Ionizable cationic lipid compound and use thereof

Abstract

An ionizable cationic lipid compound having a structure as represented by formula (I), which can be used for preparing a lipid nanoparticle (LNP) for the delivery of a therapeutic agent and / or a prophylactic agent. The LNP prepared with the ionizable cationic lipid compound has a better stability and transfection efficiency, and can efficiently and stably deliver a bioactive substance (comprising a nucleic acid, e.g., mRNA) to a target cell or organ, thereby eliciting a highly specific antibody response in vivo.

Description

Ionizable cationic lipid compound and its application Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to a cationic lipid compound and its application in the delivery of bioactive substances. Background Art

[0002] Nucleic acid drugs mainly refer to compounds containing nucleotide or deoxynucleotide structures with genetic characteristics and pharmacological activity. They can be used to treat tumors, tissue regeneration, wound healing, pulmonary fibrosis, inflammatory diseases, microbial infections, etc. After nucleic acid drugs are injected into the human body, an efficient and safe drug delivery system is required to deliver them to the lesion site. The drug delivery system needs to stay for a sufficient time to accurately target the lesion site while avoiding damage to normal cells.

[0003] Current delivery systems can be divided into viral vectors and non-viral vectors. Viral vectors are less used in nucleic acid drugs due to their immunogenicity, tumorigenicity, and limited drug loading capacity; non-viral vectors, such as polymers and lipids (liposomes or LNPs), can bind nucleic acid drugs to specific ligands to enable them to target specific cells, and are widely used in current nucleic acid drugs. LNP is one of the most widely used delivery systems for nucleic acid drug research. The LNP delivery system can safely and effectively deliver nucleic acids. It has advantages such as high nucleic acid encapsulation rate, ability to effectively transfect cells, strong tissue penetration, low cytotoxicity and immunogenicity, which are conducive to drug delivery. Compared with other delivery systems, it has huge advantages. Therefore, the LNP delivery system has broad development and application prospects.

[0004] In the prior art, LNP delivery systems are often composed of ionizable lipids (cationic lipids), steroids, neutral lipids, PEG-lipids, nucleic acid drugs and other ingredients. For example: Patent document AU2020325221A1 discloses a composition for delivering LNPs to target cells, including (i) ionizable lipids; (ii) sterols or other structural lipids; (iii) non-cationic auxiliary lipids or phospholipids; (iv) PEG lipids and (v) agents encapsulated in and / or associated with LNPs (such as nucleic acid molecules). These four components enhance the delivery efficiency of target cells in a specific ratio. Patent document WO2021 / 250263A1 discloses a composition including ionizable lipids, phospholipids, sterols, PEG lipids and one or more nucleic acids, including less than about 1 mol% of C14-PEG2000 lipids, and specific percentages of other lipids. Patent CN102712935B discloses a lipid particle comprising: a cationic lipid; a neutral lipid, a zwitterionic lipid or an anionic lipid; a PEG-lipid; a sterol and a nucleic acid, and assembling the above components into a lipid particle with a solid core, wherein the solid core can achieve a higher coating efficiency. Patent document WO 2021 / 055849A1 discloses a lipid with the following structure: This structure can improve its safety, effectiveness and specificity. Patent document WO2021 / 026358Al discloses a target cell delivery lipid nanoparticle (LNP): including (i) ionizable lipids (ii) sterols or other structural lipids; (iii) non-cationic auxiliary lipids or phospholipids; (iv) payload; (v) polyethylene glycol lipids, as a drug delivery system, taking into account both safety and effectiveness. In recent years, it has been discovered that introducing cholesterol into ionizable lipid compounds can also be used to deliver nucleic acid drugs. Patent document US7514099B2 discloses a cholesterol amino lipid compound CLinDMA The compound can be combined with phospholipids, cholesterol, and PEG lipids to form a four-component LNP or a five-component LNP with phospholipids, DMOBA lipids, cholesterol, and PEG lipids to deliver siRNA. Patent document CN112424214A discloses an ionizable cationic lipid compound formed by cholesterol and a linear olefin (3) This compound is combined with cholesterol, DPPC, DOPE, and DMG-PEG200 to construct LNPs for nucleic acid delivery. Each of the above compounds needs to be combined with three or even four different lipid excipients to form a nucleic acid drug delivery vector formulation, making the construction process relatively complex. The existing technology has an urgent need to optimize the structure of each component in LNP, especially the ionizable cationic lipids, to further obtain a safe, effective, stable, simple to construct, and applicable LNP delivery system for different routes of administration. Summary of the Invention

[0005] In one aspect, the present invention provides a lipid compound represented by formula (I),

[0006] or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0007] L1, L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0008] G1, G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0009] One of R1, R2 and R3 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by -O-, -S-, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl replacement;

[0010] At the same time, another one of R1, R2 and R3 is selected from a steroid group;

[0011] At the same time, the third one of R1, R2 and R3 is selected from -(R4) q -NR a R b、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy;

[0012] R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0013] R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl;

[0014] m, n and p are each independently selected from 1, 2 or 3;

[0015] q is selected from 0 or 1.

[0016] On the other hand, the present invention also provides a lipid nanoparticle comprising a lipid compound represented by formula (I) or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

[0017] In another aspect, the present invention also provides a pharmaceutical composition comprising the lipid nanoparticles described herein and a pharmaceutically acceptable carrier.

[0018] In another aspect, the present invention also provides a method of delivering a therapeutic and / or prophylactic agent, comprising administering the pharmaceutical composition described herein to a subject in need thereof.

[0019] On the other hand, the present invention also provides the use of the lipid compound represented by formula (I) described herein or its stereoisomers, tautomers, and pharmaceutically acceptable salts in the preparation of a therapeutic and / or preventive agent delivery system. Beneficial effects:

[0020] The three-component lipid nanoparticles prepared using the lipid compounds of the present invention, or their stereoisomers, tautomers, or pharmaceutically acceptable salts, have a simple process and exhibit excellent stability and transfection efficiency. The lipid nanoparticles can be used to deliver nucleic acids (e.g., mRNA) efficiently and stably to target cells or organs, eliciting high specific antibody and cellular immune responses in experimental animals with good safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] FIG1 shows the transfection efficiency of GFP-mRNA-LNP in HEK293T cells detected by fluorescence microscopy.

[0022] Figure 2 shows the in vivo delivery efficiency of SARS-CoV-2 S protein mRNA-LNP in mice.

[0023] Figure 3 shows the level of antigen-specific IFN-γ cell immune activation in PBMCs detected by ELISPOT after mice were immunized with SARS-CoV-2 S protein mRNA-LNP.

[0024] FIG4 shows the effect of mRNA-LNP on HEK293T cell proliferation. DETAILED DESCRIPTION

[0025] definition

[0026] As used in this specification, the following words and phrases are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0027] As used herein, the term "lipid nanoparticle," or "LNP," refers to a particle having a nanometer size, eg, 1 nm to 1,000 nm, which comprises one or more types of lipid molecules.

[0028] As used herein, the term "gene medicine" generally consists of a vector or delivery system containing an engineered gene construct, the active ingredient of which may be DNA, RNA, genetically modified viruses, bacteria or cells. By introducing exogenous genes into target cells or tissues, it replaces, compensates, blocks or corrects specific genes to achieve the purpose of treating and preventing diseases.

[0029] As used herein, the term "nucleic acid" refers to a polymer containing at least two deoxyribonucleotides or ribonucleotides in single- or double-stranded form, and includes DNA, RNA, and hybrids thereof.

[0030] As used herein, the term "lipid compound" refers to a group of organic compounds, which include but are not limited to esters of fatty acids and are generally characterized by being poorly soluble in water but soluble in many organic solvents. The organic solvents of the present invention include but are not limited to: benzene, toluene, pentane, hexane, methanol, ethanol, isopropanol, ether, ethyl acetate, acetone, carbon tetrachloride.

[0031] As used herein, the term "alkyl" refers to a monovalent group having a straight or branched saturated hydrocarbon chain of 1 to 20 carbon atoms, more typically 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 6 carbon atoms. This term is exemplified by groups such as methyl, ethyl, 1-propyl (n-propyl), 2-propyl (isopropyl), 1-butyl (n-butyl), 2-methyl-1-propyl (isobutyl), 2-butyl (sec-butyl), 2-methyl-2-propyl (tert-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, 1-heptyl, 1-octyl, 1-nonyl, 1-decyl, and the like.

[0032] As used herein, the term "alkylene" refers to a divalent group having a straight or branched saturated hydrocarbon chain of 1 to 20 carbon atoms, more typically 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 6 carbon atoms. The term is exemplified by groups such as methylene, ethylene, propylene, butylene, pentylene, hexylene, and the like.

[0033] As used herein, the term "alkenyl" refers to a linear or branched unsaturated hydrocarbon chain monovalent group having 2 to 20 carbon atoms (more typically 2 to 10 carbon atoms, 2 to 8 carbon atoms, or 2 to 6 carbon atoms) and having carbon-carbon double bonds (e.g., 1, 2, or 3 carbon-carbon double bonds). The unsaturated carbon-carbon double bond can be present at any stable point along the chain. The term is exemplified by groups such as vinyl (i.e., -CH=CH2), propen-1-yl (i.e., -CH=CHCH3), propen-3-yl (or allyl, i.e., -CH2CH=CH2), propen-2-yl (i.e., -C(CH3)=CH2), butadienyl (including 1,2-butadienyl and 1,3-butadienyl), and the like.

[0034] As used herein, the term "alkenylene" refers to a divalent group of a straight or branched unsaturated hydrocarbon chain having 2 to 20 carbon atoms (more typically 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 6 carbon atoms) and having carbon-carbon double bonds (e.g., 1, 2, or 3 carbon-carbon double bonds). The unsaturated carbon-carbon double bond can be present at any stable point along the chain. The term is exemplified by groups such as ethenylene, propenylene, butenylene, pentenylene, hexenylene, and the like.

[0035] As used herein, the term "alkynyl" refers to a linear or branched unsaturated hydrocarbon chain monovalent group having 2 to 20 carbon atoms (more typically 2 to 10 carbon atoms, 2 to 8 carbon atoms, or 2 to 6 carbon atoms) and having carbon-carbon triple bonds (e.g., 1, 2, or 3 carbon-carbon triple bonds). The term is exemplified by groups such as ethynyl (i.e., -C≡CH), propargyl (i.e., -CH2C≡CH), propynyl (i.e., -C≡CCH3), and the like.

