cytotoxicity. The luciferase activity of the luciferase mRNA lipid nanoparticles prepared with compound 1005 in HEK293T cells is shown in the appendix. Figure 1 It can be seen that, at different doses (50-400 ng), the fluorescence intensity of the prepared luciferase mRNA lipid nanoparticles expressing luciferase in HEK 293T cells is >10. 7 RLU was observed, and the fluorescence intensity increased with increasing dose, indicating that the expression level of luciferase increased accordingly with increasing dose. The prepared lipid nanoparticles have good mRNA delivery capability and safety.

[0617] Example 7: In vivo activity evaluation of lipid nanoparticles

[0618] The in vivo transfection efficiency and safety of lipid nanoparticle compositions prepared from each exemplary compound were evaluated using 6-8 week old female Balb / c mice. The lipid nanoparticles prepared from each exemplary compound were administered via tail vein injection at a single dose of 0.3 mpk. In vivo imaging of the animals was performed using the PerkinElmer small animal imaging system at specific time points after administration (e.g., 3h, 6h) to measure bioluminescent signals. The results are shown in Table 3. Whole-body in vivo imaging of mice 3h / 6h after intravenous administration of the luciferase mRNA lipid nanoparticles prepared from compound 1008 is shown in Appendix 1008. Figure 2 As is evident from Table 3, after 6 hours of administration of the lipid nanoparticles prepared with each of the exemplary compounds, the mean whole-body fluorescence signal in mice was greater than 10. 4 p / s / cm 2 The / sr result indicates that the luciferase mRNA lipid nanoparticles containing the exemplary compounds induced effective in vivo expression of luciferase. Furthermore, no abnormal activity or other toxic reactions were observed in mice after administration of the lipid nanoparticles prepared with the respective exemplary compounds. This demonstrates that the luciferase mRNA lipid nanoparticles prepared using the ionizable cationic lipid compounds of the present invention exhibit biological activity in animals and possess low toxicity.

[0619] Table 3 Summary of in vivo / in vitro activity evaluation of lipid nanoparticles prepared from the compounds

[0620]

[0621] The present invention is not limited to the technical means disclosed above, but also includes technical solutions composed of any combination of the above technical features. The above descriptions are specific embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications are also considered within the scope of protection of the present invention.

Claims

1. An ionizable cationic lipid compound, characterized in that, The ionizable cationic lipid compound is a compound of formula (1F), or its salt, stereoisomer, or tautomer: Among them, R1, R2, and R3 are H and C independently of each other. 5-40 Straight-chain or branched alkyl, C 5-40 Straight-chain or branched alkenyl, C 5-40 The side chains are: straight-chain or branched alkynyl groups, 3-6 membered saturated or partially unsaturated cyclic hydrocarbon groups containing 1-3 side chains, or 6-10 membered aromatic groups containing 1-3 side chains; wherein the side chains are independently selected from C... 10-30 Straight-chain or branched alkyl groups, C 10-30 Straight-chain or branched alkenyl, C 10-30 Straight-chain or branched alkynyl group; condition that at most one of R1, R2, and R3 is H; M is selected from -NR4R5, saturated or partially unsaturated 3-6 membered heterocyclic groups containing at least one nitrogen atom, and 6-10 membered heteroaryl groups containing at least one nitrogen atom, wherein the heterocyclic group or heteroaryl group is not substituted or is substituted by one or more -OH, carboxyl, amino, oxo, or halogen groups. R4 and R5 are H and C, respectively, which are independent of each other. 1-6 Straight-chain or branched alkyl groups, C 2-6 Straight-chain or branched alkenyl or C 2-6 Straight-chain or branched alkynyl groups, wherein the C 1-6 Straight-chain or branched alkyl groups, C 2-6 Straight-chain or branched alkenyl or C 2-6 The straight-chain or branched alkynyl group is either unsubstituted or substituted by one or more -OH, carboxyl, amino, amide, amidine, guanidine, or halogen groups; G1, G2, and G3 are independently -O-, -S-, -NR6-, -SS-, -C(=O)-, -C(=S)-, -C(=O)O-, -CH(OH)-, -OC(=O)-, -C(=O)NR6-, -NR6C(=O)-, -OC(=O)O-, -NR6C(=O)O-, -OC(=O)NR6-, -NR6C(=O)NR 13 -, -C(=O)S-, -C(=S)S-, -SC(=S)-, -SC(=O)-, -OC(=O)S-, -SC(=O)O-, -SC(=O)S-, -OS(=O)2O-, -S(=O)2O-, -OS (=O)2-, -S(=O)2-, -S(=O)2-NR6-, -NR6-S(=O)2-, -P(=O)(OR6)O-, -OP(=O)(OR6)- or -OP(=O)(OR6)O-; where each R6, R 13 They are independently selected from H, hydroxyl, and C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-30 Alkenes, whether straight-chain or branched; k is an integer from 1 to 6; R9, R 10 H is independent of each other; X2 is selected from -O-, -S-, NR 16 -, -SS-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR 16 -、 -NR 16 C(=O)-, -OC(=O)O-, -NR 16 C(=O)O-, -OC(=O)NR 16 -, -NR 16 C(=O)NR 17 -, -P(=O)(OR 16 )O-, -OP(=O)(OR 16 )-, or -OP(=O)(OR 16 )O-; R 11 R 12 The elements that are independent of each other are H, hydroxyl, halogen, and C. 1-6 Straight-chain or branched alkyl or cycloalkyl, C 2-6 Straight-chain or branched alkenyl groups, each R 16 R 17 They are selected independently from H and C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-30 Alkenes, whether straight-chain or branched; Among them, R4, R5, R6, R 11 R 12 R 13 R 16 R 17 The alkyl, cycloalkyl, and alkenyl groups described herein are either unsubstituted or substituted by one or more groups selected from hydroxyl, mercapto, amino, substituted amino, and halogen groups; the salts do not include quaternary ammonium salts.