[0036] As used herein, the term "alkynylene" refers to a divalent group of a straight or branched unsaturated hydrocarbon chain having 2 to 20 carbon atoms (more typically 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 6 carbon atoms) and having carbon-carbon triple bonds (e.g., 1, 2, or 3 carbon-carbon triple bonds). The unsaturated carbon-carbon triple bond can exist at any stable point along the chain. The term is exemplified by groups such as ethynylene, propynylene, butynylene, pentynylene, hexynylene, and the like.

[0037] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0038] As used herein, the term "alkoxy" refers to an "alkyl-O-" group, wherein alkyl is as defined herein. This term is exemplified by groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

[0039] As used herein, the term "acyl" refers to "alkyl-C(=O)-", "alkenyl-C(=O)-", "alkynyl-C(=O)-", "aryl-C(=O)-", "heteroaryl-C(=O)-", "carbocyclyl-C(=O)-", "heterocyclyl-C(=O)-" groups, wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocyclyl, and heterocyclyl are as defined herein. This term is exemplified by groups such as formyl, acetyl, propionyl, n-butyryl, isobutyryl, n-valeryl, n-hexanoyl, acryloyloxy, benzoyl, cyclopropylacyl, and the like.

[0040] As used herein, the term "acyloxy" refers to "alkyl-C(=O)O-", "alkenyl-C(=O)O-", "alkynyl-C(=O)O-", "aryl-C(=O)O-", "heteroaryl-C(=O)O-", "carbocyclyl-C(=O)O-", "heterocyclyl-C(=O)-" groups, wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocyclyl, and heterocyclyl are as defined herein. This term is exemplified by groups such as formyloxy, acetyloxy, propionyloxy, n-butyryloxy, isobutyryloxy, n-pentanoyloxy, n-hexanoyloxy, and the like.

[0041] As used herein, the term "aryl" refers to an aromatic carbocyclic group of 6 to 14 carbon atoms (more typically 6 to 10 carbon atoms, or 6 carbon atoms) having a single ring (e.g., phenyl) or multiple rings (e.g., biphenyl) or multiple condensed (fused) rings (e.g., naphthyl, fluorenyl, and anthracenyl). The term is exemplified by groups such as phenyl, fluorenyl, naphthyl, anthracenyl, 1,2,3,4-tetrahydronaphthalene (if the point of attachment is through the aryl group), and the like.

[0042] As used herein, term " carbocyclic radical " refers to have 3 to 14 carbon atoms (more typically have 3 to 8 carbon atoms, or 3 to 6 carbon atoms) as the monocycle of annular atoms or the monoradical saturation or part unsaturated group of a plurality of thick (condensed) rings or bridged rings or spirocycles.Carbocyclic ring or carbocyclic radical can be saturated or partly unsaturated, and can be condensed with another saturated, partly unsaturated or aromatic ring, and condition is that the annular atoms being connected with target molecule is not aromatic carbon.The example of carbocyclic ring or carbocyclic radical includes, but is not limited to cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclopentadiene etc.

[0043] As used herein, the term "heteroaryl" refers to an aromatic ring group comprising a monocyclic or polycyclic fused ring (e.g., comprising 2 or 3 rings) having 5 to 14 ring atoms (more typically 5 to 10 ring atoms, or 5 to 6 ring atoms) in the ring, wherein in addition to carbon atoms, the ring atoms further comprise at least one heteroatom selected from oxygen, nitrogen and / or sulfur. If the ring is aromatic, the sulfur and nitrogen atoms may also exist in oxidized form. The polycyclic fused heteroaryl is a monocyclic heteroaryl as defined above fused with one or more rings selected from the following to form a polycyclic fused ring system: heteroaryl (to form, for example, naphthyridinyl, such as 1,8-naphthyridinyl), heterocycle (to form, for example, 1,2,3,4-tetrahydronaphthyridinyl, such as 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycle (to form, for example, 5,6,7,8-tetrahydroquinolinyl) and aryl (to form, for example, indazolyl). It will be understood that the point of attachment of the heteroaryl group can be at any suitable atom of the heteroaryl group, including carbon atoms and heteroatoms (eg, nitrogen). Exemplary heteroaryl groups include, but are not limited to, pyridinyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furanyl, oxadiazolyl, thiadiazolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl, quinazolinyl, 5,6,7,8-tetrahydroisoquinolinyl, benzofuranyl, benzimidazolyl, thiaindenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazolyl, and 3b,4,4a,5-tetrahydro-1H-cyclopropane[3,4]cyclopenta[1,2-c]pyrazolyl.

[0044] As used herein, the term "heterocyclyl" refers to a monocyclic or multiple condensed (fused) ring or bridged ring or spirocyclic ring having 3 to 14 ring atoms (more typically 3 to 10 ring atoms, or 3 to 6 ring atoms) in the ring, wherein the ring atoms also include at least one or more nitrogen atoms in addition to carbon atoms. The example of a heterocyclyl group includes, but is not limited to, an aziridine ring, an azetidine ring, a tetrahydropyrrole ring, a piperidine ring, an azepane ring, an azooctane ring, a tetrahydroimidazole ring, a tetrahydropyrazole ring, a tetrahydrooxazole ring, a tetrahydroisoxazole ring, a tetrahydrothiazole ring, a tetrahydroisothiazole ring, a piperazine ring, a morpholine ring, a dihydropyridyl, 4,5,6,7-tetrahydro-1H-benzo [d] imidazole, 4,5,6,7-tetrahydro-1H-imidazo [4,5-c] pyridine, etc. The nitrogen heterocyclic group in the present invention is a heterocyclic group containing a nitrogen atom in its structure, including but not limited to substituted or unsubstituted: aziridinyl, azetidinyl, β-propiolactam, pyrrolyl, piperidinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, caprolactam, pyranyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, piperazinyl, indolyl, benzimidazolyl, carbazolyl, quinolyl, isoquinolyl, pteridinyl, acridinyl, 7H-purinyl, phenazinyl, phenothiazinyl or 1H-azepinyl.

[0045] As used herein, the term "optionally substituted" means unsubstituted or substituted with one or more groups selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, halogen, hydroxy, cyano, nitro, amino, C3-C6 cycloalkyl, and oxo.

[0046] As used herein, the term "steroid" is an organic compound having a four-ring carbon skeleton structure as shown below.

[0047] Steroids include naturally occurring or synthetic steroids and their analogs. Steroids or their analogs include sterols or their analogs derived from plants and / or animals. Examples of steroids described herein include, but are not limited to, avenasterol, β-sitosterol, brassicasterol, ergocalciferol, campesterol, cholestanol, coprostanol, dehydrocholesterol, streptosterol, dihydroergocalciferol, cholesterol, dihydrocholesterol, dihydroergosterol, black sea sterol, epicholesterol, ergosterol, fucoxosterol, hexahydroluminosterol, hydroxycholesterol, luminosterol, algaesterol, sitostanol, stigmasterol, stigmasterol, cholic acid, glycocholic acid, taurocholic acid, deoxycholic acid, lithocholic acid, ent-cholesterol, epicholesterol, desmosterol, cholestanol, cholestanone, choletenone, 3p-[N-(N'N'-dimethylaminoethyl)carbamoylcholesterol (DC-Ch ol), 24(S)-hydroxycholesterol, 25-hydroxycholesterol, 25(R)-27-hydroxycholesterol, 22-oxa-cholesterol, 23-oxa-cholesterol, 24-oxa-cholesterol, cyclohexyl alcohol, 22-ketosterol, 20-hydroxysterol, 7-hydroxycholesterol, 19-hydroxycholesterol, 22-hydroxycholesterol, 25-hydroxycholesterol, 7-dehydrocholesterol, dehydroergosterol, dehydroepiandrosterone, lanosterol, dihydrolanosterol, lumiesterol, cetocalciferol, calcipotriol, coprostol, cholecalciferol, lupeol, ergocalciferol, 22-dihydroautocalciferol, tomatine, ursolic acid, chenodeoxycholic acid, zymosterol, diosgenin, etc.

[0048] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to effect treatment, as defined below, when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending on the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the mode of administration, etc., and can be readily determined by one of ordinary skill in the art.

[0049] As used herein, the term "stereoisomer" refers to a compound that has the same chemical composition and connectivity, but whose atoms have different orientations in space that cannot be interchanged by rotation about a single bond. "Stereoisomer" includes "diastereomers" and "enantiomers." "Diastereomers" refers to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral characteristics, and reactivity. Diastereomeric mixtures can be separated under high-resolution analytical procedures such as crystallization, electrophoresis, and chromatography. "Enantiomers" refers to two stereoisomers of a compound that are non-overlapping mirror images of each other.

[0050] As used herein, the term "tautomer" refers to the coexistence of two (or more) compounds that differ only in the position and electron distribution of one (or more) mobile atoms, such as keto-enol tautomers.

[0051] As used herein, the term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of a given compound and are not biologically or otherwise undesirable. Pharmaceutically acceptable salts can be acid addition salts and / or base addition salts. Acid addition salts can be prepared from inorganic acids and organic acids. Salts derived from inorganic acids include hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, bisulfate, hydrogenphosphate, dihydrogenphosphate, bicarbonate, etc.; salts derived from organic acids include formate, acetate, propionate, glycolate, pyruvate, oxalate, malate, malonate, succinate, maleate, fumarate, tartrate, citrate, benzoate, cinnamate, mandelate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate, lactate, nicotinate, lauryl sulfate, naphthalenesulfonate, camphorsulfonate, gluconate, glucuronate, oleate, palmitate, stearate, pamoate, trifluoroacetate, etc. Base addition salts can be formed with inorganic or organic bases. Salts derived from inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, lithium, barium, aluminum salts and the like; salts derived from organic bases include salts formed with various primary, secondary and tertiary amines, for example, ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, lysine, piperazine, piperidine, morpholine, tromethamine, choline and the like.

[0052] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0053] As used herein, the term "delivery system" refers to a formulation or composition that regulates the spatial, temporal, and dosage distribution of a biologically active ingredient in an organism.

[0054] Compound

[0055] In some embodiments, the present invention provides a lipid compound represented by formula (I)

[0056] or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L1, L2, L3, G1, G2, G3, R1, R2, R3, m, n, and p are as defined above.

[0057] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0058] L1 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0059] Each occurrence of G1 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0060] R1 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl replacement;

[0061] L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0062] G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NRa -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0063] One of R2 and R3 is selected from a steroid group, while the other is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy;

[0064] R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0065] R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl;

[0066] m, n and p are each independently selected from 1, 2 or 3;

[0067] q is selected from 0 or 1.

[0068] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0069] L1 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0070] Each occurrence of G1 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0071] R1 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl replacement;

[0072] L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0073] G2 is independently selected at each occurrence from -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0074] R2 is selected from a steroid group;

[0075] L3 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0076] G3 is independently selected at each occurrence from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0077] R3 is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C20 Alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy;

[0078] R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0079] R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl;

[0080] m, n and p are each independently selected from 1, 2 or 3;

[0081] q is selected from 0 or 1.

[0082] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0083] L1 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0084] Each occurrence of G1 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0085] R1 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl replacement;

[0086] L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0087] G2 is independently selected at each occurrence from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0088] R2 is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C20 acyloxy;

[0089] L3 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0090] G3 is independently selected from -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0091] R3 is selected from a steroid group;

[0092] R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0093] R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl;

[0094] m, n and p are each independently selected from 1, 2 or 3;

[0095] q is selected from 0 or 1.

[0096] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0097] L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0098] Each occurrence of G2 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0099] R2 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, carbocyclyl, aryl, heteroaryl, or heterocyclyl;

[0100] L1 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkynylidene, optionally substituted C1-C 20 acyl group;

[0101] G1 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(ORa )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0102] One of R1 and R3 is selected from a steroid group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy;

[0103] R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0104] R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl, optionally substituted C3-C 14 Carbocyclyl, optionally substituted C6-C 14 aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl;

[0105] m, n and p are each independently selected from 1, 2 or 3;

[0106] q is selected from 0 or 1.

[0107] In some embodiments, the present invention provides a lipid compound represented by formula (II)

[0108] or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L1, L2, L3, G1, G2, G3, R1, R2, R3, m, n, and p are as defined above.

[0109] In some embodiments, the steroid compound in the steroid compound group of the present invention is selected from naturally occurring steroid compounds or their analogs; preferably, it includes plant sterols and animal sterols, or their analogs; more preferably, it is selected from avenasterol, β-sitosterol, brassicasterol, ergocalciferol, campesterol, cholestanol, coprostanol, dehydrocholesterol, streptosterol, dihydroergocalciferol, cholesterol, dihydrocholesterol, dihydroergosterol, melanosterol, epicholesterol, ergosterol, fucoxosterol, hexahydroluminosterol, hydroxycholesterol, luminosterol, alginosterol, sitostanol, stigmasterol, stigmasterol, cholic acid, glycocholic acid, taurocholic acid, deoxycholic acid, lithocholic acid, ent-cholesterol, epicholesterol, demosterol, cholestanol, cholestanone, choletenone, 3p-[ N-(N'N'-dimethylaminoethyl)carbamoylcholesterol (DC-Chol), 24(S)-hydroxycholesterol, 25-hydroxycholesterol, 25(R)-27-hydroxycholesterol, 22-oxa-cholesterol, 23-oxa-cholesterol, 24-oxa-cholesterol, cyclohexyl alcohol, 22-ketosterol, 20-hydroxysterol, 7-hydroxycholesterol, 19-hydroxycholesterol, 22-hydroxycholesterol, 25-hydroxycholesterol, 7-dehydrocholesterol, dehydroergosterol, dehydroepiandrosterone, lanosterol, dihydrolanosterol, lumilastol, cetocalciferol, calcipotriol, coprostol, cholecalciferol, lupeol, ergocalciferol, 22-dihydroautocalciferol, tomatine, ursolic acid, chenodeoxycholic acid, zymosterol, diosgenin, etc.

[0110] In some embodiments, in the lipid compound provided by the present invention or its stereoisomers, tautomers, and pharmaceutically acceptable salts, the steroid compound in the steroid compound group is selected from cholesterol and cholesterol derivatives.

[0111] In some embodiments, in the lipid compounds provided herein or their stereoisomers, tautomers, and pharmaceutically acceptable salts, the steroidal compound group has the following structure:

[0112] R5 is selected from hydrogen, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxycarbonyl C1-C 20 alkyl-;

[0113] R6 is selected from hydrogen, halogen, cyano, hydroxy, amino, oxo, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl;

[0114] m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0115] In some embodiments, the steroid group is selected from:

[0116] Where R' is C 1- C 20 alkyl.

[0117] In some embodiments, the present invention provides a lipid compound represented by formula (IV-a) or formula (IV-b):

[0118] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, wherein L1, L2, L3, G1, G2, G3, R1, R2, R3, m, n, and p are as defined herein.

[0119] In some embodiments, the present invention provides a lipid compound represented by Formula (Va) or Formula (Vb):

[0120] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, wherein L1, L2, L3, G1, G2, G3, R1, R2, and R3 are as defined herein.

[0121] In some embodiments, the present invention provides a lipid compound represented by formula (VI-a) or formula (VI-b):

[0122] or its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein

[0123] L1, L2, L3, G1, G2, G3, R1, R2, and R3 are as defined herein.

[0124] In some specific embodiments, each occurrence of L1 is independently selected from optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Acyl; preferably, L1 is selected from optionally substituted C1-C6 alkylene, optionally substituted C2-C6 acyl;

[0125] Each occurrence of G1 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(Ra )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; preferably, G1 is selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-、-N(R a )C(=O)O-、-OC(=O)N(R a )-; More preferably, G1 is selected from -C(=O)O-, -OC(=O)-;

[0126] R a and R b Each independently selected from H, C1-C 20 Alkyl, C1-C 20 Alkenyl, C1-C 20 Alkynyl, carbocyclyl, aryl, heteroaryl, heterocyclyl; preferably, R a and R b Each independently selected from H, C1-C 20 Alkyl, C1-C 20 Alkenyl, C1-C 20 Alkynyl; more preferably, R a and R b Each independently selected from H, C1-C 20 alkyl;

[0127] R1 is selected from optionally substituted C1-C 20 Alkyl; wherein the C1-C 20 One or more -CH2- groups in the alkyl group may be optionally replaced by O, S, or a C3-C6 carbocyclic group.

[0128] In some specific embodiments, each occurrence of L1 is independently selected from optionally substituted methylene, ethylene, propylene, butylene, pentylene, hexylene, acetyl, propionyl, butyryl, pentanoyl, hexanoyl;

[0129] Each occurrence of G1 is independently selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(Ra )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-;

[0130] R a and R b Each independently selected from H, C1-C6 alkyl;

[0131] R1 is selected from

[0132] In some specific embodiments, L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 acyl group;

[0133] G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0134] One of R2 and R3 is selected from a cholesterol group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -5 or 6-membered nitrogen-containing heteroaryl, -(R4) q-5 or 6-membered nitrogen-containing heterocyclic group; wherein the 5 or 6-membered nitrogen-containing heteroaryl group and the 5 or 6-membered nitrogen-containing heterocyclic group are optionally substituted by one or more groups selected from the following: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogen, hydroxyl, thiol, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy;

[0135] R4 is selected from C1-C6 alkylene;

[0136] R a and R b Each independently selected from H, C1-C6 alkyl;

[0137] q is selected from 0 or 1.

[0138] In some specific embodiments, L2 and L3 are each independently selected from a bond, an optionally substituted C1-C6 alkylene, an optionally substituted C2-C6 acyl group;

[0139] G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -;

[0140] One of R2 and R3 is selected from a cholesterol group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -5 or 6-membered nitrogen-containing heteroaryl, -(R4) q -5 or 6-membered nitrogen-containing heterocyclic group; wherein the 5 or 6-membered nitrogen-containing heteroaryl group and the 5 or 6-membered nitrogen-containing heterocyclic group are optionally substituted by one or more groups selected from the following: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogen, hydroxyl, thiol, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy;

[0141] R4 is selected from C1-C6 alkylene;

[0142] R a and R b Each independently selected from H, C1-C6 alkyl;

[0143] n is selected from 0 or 1.

[0144] In some embodiments, the present invention provides a compound selected from the group consisting of compounds shown in Table 1 or a pharmaceutically acceptable salt thereof.

[0145] Lipid nanoparticles (LNPs)

[0146] In some embodiments, the present invention provides a lipid nanoparticle as a delivery vehicle for therapeutic and / or prophylactic agents (e.g., nucleic acids including DNA, RNA, etc.), comprising a lipid compound as described herein or its stereoisomers, tautomers, and pharmaceutically acceptable salts thereof. In some embodiments, the lipid nanoparticles described herein further comprise one or more phospholipids. In some embodiments, the lipid nanoparticles described herein further comprise one or more PEG lipids. In some embodiments, the lipid nanoparticles described herein further comprise a combination of phospholipids and PEG lipids. The lipid nanoparticles described herein can deliver therapeutic and / or prophylactic agents to target sites of interest (e.g., cells, tissues, organs, etc.). Therefore, the lipid nanoparticles described herein further comprise one or more therapeutic or prophylactic agents (e.g., nucleic acids, particularly therapeutic nucleic acids (TNA)).

[0147] In some embodiments, the lipid nanoparticles have a molar ratio of lipid compound: phospholipid: PEG lipid of 30-90:10-60:0.5-20; preferably, the molar ratio of lipid compound: phospholipid: PEG lipid is 30-80:30-80:0.5-20; more preferably, the molar ratio of lipid compound: phospholipid: PEG-lipid is 40-60:40-60:0.5-5; most preferably, the molar ratio of lipid compound: phospholipid: PEG-lipid is 49.25:49.25:1.5.