2. The ionizable cationic lipid compound according to claim 1, characterized in that, R1, R2, and R3 are independently the following groups: Among them, Y does not exist, or it is C. 1-30 Straight-chain or branched alkyl or cycloalkyl, C 2-20 Straight-chain or branched alkenyl, C 2-20 Straight-chain or branched alkynyl group; R1' and R2' are independently H and C. 1-30 Straight-chain or branched alkyl groups, C 2-30 Straight-chain or branched alkenyl, C 2-30 Straight-chain or branched alkynyl groups, with a total carbon chain length of 8-40 for Y, R1' and R2'.

3. The ionizable cationic lipid compound according to claim 1, characterized in that, R1, R2, and R3 are independently selected from the following groups: Among them, R1' and R2' are H and C, respectively, which are independent of each other. 1-30 Straight-chain or branched alkyl groups, C 2-30 Straight-chain or branched alkenyl, C 2-30 Straight-chain or branched alkynyl groups, with a total carbon chain length of 8-30 for R1' and R2'.

4. The ionizable cationic lipid compound according to claim 1, characterized in that, R1, R2, and R3 are independently selected from any one of the following groups:

5. The ionizable cationic lipid compound according to claim 1, characterized in that, G1, G2, and G3 are independently -O-, -S-, -NR6-, -SS-, -C(=O)-, -C(=O)O-, -CH(OH)-, -OC(=O)-, -C(=O)NR6-, -NR6C(=O)-, -OC(=O)O-, -NR6C(=O)O-, -OC(=O)NR6-, -NR6C(=O)NR 13 -, -P(=O)(OR6)O-, -OP(=O)(OR6)- or -OP(=O)(OR6)O-.

6. The ionizable cationic lipid compound according to claim 1, characterized in that, M is selected from the following structures: Where m' and n' are independent integers from 0 to 6, and R1” and R2” are independent integers H and C. 1-6 alkyl, C 2-6 The nitrogen-containing heterocycle is selected from pyrrole, imidazole, pyridine, pyrazole, triazole, oxazole, isoxazole, thiophene, isothiazole, pyridazine, pyrazine, piperazine, indole, benzimidazole, carbazole, quinoline, isoquinoline, purine, and pyrimidine, and their tautomeric forms, and is not substituted or optionally is substituted by one or more groups selected from hydroxyl, mercapto, amino, substituted amino, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 6-14 The aryl group is substituted with an organic group.

7. The ionizable cationic lipid compound according to claim 1, characterized in that, The compounds of formula (1E) are selected from the compounds shown in formula (1H):

8. The ionizable cationic lipid compound of claim 1, selected from:

9. The method for preparing the ionizable cationic lipid compound according to any one of claims 1-8, characterized in that, This includes the steps of reacting compound (II) with compound (III): Wherein, Xa and Xb are groups containing leaving groups or nucleophilic groups, and Xa and Xb form L1 through nucleophilic reactions or condensation reactions; Where L1 is NH and L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - Other substituents are defined as in claims 1-6.