[0148] phospholipids

[0149] In some embodiments, the lipid nanoparticles described herein further comprise a phospholipid. Examples of phospholipids include, but are not limited to, distearoyl-sn-glycero-phosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoylphosphatidylcholine (DOPG), dioleoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylcholine (DP ... Phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine (e.g., 16-O-monomethyl PE), dimethylphosphatidylethanolamine (e.g., 16-O-dimethyl PE), 18-1-trans PE, 1-stearoyl-2-oleoylphosphatidylethanolamine Ethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC), dioleoylphosphatidylserine (DOPS), sphingomyelin (SM), dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DSPG), rutoylphosphatidylcholine (DEPC), palmitoyloleoylphosphatidylglycerol (POPG), dioleoyl-phosphatidylethanolamine (DEPE) The phospholipids of claim 1 are phospholipids, phosphatidylcholine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebroside, hexadecyl phosphate, lysophosphatidylcholine, dilinoleoylphosphatidylcholine or its mixture. It should be understood that other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl group in these lipids is preferably an acyl group derived from a fatty acid with a C10-C24 carbon chain, such as lauroyl, myristoyl, palmitoyl, stearyl or oleoyl.

[0150] In some embodiments, the molar percentage of phospholipids in the total lipid of the lipid nanoparticles is about 15% to about 65%, for example, about 20% to about 65%, about 25% to about 65%, about 30% to about 65%, about 35% to about 65%, about 40% to about 65%, about 45% to about 65%, about 50% to about 65%, about 55% to about 65%, about 60% to about 65%, about 15% to about 20%, about 20% to about 25%, about 25% to about 30%, about 30% to about 35%, about 35% to about 40%, about 40% to about 45%, about 45% to about 50%, about 50% to about 55%, about 55% to about 60%, about 60% to about 65%.

[0151] PEG lipids

[0152] In some embodiments, PEG lipids are incorporated into lipid nanoparticles as described herein to inhibit aggregation of particles, thereby improving the stability of lipid nanoparticles. In some embodiments, PEG lipids described herein are lipids that are covalently or non-covalently linked to one or more polyethylene glycol (PEG) chains. In some embodiments, PEG lipids described herein are lipids that are covalently linked to one or more polyethylene glycol (PEG) chains.

[0153] In some embodiments, the molecular weight of PEG molecules suitable for use in the PEG lipids described herein is from about 500 to about 10,000, from about 1,000 to about 10,000, from about 1,000 to about 5,000, from about 1,000 to about 4,000, from about 1,000 to about 3,000, from about 1,000 to about 2,000, e.g., PEG 2000, PEG 2500, PEG 3000, etc.

[0154] Examples of PEG lipids include, but are not limited to, PEG-diacylglycerols (DAG) (e.g., 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG)), PEG-dialkoxypropyl (DAA), PEG-phospholipids, PEG-ceramide (Cer), PEGylated phosphatidylethanolamine (PEG-PE), PEG succinate diacylglycerols (PEGS-DAG) (e.g., 4-O-(2',3'-di-tetradecanoyloxy) )propyl-1-O-(w-methoxy(polyethoxy)ethyl)succinate (PEG-S-DMG)), PEG dialkoxypropylaminoformamide, sodium N-(carbonyl-methoxypolyethylene glycol 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, PEG-dilauroyloxypropyl, PEG-dimyristoyloxypropyl, PEG-dipalmitoyloxypropyl, PEG-distearoyloxypropyl, 1-(monomethoxy-polyethylene glycol)-2,3 -Dimyristoylglycerol-PEG (DMG-PEG), distearoyl-rac-glycerol-PEG (DSG-PEG), PEG-dilauroylglycerol, PEG-dipalmitoylglycerol, PEG-distearoylglycerol, PEG-dilauroylglyceramide, PEG-dimyristoylglyceramide, PEG-dipalmitoylglyceramide, PEG-distearoylglyceramide, (1-[8'-(cholest-5-ene-3β-oxy)carboxamido-3', 6′-dioxaoctyl]carbamoyl-ω-methyl-poly(ethylene glycol) (PEG-cholesterol), 3,4-ditetradecyloxybenzyl-ω-methyl-poly(ethylene glycol) ether (PEG-DMB), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)] (DSPE-PEG), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-poly(ethylene glycol)-hydroxy (DSPE-PEG-OH).

[0155] In some embodiments, the molar percentage of PEG lipid in the total lipid of the lipid nanoparticle is about 0.1% to about 10%, for example, about 1.0% to about 10%, about 1.5% to about 10%, about 2.0% to about 10%, about 2.5% to about 10%, about 3.0% to about 10%, about 3.5% to about 10%, about 4.0% to about 10%, about 4.5% to about 10%, about 5.0% to about 10%, about 5.5% to about 10%, about 6.0% to about 10%, about 6.5% to about 10%, about 7.0% to about 10%, about 7.5% to about 10%, about 8.0% to about 10%, about 8.5% to about 10%, about 9. 9.0% to about 10%, about 9.5% to about 10%, about 1.0% to about 1.5%, about 1.5% to about 2.0%, about 2.0% to about 2.5%, about 2.5% to about 3.0%, about 3.0% to about 3.5%, about 3.5% to about 4.0%, about 4.0% to about 4.5%, about 4.5% to about 5.0%, about 5.0% to about 5.5%, about 5.5% to about 6.0%, about 6.0% to about 6.5%, about 6.5% to about 7.0%, about 7.0% to about 7.5%, about 7.5% to about 8.0%, about 8.0% to about 8.5%, about 8.5% to about 9.0%, about 9.0% to about 10%.

[0156] Particle size of lipid nanoparticles

[0157] In some embodiments, the lipid nanoparticles described herein have a particle size range of about 40 nm to about 150 nm, such as about 45 nm to about 150 nm, about 50 nm to about 150 nm, about 55 nm to about 150 nm, about 60 nm to about 150 nm, about 65 nm to about 150 nm, about 70 nm to about 150 nm, about 75 nm to about 150 nm, about 80 nm to about 150 nm, about 85 nm to about 150 nm, about 90 nm to about 150 nm, about 95 nm to about 150 nm, about 100 nm to about 150 nm, about 105 nm to about 150 nm, about 110 nm to about 150 nm, about 115 nm to about 150 nm, about 120 nm to about 150 nm, about 125 nm to about 150 nm, about 130 nm to about 150 nm, about 135 nm to about 150 nm, about 140 nm to about 150 nm, about 145 nm to about 150 nm. In some embodiments, the lipid nanoparticles described herein have a particle size range of about 40 nm to about 120 nm, e.g., about 45 nm to about 120 nm, about 50 nm to about 120 nm, about 55 nm to about 120 nm, about 60 nm to about 120 nm, about 65 nm to about 120 nm, about 70 nm to about 120 nm, about 75 nm to about 120 nm, about 80 nm to about 120 nm, about 85 nm to about 120 nm, about 90 nm to about 120 nm, about 95 nm to about 120 nm, about 100 nm to about 120 nm, about 105 nm to about 120 nm, 110 nm to about 120 nm, 115 nm to about 120 nm. In some embodiments, the lipid nanoparticles described herein have a particle size range of about 40 nm to about 110 nm, for example, about 45 nm to about 110 nm, about 50 nm to about 110 nm, about 55 nm to about 110 nm, about 60 nm to about 110 nm, about 65 nm to about 110 nm, about 70 nm to about 110 nm, about 75 nm to about 110 nm, about 80 nm to about 110 nm, about 85 nm to about 110 nm, about 90 nm to about 110 nm, about 95 nm to about 110 nm, about 100 nm to about 110 nm, about 105 nm to about 110 nm. In some embodiments, the lipid nanoparticles described herein have a particle size range of about 40 nm to about 100 nm, for example, about 45 nm to about 100 nm, about 50 nm to about 100 nm, about 55 nm to about 100 nm, about 60 nm to about 100 nm, about 65 nm to about 100 nm, about 70 nm to about 100 nm, about 75 nm to about 100 nm, about 80 nm to about 100 nm, about 85 nm to about 100 nm, about 90 nm to about 100 nm, about 95 nm to about 100 nm.In some embodiments, the particle size of the lipid nanoparticles described herein ranges from about 40 nm to about 90 nm, for example, about 45 nm to about 90 nm, about 50 nm to about 90 nm, about 55 nm to about 90 nm, about 60 nm to about 90 nm, about 65 nm to about 90 nm, about 70 nm to about 90 nm, about 75 nm to about 90 nm, about 80 nm to about 90 nm, about 85 nm to about 90 nm. In some embodiments, the particle size of the lipid nanoparticles described herein ranges from about 40 nm to about 85 nm, for example, about 45 nm to about 85 nm, about 50 nm to about 85 nm, about 55 nm to about 85 nm, about 60 nm to about 85 nm, about 65 nm to about 85 nm, about 70 nm to about 85 nm, about 75 nm to about 85 nm, about 80 nm to about 85 nm. In some embodiments, the particle size range of the lipid nanoparticles described herein is from about 40 nm to about 80 nm, for example, from about 45 nm to about 80 nm, from about 50 nm to about 80 nm, from about 55 nm to about 80 nm, from about 60 nm to about 80 nm, from about 65 nm to about 80 nm, from about 70 nm to about 80 nm, from about 75 nm to about 80 nm. In some embodiments, the particle size range of the lipid nanoparticles described herein is from about 40 nm to about 70 nm, for example, from about 45 nm to about 70 nm, from about 50 nm to about 70 nm, from about 55 nm to about 70 nm, from about 60 nm to about 70 nm, from about 65 nm to about 70 nm. In some embodiments, the particle size range of the lipid nanoparticles described herein is from about 40 nm to about 60 nm, for example, from about 45 nm to about 60 nm, from about 50 nm to about 60 nm, from about 55 nm to about 60 nm.

[0158] Lipid / nucleic acid ratio

[0159] In some embodiments, the lipid nanoparticles have a weight or molar ratio of lipid to nucleic acid of about 10:1 to about 100:1, e.g., about 10:1 to about 95:1, about 10:1 to about 90:1, about 10:1 to about 85:1, about 10:1 to about 80:1, about 10:1 to about 75:1, about 10:1 to about 70:1, about 10:1 to about 65:1, about 10:1 to about 60:1, about 10:1 to about 55:1, about 10:1 to about 50:1, about 10:1 to about 45:1, about 10:1 to about 40:1, about 10:1 to about 35:1, about 10:1 to about 30:1, about 10:1 to about 25:1, about 10:1 to about 20:1, about 10:1 to about 15:1.

[0160] In some embodiments, the lipid nanoparticles have an N / P ratio (i.e., the ratio of positively charged lipid amine groups to negatively charged nucleic acid phosphate groups) of about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more.

[0161] Therapeutic / preventive agents

[0162] In some embodiments, the lipid nanoparticles of the present invention also include therapeutic and / or prophylactic agents. In some embodiments, therapeutic and / or prophylactic agents described herein include organic molecules, inorganic molecules, proteins, polypeptides, nucleic acids, vaccines, immunotherapeutics, etc. In some embodiments, therapeutic and / or prophylactic agents described herein include nucleic acids. In some embodiments, therapeutic and / or prophylactic agents described herein include DNA. In some embodiments, therapeutic and / or prophylactic agents described herein include single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), genomic DNA (gDNA), complement DNA (cDNA), antisense DNA, chloroplast DNA (ctDNA or cpDNA), microsatellite DNA, mitochondrial DNA (mtDNA or mDNA), kinetoplastid DNA (kDNA), provirus, lysogen, repetitive DNA, satellite DNA, or viral DNA. In some embodiments, therapeutic and / or prophylactic agents described herein include RNA. In some embodiments, therapeutic and / or prophylactic agents described herein include small interfering RNA (siRNA). In some embodiments, therapeutic and / or prophylactic agents described herein include messenger RNA (mRNA). In some embodiments, the therapeutic and / or prophylactic agents described herein include single-stranded RNA (ssRNA), double-stranded RNA (dsRNA), precursor messenger RNA (pre-mRNA), small hairpin RNA or short hairpin RNA (shRNA), microRNA (miRNA), guide RNA (gRNA), transfer RNA (tRNA), nuclear heterogeneous RNA (hnRNA), coding RNA, non-coding RNA (ncRNA), long non-coding RNA (long ncRNA or lncRNA), satellite RNA, viral satellite RNA, signal recognition particle RNA, small cytoplasmic RNA, small nuclear RNA (snRNA), ribosomal RNA (rRNA), Piwi-interacting RNA (piRNA), polyinosinic acid, and ribozymes.

[0163] Pharmaceutical compositions and preparations

[0164] In some embodiments, the present invention provides a pharmaceutical composition comprising the lipid nanoparticles described herein and a pharmaceutically acceptable carrier. In some embodiments, the present invention provides a therapeutic and / or prophylactic agent (e.g., nucleic acid including DNA and RNA, etc.) vaccine comprising the lipid nanoparticles described herein and a pharmaceutically acceptable carrier.

[0165] In some embodiments, the pharmaceutically acceptable carriers described herein include diluents, buffers, stabilizers, and the like.

[0166] In some embodiments, the diluent comprises ethylene glycol, glycerol, polyethylene glycol, sucrose, trehalose, or a combination thereof, etc. In some embodiments, the diluent is present in an amount of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20%.

[0167] In some embodiments, the buffer comprises phosphate, citrate, imidazole, histidine, Tris, HEPES, or a combination thereof, etc. In some embodiments, the concentration of the buffer in the pharmaceutical composition is about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, or more.

[0168] In some embodiments, the stabilizer comprises a salt, including an inorganic metal salt such as sodium chloride, potassium chloride, calcium chloride, etc. In some embodiments, the concentration of the stabilizer in the pharmaceutical composition is about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM, 200 mM, or more.

[0169] In some embodiments, depending on the route of drug administration, the pharmaceutical composition and / or vaccine of the present invention can be prepared into oral preparations, intramuscular injection preparations, subcutaneous injection preparations, intravenous injection preparations, nebulized inhalation preparations, nasal spray inhalation preparations or dry powder inhalation preparations, and ophthalmic administration preparations.

[0170] Indications

[0171] The lipid nanoparticles and / or pharmaceutical compositions provided by the present invention can be used to prevent and / or treat cancer, inflammation, fibrotic diseases, autoimmune diseases, infections, mental disorders, blood diseases, chromosomal diseases, genetic diseases, connective tissue diseases, digestive diseases, ear, nose and throat diseases, endocrine diseases, eye diseases, reproductive diseases, heart diseases, kidney diseases, lung diseases, metabolic diseases, oral diseases, musculoskeletal diseases, newborn screening, nutritional diseases, parasitic diseases, skin diseases, etc.

[0172] In some embodiments, the lipid nanoparticles and / or pharmaceutical compositions provided by the present invention are mRNA vaccines, which can be used to prevent cancer, viral infections, bacterial infections, fungal infections, etc. The viruses include but are not limited to: norovirus, Ebola virus, coronavirus (including the new coronavirus SARS CoV2), cytomegalovirus, dengue virus, Zika virus, coxsackie virus, enterovirus, hepatitis virus, herpes simplex virus, human papillomavirus, influenza virus, Marburg virus, measles virus, poliovirus, rabies virus, rotavirus, measles virus, etc.

[0173] Treatment

[0174] The present invention provides a method for delivering the lipid nanoparticles described herein in vivo, comprising administering the lipid nanoparticles or pharmaceutical compositions described herein to an individual in need thereof. In some embodiments, the present invention provides a method for delivering the lipid nanoparticles described herein in vivo, comprising administering the lipid nanoparticles or pharmaceutical compositions described herein to an individual in need thereof by pulmonary delivery. In some embodiments, the present invention provides a method for delivering the lipid nanoparticles described herein in vivo, comprising administering the lipid nanoparticles or pharmaceutical compositions described herein to an individual in need thereof by intranasal delivery. In some embodiments, the present invention provides a method for delivering the lipid nanoparticles described herein in vivo, comprising administering the lipid nanoparticles or pharmaceutical compositions described herein to an individual in need thereof by inhalation. In some embodiments, the present invention provides a method for delivering the lipid nanoparticles described herein in vivo, comprising administering the lipid nanoparticles or pharmaceutical compositions described herein to an individual in need thereof by nebulized inhalation.

[0175] Preparation of lipid nanoparticles

[0176] The lipid nanoparticles of encapsulated therapeutic agent and / or preventive can be prepared using multiple methods known in the art. Typically, first prepare a solution comprising various lipid mixtures, before forming lipid nanoparticles, the solution is mixed with the solution of therapeutic agent and / or preventive, therapeutic agent and / or preventive are encapsulated in the lipid nanoparticles formed by the mixture of various lipids (such as described in WO2016004318, US20160038432). Or, first prepare a solution comprising various lipid mixtures and then form lipid nanoparticles, then the lipid nanoparticles obtained are mixed with therapeutic agent and / or preventive and therapeutic agent and / or preventive are encapsulated in the lipid nanoparticles formed by the mixture of various lipids (such as described in WO2018089801, US20180153822). These methods can effectively encapsulate therapeutic and / or prophylactic agents in lipid nanoparticles, with the encapsulation efficiency generally being not less than about 80%, not less than about 85%, not less than about 90%, not less than about 95%, not less than about 96%, not less than about 97%, not less than about 98%, and not less than about 99%.

[0177] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0178] Example

[0179] Example 1 Synthesis of Compounds CP101 / CP102

[0180] Step 1: Synthesis of 6-hydroxyhexyl 2-hexyldecanoate (Compound 3)

[0181] 2-Hexyldecanoic acid (1.84 g, 15.6 mmol) was dissolved in 200 mL of dichloromethane, and 1,6-hexanediol (2 g, 7.8 mmol) was added and stirred to dissolve. EDCI (2.24 g, 11.7 mmol) and DMAP (0.953 g, 7.8 mmol) were added and stirred at room temperature for 16 hours. Water was added to quench the mixture, and dilute hydrochloric acid was added and the mixture was separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and separated by silica gel column chromatography to obtain compound 3 (1.96 g, light yellow oil) with a yield of 70%.

[0182] 1H NMR: (400MHz, CDCl3) δ4.00(t,J=6.6Hz,2H),3.57(t,J=6.6Hz,2H),2.24(tt, J=5.2,9.0Hz,1H),1.60-1.47(m,6H),1.43(s,1H),1.37-1.30(m,8H),1.18(br s,18H),0.81(t,J=6.8Hz,6H)

[0183] Step 2: Synthesis of (Compound 6)

[0184] Compound 4 (1.77 g, 19.2 mmol), 40% aqueous NaOH solution (8.40 g, 84 mmol), and TBAB (773 mg, 0.024 mmol) were added to a flask and mixed thoroughly. Compound 5 (840 mg, 4.79 mmol) was added dropwise under ice-cooling conditions and stirred at room temperature for 15 hours. The mixture was quenched with water and separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 6 (0.88 g, light yellow oil) in a yield of 75%.

[0185] Step 3: Synthesis of (Compound 7)

[0186] Compound 6 (3 g, 13 mmol) and compound 3 (1.53 g, 4.28 mmol) were added to 10 mL of dichloromethane, and a dichloromethane solution (5 mL) of tin tetrachloride (0.338 g, 1.3 mmol) was added dropwise under an ice bath. The mixture was stirred at room temperature for 15 hours. The reaction system was added to a NaHCO3 aqueous solution, separated, and the organic phases were mixed. The organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 7 (1.51 g, colorless oil) in a yield of 60%.

[0187] 1 H NMR: (400MHz, CDCl3) δ4.90-4.71(m,1H),4.10-4.02(m,2H),3.94(br s,1H),3.60-3.43(m,10H),3.25(br d,J=6.0Hz,3H),2.67(br s,1H),2.38-2.24(m,1H),1.82-1.51(m,10H),1.49-1.18(m,30H),0.88(t,J=6.5Hz,6H)

[0188] Step 4: Synthesis of (Compound 9)

[0189] Compound 7 (2.50 g, 4.25 mmol), compound 8 (2.29 g, 5.1 mmol) and pyridine (673 mg, 8.51 mmol) were added to 20 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction solution was washed with 1N aqueous hydrochloric acid solution and brine solution, respectively. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and separated by silica gel column chromatography to obtain compound 9 (3.19 g, colorless oil) with a yield of 75%.