10. The method for preparing the ionizable cationic lipid compound according to any one of claims 1-8, characterized in that, This includes the steps of reacting compound (IV) with compound (V): Where Xc and Xd are groups containing leaving groups or nucleophilic groups, and Xc and Xd form L2 through nucleophilic reactions or condensation reactions; Where L1 is NH and L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - Other substituents are defined as in claims 1-6.

11. The method for preparing the ionizable cationic lipid compound according to any one of claims 1-8, characterized in that, This includes the steps of reacting the compound of formula (VI): Where Xe is a group containing a leaving group or a nucleophilic group, Xf is a compound containing a leaving group or a nucleophilic group, and Xe and Xf form M through a nucleophilic reaction or a condensation reaction; Where L1 is NH and L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - Other substituents are defined as in claims 1-6.

12. The method for preparing the ionizable cationic lipid compound according to any one of claims 1-8, characterized in that, The steps include reacting compound (VII) sequentially with compound (VIII), compound (IX), and compound (X): Wherein Xg, Xh, Xi, Xj, Xk, and Xl are groups containing leaving groups or nucleophilic groups, and Xg and Xj form G1 through nucleophilic reaction or condensation reaction, Xh and Xk form G2 through nucleophilic reaction or condensation reaction, and Xi and Xl form G3 through nucleophilic reaction or condensation reaction; Where L1 is NH and L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - Other substituents are defined as in claims 1-6.

13. The method for preparing the ionizable cationic lipid compound according to any one of claims 1-8, characterized in that, This includes the steps of reacting compound (XI) with compound (XII): The steps for the reaction of compound (XIII) with compound (XII): Where Xm is a group containing a nucleophilic group, and Xm has the same C=CL as the compound of formula (XII). 2a X1-L2 is formed through an addition reaction; Where X1 is NH and L2 is -(CR9R) 10 ) n -X2-(CR 11 R 12 ) k - Other substituents are defined as in claims 1-6.

14. The preparation method according to any one of claims 9-13, characterized in that... R1, R2, and R3 are independently the following groups: The meanings of Y, R1', and R2' are the same as in claim 2; This also includes the steps of forming the tail chains R1, R2, and R3: Where X is a leaving group.

15. The preparation method according to any one of claims 9-13, characterized in that, The raw materials used in the reaction process also contain protecting groups, and the reaction steps include protection and / or deprotection steps.

16. The use of the ionizable cationic lipid compound as described in any one of claims 1-8 as a surfactant.

17. The application as described in claim 16, characterized in that, The ionizable cationic lipid compounds are used as emulsifiers, suspending agents, dispersants, solubilizers, lubricants, thickeners, antibacterial agents, or preservatives.

18. The use of the ionizable cationic lipid compound as described in any one of claims 1-8 in the preparation of cosmetic compositions.

19. The use of the ionizable cationic lipid compound according to any one of claims 1-8 in the preparation of lipid compositions, characterized in that, The ionizable cationic lipid compound is used as a drug delivery carrier.

20. A lipid composition, characterized in that, The lipid component of the lipid composition includes the ionizable cationic lipid compound as described in any one of claims 1-8.

21. The lipid composition according to claim 20, characterized in that, The lipid composition further comprises other lipids, including phospholipids, structural lipids, and PEG-conjugated lipids; the molar ratio of the ionizable cationic lipid compound to other lipid components is 1:0.2-5; the active ingredient of the lipid nanoparticle composition includes therapeutic and / or preventive agents; the therapeutic and / or preventive agents are vaccines or compounds capable of inducing an immune response; the mass ratio of the lipid component to the active ingredient is 10:1 to 60:

1.

22. The lipid composition according to claim 21, characterized in that, The therapeutic and / or preventive agent is a nucleic acid; the nucleic acid is DNA or RNA.

23. The use of the lipid composition according to any one of claims 20-22 in a medicament for treating diseases in mammals, characterized in that, The disease is characterized by dysfunction or abnormal protein or polypeptide activity; the disease is selected from infectious diseases, cancer, proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases.

24. A pharmaceutical composition, characterized in that, It comprises the lipid composition according to any one of claims 20-22 and a pharmaceutically acceptable carrier.

25. The pharmaceutical composition according to claim 24, characterized in that, The pharmaceutical composition further includes a pharmaceutically active compound selected from anti-inflammatory compounds, steroids, statins, estradiol, BTK inhibitors, S1P1 agonists, glucocorticoid receptor modulators, and antihistamines.

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