[0190] 1 HNMR: (400MHz, CDCl3) δ5.39(br d,J=4.3Hz,1H),5.10-4.82(m,2H),4.48(tt,J=5.4,10.8Hz,1H),4.06(t,J=6.7Hz,2H),3.64-3.36(m,8H),3.20(br s,2H),2.52-2.24(m,3H),2.06-1.70(m,7H),1.69-1.19(m,53H),1.18-1.04(m,7H),1.04-0.97(m,5H),0.96-0.83(m,15H),0.68(s,3H)

[0191] Step 5: Synthesis of compound CP101

[0192] Compound 9 (500 mg, 0.5 mmol) was added to a mixed solution of 5 mL of dichloromethane and 5 mL of 4N HCl / 1,4-dioxane, and stirred at room temperature for 3 hours. The solvent was removed by rotary evaporation, and then 10 mL of methanol was added. The pH was adjusted to neutral using ion exchange resin, and the mixture was filtered and purified by preparative liquid phase to obtain compound CP101 (360 mg, white oil) in a yield of 80%.

[0193] 1 H NMR: (400MHz, CDCl3) δ5.40(br d,J=2.0Hz,1H),4.96(t,J=5.2Hz,1H),4.56-4.42(m,1H),4.07(t,J=6.6Hz,2H),3.70-3.40(m,8H),2.80(br t,J=6.5Hz,2H),2.50-2.31(m,3H),2.12-1.80(m,5H),1.76-1.22(m,45H),1.19-0.85(m,29H),0.68(s,3H)

[0194] Step 6: Synthesis of compound CP102

[0195] Compound CP101 (900 mg, 1.0 mmol), 40% formaldehyde solution (300 mg, 4.0 mmol), and acetic acid (120 mg, 2.0 mmol) were added to 9 mL of dichloromethane and stirred at room temperature for 2 hours. NaBH(OAc)3 (636 mg, 3.0 mmol) was added and stirring was continued at room temperature for 4 hours. Saturated aqueous sodium bicarbonate solution was added to the reaction solution, extracted with dichloromethane, and the organic phases were combined and subjected to preparative liquid phase separation to obtain compound CP102 (696 mg, white oil) in a yield of 75%.

[0196] 1 H NMR: (400 MHz, CDCl3) δ 5.39 (br d,J=4.8Hz,1H),4.96(quin,J=5.2Hz,1H),4.48(tt,J=5.3,11.0Hz,1H),4 .06(t,J=6.6Hz,2H),3.67-3.34(m,8H),2.47-2.36(m,2H),2.35-2.27(m, 3H),2.22(s,5H),2.06-1.79(m,5H),1.78-1.52(m,12H),1.51-1.19(m,35 H),1.18-1.07(m,6H),1.06-0.96(m,6H),0.95-0.84(m,15H),0.68(s,3H)

[0197] The compounds of Examples 2 to 10 were prepared in a similar manner to Example 1 using corresponding starting materials.

[0198] Example 2 Synthesis of Compound CP103

[0199] 1 H NMR (400MHz, CDCl3) δ8.12-7.74(m,1H),5.35-5.21(m,1H),4.95-4.83(m,1H),4.4 7-4.22(m,1H),4.02-3.88(m,2H),3.59-3.30(m,8H),3.28-3.13(m,2H),2.52-2.3 7(m,2H),2.33-2.11(m,10H),2.00-1.64(m,8H),1.61-1.36(m,14H),1.35-1.16(m ,29H),1.12-0.99(m,7H),0.97-0.89(m,5H),0.88-0.73(m,16H),0.65-0.57(m,3H)

[0200] Example 3 Synthesis of Compound CP104

[0201] 1 H NMR (400MHz, CDCl3) δ6.67-6.42(m,1H),5.45-5.34(m,1H),5.08-4.84(m ,1H),4.55-4.35(m,1H),4.13-4.00(m,2H),3.68-3.30(m,10H),2.52-2.1 5(m,13H),2.07-1.71(m,11H),1.70-1.39(m,17H),1.36-1.21(m,25H),1. 18-1.05(m,7H),1.04-0.96(m,5H),0.94-0.83(m,15H),0.75-0.63(m,3H)

[0202] Example 4 Synthesis of Compound CP105

[0203] 1 H NMR (400MHz, CDCl3) δ7.33-7.27(m,1H),5.49-5.33(m,1H),5.05-4.87(m,1H),4.5 6-4.37(m,1H),4.13-3.98(m,2H),3.70-3.28(m,10H),3.10-2.93(m,2H),2.62-2.2 8(m,13H),2.07-1.74(m,8H),1.71-1.40(m,16H),1.40-1.32(m,7H),1.31-1.19(m ,21H),1.17-1.07(m,6H),1.05-0.95(m,6H),0.94-0.83(m,15H),0.72-0.61(m,3H)

[0204] Example 5 Synthesis of Compound CP106

[0205] 1H NMR(400MHz, CDCl3)δ8.09-7.74(m,1H),5.47-5.36(m,1H),5.07-4.89(m,1H) ,4.56-4.40(m,1H),4.11-4.03(m,2H),3.69-3.29(m,10H),2.78-2.22(m,15H) ,2.12-1.72(m,8H),1.71-1.42(m,16H),1.41-1.32(m,8H),1.32-1.21(m,21H ),1.20-1.06(m,7H),1.06-0.97(m,5H),0.97-0.78(m,16H),0.73-0.64(m,3H)

[0206] Example 6 Synthesis of Compound CP107

[0207] 1 H NMR (400MHz, CDCl3) δ6.51-6.38(m,1H),5.47-5.36(m,1H),5.11-4.88(m,1H),4.56-4.37(m,1H),4.14-4.01(m,2H),3.66 -3.40(m,8H),3.38-3.28(m,2H),2.71-2.18(m,17H),2.12-1.71(m,10H),1.70-1.43(m,15H),1.40-1.33(m,7H),1.26(br s,20H),1.20-1.07(m,7H),1.06-0.96(m,6H),0.96-0.80(m,16H),0.75-0.65(m,3H)

[0208] Example 7 Synthesis of Compound CP108

[0209] 1H NMR (400MHz, CDCl3) δ7.67(br s,1H),7.15(s,1H),7.04(s,1H),6.64-6.47(m,1H),5.38(br dd,J=4.4,13.3Hz,1H),5.09-4.95(m,1H),4.67(d,J=3.4Hz,2H),4.52-4.33(m,1H),4.07(t,J=6.7Hz ,2H),3.66-3.32(m,10H),2.45-2.25(m,3H),2.05-1.73(m,9H),1.59(dt,J=6.6,14.5Hz,9H),1.37(br d,J=3.4Hz,6H),1.26(br s,24H),1.19-1.06(m,8H),1.05-0.99(m,5H),0.94-0.85(m,17H),0.68(s,3H)

[0210] Example 8 Synthesis of Compound CP109

[0211] 1 H NMR (400MHz, CDCl3) δ8.12-7.74(m,1H),5.35-5.21(m,1H),4.95-4.83(m,1H),4.4 7-4.22(m,1H),4.02-3.88(m,2H),3.59-3.30(m,8H),3.28-3.13(m,2H),2.52-2.3 7(m,2H),2.33-2.11(m,10H),2.00-1.64(m,8H),1.61-1.36(m,14H),1.35-1.16(m ,29H),1.12-0.99(m,7H),0.97-0.89(m,5H),0.88-0.73(m,16H),0.65-0.57(m,3H)

[0212] Example 9 Synthesis of Compound CP110

[0213] 1H NMR(400MHz, CDCl3)δ5.46-5.33(m,1H),5.02(br d,J=4.8Hz,1H),4.57-4.34(m,1H),4.07(t,J=6.6Hz,2H),3.74-3.30(m,10H),2.5 0-2.26(m,5H),2.09-1.71(m,11H),1.68-1.48(m,13H),1.40-1.33(m,8H),1.26(br s,24H),1.19-1.07(m,8H),1.05-0.99(m,6H),0.95-0.83(m,18H),0.73-0.63(m,4H)

[0214] Example 10 Synthesis of Compound CP111

[0215] 1 H NMR (400MHz, CDCl3) δ6.28-6.06(m,1H),5.44-5.34(m,1H),5.10-4.86(m ,1H),4.56-4.35(m,1H),4.07(t,J=6.6Hz,2H),3.75-3.26(m,10H),3.00- 2.73(m,2H),2.46-2.24(m,5H),2.10(dd,J=2.6,7.0Hz,1H),2.05-1.94( m,4H),1.91-1.68(m,7H),1.68-1.41(m,16H),1.41-1.33(m,8H),1.26(br s,22H),1.19-1.07(m,7H),1.06-0.97(m,6H),0.95-0.84(m,16H),0.68(s,3H)

[0216] Example 11 Synthesis of Compound CP201

[0217] Step 1: Synthesis of compound 10

[0218] Compound 7 (2.0 g, 3.4 mmol) was added to a mixed solution of 20 mL of dichloromethane and 10 mL of 4N HCl / 1,4-dioxane, and stirred at room temperature for 3 hours. The solvent was removed by rotary evaporation, and then 50 mL of methanol was added to dissolve it. The pH was adjusted to neutral with ion exchange resin, and the mixture was filtered and purified by preparative liquid chromatography to obtain compound 10 (1.53, white oil) in a yield of 75%.

[0219] Step 2: Synthesis of compound 11

[0220] Compound 10 (500 mg, 1.03 mmol), compound 8 (460 mg, 1.03 mmol), pyridine (162 mg, 2.05 mmol) and DAMP (125 mg, 1.03 mmol) were added to 5 mL of dichloromethane, stirred at room temperature for 15 hours, concentrated, and separated by column chromatography to obtain compound 11 (738 mg, yellow oil) with a yield of 80%.

[0221] 1 H NMR: (400MHz, CDCl3) δ5.44-5.33(m,1H),4.95(br s,1H),4.57-4.43(m,1H),4.07(t,J=6.7Hz,2H),3.95(br s,1H),3.60-3.41(m,8H),3.35-3.22(m,2H),2.61(br s,1H),2.41-2.22(m,3H),2.05-1.74(m,7H),1.67-1.34(m,23H),1.31-1.22(m ,22H),1.19-1.08(m,6H),1.04-0.98(m,5H),0.94-0.85(m,15H),0.68(s,3H).

[0222] Step 3: Synthesis of compound CP201

[0223] Compound 11 (590 mg, 0.655 mmol), N,N-dimethylglycine (101 mg, 0.983 mmol), EDCI (151 mg, 0.786 mmol), and DMAP (96.1 mg, 0.786 mmol) were added to 5.5 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction system was poured into brine, and the organic phase was separated and purified by column chromatography to obtain compound CP201 ​​(530 mg, yellow viscous solid) in an 82% yield.

[0224] 1H NMR: (400MHz, CDCl3) δ5.42-5.33 (m, 1H), 5.22 (quin, J = 5.1Hz, 1H), 5.05 (br s,1H),4.60-4.42(m,1H),4.06(t,J=6.7Hz,2H),3.68-3.38(m,8H),3.31-3.18(m,4H),2.42-2.22(m,9H),2.06-1.92(m,2H),1.92-1 .80(m,3H),1.76(td,J=6.0,12.0Hz,2H),1.67-1.22(m,44H),1.18-1.08(m,6H),1.05-0.99(m,5H),0.93-0.84(m,15H),0.68(s,3H)

[0225] The compounds of Examples 12 to 18 were prepared in a similar manner to Example 11 using the corresponding starting materials.

[0226] Example 12 Synthesis of Compound CP202

[0227] 1 HNMR(400MHz, CDCl3)δ5.44-5.35(m,1H),5.16(quin,J=5.1Hz,1H),5.05(br s,1H),4.62-4.43(m,1H),4.07(t,J=6.7Hz,2H),3.65-3.41(m,8H),3.31-3.18(m,2H),2.67-2.59(m,2H),2.56-2.50(m,2H),2.39-2.21( m,9H),2.05-1.71(m,9H),1.66-1.46(m,16H),1.41-1.25(m,27H),1.18-1.08(m,6H),1.06-1.00(m,5H),0.95-0.84(m,15H),0.68(s,3H)

[0228] Example 13 Synthesis of Compound CP203

[0229] 1H NMR (400MHz, CDCl3) δ5.42-5.33(m,1H),5.15(quin,J=5.0Hz,2H),4.57-4.44(m,1H),4.07(t,J=6.7Hz,2H),3.63-3.40(m,8H),3.33-3.18(m,2H),2.4 3-2.19(m,13H),2.06-1.72(m,12H),1.68-1.50(m,12H),1.48-1.33(m,12H ),1.31-1.25(m,16H),1.19-0.98(m,13H),0.94-0.83(m,15H),0.68(s,3H)

[0230] Example 14 Synthesis of Compound CP204

[0231] 1 H NMR (400MHz, CDCl3) δ5.37(br d,J=4.3Hz,1H),5.19(quin,J=5.0Hz,1H),5.11(br s,1H),4.56-4.43(m,1H),4.07(t,J=6.6Hz,2H),3.66-3.37(m,9H),3.32-3.19(m,4H),2.74-2.41(m,8H),2.40-2.25(m,6H),2. 07-1.93(m,3H),1.92-1.72(m,7H),1.67-1.41(m,18H),1.40-1.26(m,22H),1.17-1.07(m,6H),1.02-0.81(m,21H),0.68(s,3H)

[0232] Example 15 Synthesis of Compound CP205

[0233] 1 H NMR (400MHz, CDCl3) δ5.44-5.35(m,1H),5.15(quin,J=5.1Hz,1H),5.07(br s,1H),4.57-4.42(m,1H),4.07(t,J=6.7Hz,2H),3.65-3.40(m,8H),3.26(br d,J=5.4Hz,2H),2.75-2.68(m,2H),2.61-2.26(m,15H),2.08-1.92(m,3H ),1.92-1.72(m,6H),1.68-1.26(m,41H),1.19-0.85(m,29H),0.68(s,3H)

[0234] Example 16 Synthesis of Compound CP206

[0235] 1 HNMR (400MHz, CDCl3) δ7.57(br s,1H),7.11(s,1H),7.00(s,1H),5.37(br d,J=1.6Hz,1H),5.23(quin,J=5.1Hz,1H),5.01-4.89(m,1H),4.79(br d,J=7.9Hz,1H),4.50(br t,J=11.3Hz,1H),4.07(t,J=6.7Hz,2H),3.66-3.34(m,8H),3.33-3.19(m,2H),2.43-2.21(m,3 H),2.08-1.93(m,3H),1.92-1.71(m,6H),1.69-1.20(m,43H),1.19-0.84(m,27H),0.68(s,3H)

[0236] Example 17 Synthesis of Compound CP207

[0237] 1 H NMR (400MHz, CDCl3) δ5.43-5.32(m,1H),5.16(quin,J=5.1Hz,1H),5.02(br s,1H),4.57-4.39(m,1H),4.06(t,J=6.7Hz,2H),3.66-3.36(m,8H),3.25(br d,J=5.8Hz,2H),2.82(br d,J=11.0Hz,2H),2.43-2.19(m,7H),2.12-1.71(m,14H),1.67-1.21(m,44H),1.18-1.07(m,6H),1.05-0.86(m,20H),0.68(s,3H)

[0238] Example 18 Synthesis of Compound CP208

[0239] 1HNMR (400MHz, CDCl3) δ5.43-5.33(m,1H),5.16(quin,J=5.1Hz,1H),5.03(br s,1H),4.56-4.42(m,1H),4.07(t,J=6.7Hz,2H),3.65-3.36(m,8H),3.25(br d,J=6.0Hz,2H),2.85(br d,J=9.9Hz,2H),2.45-2.16(m,8H),2.07-1.69(m,14H),1.68-1.21(m,46H),1.19-1.07(m,6H),1.05 0.98(m,5H),0.94-0.85(m,14H),0.68(s,3H)

[0240] Example 19 Synthesis of Compound CP301

[0241] Step 1: Synthesis of compound 12

[0242] Cholesterol (150 g, 388 mmol) was added to 1200 mL of pyridine. A solution of p-toluenesulfonyl chloride (148 g, 776 mmol) in pyridine (240 mL) was added dropwise under an ice bath and stirred overnight at room temperature. The mixture was quenched and diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to afford compound 12 (160 g, white solid) in a yield of 76.3%.

[0243] 1 H NMR: (400MHz, CDCl3) δ7.86-7.78 (m, J=8.3Hz, 2H), 7.41-7.30 (m, 2H), 5.3 5-5.30(m,1H),4.39-4.30(m,1H),2.54-2.39(m,4H),2.36-2.22(m,1H),2 .06-1.92(m,3H),1.90-1.74(m,4H),1.72-1.67(m,1H),1.59-1.31(m,11H ),1.29-1.25(m,1H),1.19-0.97(m,14H),0.94-0.87(m,10H),0.68(s,3H)

[0244] Step 2: Synthesis of compound 13

[0245] Compound 12 (50.0 g, 92.5 mmol) was added to 400 mL of 1,4-dioxane, followed by ethylene glycol (11.5 g, 185 mmol). The mixture was heated under reflux with stirring for 15 hours. The mixture was diluted with 10 times the volume of water and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 13 (23.0 g, yellow solid) in a yield of 57.8%.

[0246] 1 H NMR: (400MHz, CDCl3) δ5.38-5.33(m,1H),3.72(dd,J=3.9,5.4Hz,2H),3.63-3.55(m,2H),3.25-3 .16(m,1H),2.42-2.35(m,1H),2.27-2.16(m,1H),2.06-1.79(m,6H),1.64-1.22(m,12H),1.22(br s,2H),1.17-0.98(m,11H),0.97-0.86(m,10H),0.68(s,3H)

[0247] Step 3: Synthesis of compound 14

[0248] Compound 13 (22.5 g, 52.2 mmol) was added to 90 mL of pyridine and 90 mL of dichloromethane. DMAP (128 mg, 1.04 mmol) was then added dropwise. A solution of p-toluenesulfonyl chloride (12.0 g, 62.7 mmol) in pyridine (10 mL) was added dropwise under an ice bath. The mixture was stirred overnight at room temperature. The mixture was quenched and diluted with water, extracted with dichloromethane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to afford compound 12 (19.5 g, white solid) in a 64% yield.

[0249] 1 H NMR: (400MHz, CDCl3) δ7.87-7.82(m,J=8.1Hz,2H),7.40-7.35(m,J=8.1Hz,2H),5.35(br d,J=4.9Hz,1H),4.19(t,J=4.8Hz,2H),3.69(t,J=4.9Hz,2H),3.18-3.09(m,1H),2.48(s,3H),2.32-2 .24(m,1H),2.17-1.96(m,3H),1.92-1.79(m,3H),1.50-1.35(m,6H),1.06-0.88(m,16H),0.71(s,3H)

[0250] Step 4: Synthesis of compound 16

[0251] Compound 14 (3.00 g, 16.6 mmol) was added to 30 mL of anhydrous tetrahydrofuran, followed by NaH (998 mg, 24.9 mmol) and compound 15 (10.7 g, 18.3 mmol). The mixture was heated and stirred at reflux for 16 hours. The mixture was returned to room temperature and quenched by slowly adding ice water. The mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 16 (7.8 g, yellow solid) in a yield of 79%.

[0252] 1 H NMR: (400MHz, CDCl3)δ=7.54-7.40(m,2H),7.36-7.23(m,3H),5.55-5.45(m,1H),5 .34-5.20(m,1H),4.38-4.25(m,2H),4.09-3.94(m,2H),3.75-3.61(m,4H),3.36(br d,J=1.6Hz,1H),3.22-3.09(m,1H),2.37-2.10(m,2H),2.01-1.71(m,6H),1.52-1.22(m,12H),1.03-0.77(m,20H),0.60(s,3H)

[0253] Step 5: Synthesis of compound 17

[0254] Compound 16 (5.30 g, 8.94 mmol) was added to 53 mL of tetrahydrofuran, and hydrochloric acid (1 M, 8.94 ml) was added. After reacting at room temperature for 15 hours, 745 uL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature overnight. The pH was adjusted to neutral with saturated sodium bicarbonate aqueous solution, and ethyl acetate was added for extraction. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 17 (3.20 g, white oil) in a yield of 71%.

[0255] 1H NMR (400MHz, CDCl3) δ5.39-5.33(m,1H),3.86(br s,1H),3.96-3.56(m,1H),3.25(br s,1H),2.64(br s,2H),2.40(br d,J=10.6Hz,1H),2.30-2.17(m,1H),2.04-1.78(m,5H),1.61(br s,7H),1.25(br d,J=3.1Hz,4H),1.21-0.97(m,13H),0.94-0.83(m,10H),0.68(s,3H)

[0256] Step 6: Synthesis of compound 18

[0257] Compound 17 (2.60 g, 5.15 mmol), compound 3 (1.69 g, 5.15 mmol), EDCl (1.18 g, 6.18 mmol), and DMAP (755 mg, 6.18 mmol) were added to 20 mL of dichloromethane and stirred at room temperature for 15 hours. The mixture was quenched with water and the layers were separated. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 18 (1.9 g, yellow oil) in a yield of 46%.

[0258] 1 H NMR: (400MHz, CDCl3) δ5.38-5.30(m,1H),4.87(br s,1H),4.12(br s,2H),3.91(br s,1H),3.68(br s,3H),3.60(br d,J=8.3Hz,2H),3.22(br s,2H),2.48-2.30(m,4H),2.22(br s,1H),1.98(br d,J=18.2Hz,2H),1.50(br s,13H),1.27(br s,19H),1.17-1.04(m,7H),0.92-0.79(m,15H),0.73-0.58(m,5H)

[0259] Step 7: Synthesis of compound CP301

[0260] Compound 18 (600 mg, 736 μmol), N,N-dimethylglycine (114 mg, 1.10 mmol), EDCl (169 mg, 883 μmol), and DMAP (108 mg, 883 μmol) were added to 6 mL of dichloromethane and stirred at room temperature for 15 hours. The mixture was quenched with water and the layers were separated. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound CP301 (0.44 g, yellow oil) in a yield of 66%.

[0261] 1 H NMR: (400MHz, CDCl3) δ5.38-5.31(m,1H),4.87(quin,J=6.2Hz,1H),4.32-4.09(m,4H),3.83 (quin,J=5.2Hz,1H),3.76-3.67(m,2H),3.67-3.56(m,2H),3.26-3.10(m,3H),2.40-2.28(m, 11H),2.24-2.14(m,1H),2.06-1.93(m,2H),1.92-1.79(m,3H),1.67(td,J=3.6,7.1Hz,4H), 1.59-1.42(m,12H),1.37-1.22(m,19H),1.20-0.98(m,12H),0.95-0.83(m,16H),0.68(s,3H)

[0262] The compounds of Examples 20 to 26 were prepared in a similar manner to Example 19 using the corresponding starting materials.

[0263] Example 20 Synthesis of Compound CP302

[0264] 1H NMR(400MHz,CDCl3)δ5.34(br d,J=5.0Hz,1H),4.87(quin,J=6.3Hz,1H),4.31-4.08(m,4H),3.82(quin,J=5.1Hz,1H),3.75-3.66(m,2H) ,3.64-3.55(m,2H),3.28-3.12(m,3H),2.72-2.42(m,7H),2.40-2.12(m,10H),2.05-1.77(m,6H),1.66(br d,J=3.1Hz,4H),1.55-1.55(m,1H),1.61-1.39(m,11H),1.36-1.19(m,20H),1.17-0.98(m,11H),0.95-0.83(m,15H),0.68(s,3H)

[0265] Example 21 Synthesis of Compound CP303

[0266] 1 H NMR (400MHz, CDCl3) δ5.37-5.32(m,1H),4.87(t,J=6.3Hz,1H),4.28-4.11(m,4H),3.8 2(quin,J=5.2Hz,1H),3.76-3.71(m,2H),3.64-3.58(m,2H),3.24-3.12(m,1H),2.70-2 .46(m,4H),2.43-2.13(m,11H),2.07-1.77(m,5H),1.67(td,J=3.5,7.0Hz,5H),1.56- 1.43(m,10H),1.37-1.21(m,23H),1.18-0.98(m,12H),0.96-0.80(m,18H),0.68(s,3H)

[0267] Example 22 Synthesis of Compound CP304

[0268] 1H NMR(400MHz,CDCl3)δ5.34(br d,J=5.3Hz,1H),4.87(t,J=6.3Hz,1H),4.25-4.11(m,4H),3.81(quin,J=5.2Hz,1H ),3.76-3.69(m,2H),3.64-3.57(m,2H),3.18(tt,J=4.4,11.1Hz,1H),2.43-2.26(m ,9H),2.26-2.11(m,6H),2.07-1.75(m,8H),1.67(td,J=3.6,7.0Hz,4H),1.61-1.4 2(m,11H),1.40-1.22(m,23H),1.19-0.98(m,13H),0.95-0.82(m,17H),0.68(s,3H)

[0269] Example 23 Synthesis of Compound CP305

[0270] 1 H NMR(400MHz, CDCl3)δ5.40-5.29(m,1H),4.87(t,J=6.3Hz,1H),4.27-4.09(m,4H), 3.81(t,J=5.1Hz,1H),3.75-3.69(m,2H),3.65-3.57(m,2H),3.28-3.10(m,1H),2.7 7-2.66(m,2H),2.59-2.47(m,5H),2.41-2.24(m,9H),2.07-1.78(m,5H),1.73-1.4 3(m,17H),1.40-1.20(m,22H),1.19-0.98(m,12H),0.96-0.79(m,17H),0.69(s,3H)

[0271] Example 24 Synthesis of Compound CP306

[0272] 1H NMR (400 MHz, CDCl3) δ 5.34 (br d,J=4.9Hz,1H),4.87(t,J=6.3Hz,1H),4.26-4.08(m,3H),3.81(t,J=5. 1Hz,1H),3.75-3.70(m,2H),3.64-3.57(m,2H),3.24-3.12(m,1H),2.58- 2.25(m,16H),2.09-1.76(m,7H),1.73-1.62(m,5H),1.60-1.42(m,11H), 1.40-1.20(m,29H),1.18-0.97(m,13H),0.96-0.79(m,20H),0.68(s,3H)

[0273] Example 25 Synthesis of Compound CP307

[0274] 1 H NMR: (400MHz, CDCl3) δ5.42-5.30(m,1H),4.93-4.80(m,1H),4.24-4.06(m,4H),3.76-3.44(m,6H),3.16-3.00(m,1H),2.77-2.67(m,2H),2.53(br t,J=7.3Hz,8H),2.42-2.29(m,9H),2.22-2.12(m,1H),2.06-1.93(m,2H),1.91-1.76(m,5H),1.72-1.63(m,4H) ,1.58-1.45(m,10H),1.38-1.21(m,20H),1.18-1.06(m,6H),1.05-0.97(m,6H),0.96-0.82(m,16H),0.68(s,3H)

[0275] Example 26 Synthesis of Compound CP308

[0276] 1H NMR: (400MHz, CDCl3) δ5.34(br d,J=5.0Hz,1H),4.92-4.80(m,1H),4.25-4.05(m,4H),3.75-3.59(m,3H),3.53(br t,J=6.3Hz,2H),3.18-3.04(m,1H),2.87-2.75(m,2H),2.45-2.24(m,9H),2.23-2.12(m,1H),2.09-1.73(m,13H) ,1.72-1.59(m,5H),1.58-1.39(m,11H),1.38-1.19(m,20H),1.19-0.93(m,13H),0.93-0.82(m,15H),0.68(s,3H)

[0277] Example 27 Synthesis of Compound CP401

[0278] Step 1: Synthesis of compound 19

[0279] 1,2-Propyleneglycerol (50.0 g, 378 mmol) and triethylamine (57.4 g, 567 mmol) were dissolved and mixed in 400 mL of dichloromethane. p-Toluenesulfonyl chloride (72.1 g, 378 mmol) was dissolved in 100 mL of dichloromethane and added dropwise to the reaction system. The mixture was allowed to react overnight at room temperature. The mixture was quenched and diluted with water. The organic phases were separated and combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield compound 19 (118 g, yellow oil).

[0280] Step 2: Synthesis of compound 20

[0281] Compound 13 (10.0 g, 23.2 mmol) was added to 100 mL of anhydrous tetrahydrofuran, followed by NaH (1.86 g, 46.4 mmol). The mixture was stirred at room temperature for 2 hours, followed by compound 19 (13.3 g, 46.4 mmol). The mixture was heated and stirred at reflux for 16 hours. The reaction mixture was returned to room temperature and quenched by slowly adding ice water. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 20 (7.60 g, yellow oil) in a 60% yield.

[0282] 1H NMR: (400MHz, CDCl3) δ0.68 (s, 6H) 0.82-0.96 (m, 20H) 1.29-1.39 (m, 9H) 1.75-2.09 (m, 10H) 2.16-2.28 (m, 2H) 2.32-2.43 (m, 2H) 3.13-3.26(m,2H)3.48-3.55(m,1H)3.75(dd,J=8.29,6.50Hz,1H)4.06(dd,J=8.11,6.44Hz,1H)4.29(d,J=6.08Hz,1H)5.35(br d,J=3.70Hz,2H)

[0283] Step 3: Synthesis of compound 21

[0284] Compound 20 (7.60 g, 13.9 mmol) was added to 76 mL of tetrahydrofuran, and 3.49 mL of concentrated hydrochloric acid was added. The mixture was reacted at room temperature for 15 hours, and then extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 21 (2.60 g, yellow oil) in a yield of 40%.

[0285] 1 H NMR: (400MHz, CDCl3) δppm 0.68(s,3H)0.82-0.94(m,10H)0.97-1.21(m,12H)1.29-1.42(m,3H)1.42-1.63(m, 8H)1.77-2.05(m,5H)2.09-2.50(m,3H)3.13-3.30(m,2H)3.49-3.79(m,8H)3.88(br s,1H)5.28-5.42(m,1H)

[0286] Step 4: Synthesis of compound 22

[0287] Compound 21 (2.60 g, 5.15 mmol), compound 3 (2.03 g, 6.18 mmol), EDCl (1.18 g, 6.18 mmol), and DMAP (755 mg, 6.18 mmol) were added to 25 mL of dichloromethane and stirred at room temperature for 15 hours. The mixture was quenched with water and the layers were separated. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound 22 (2.00 g, yellow oil) in a yield of 48%.

[0288] 1H NMR: (400MHz, CDCl3) δppm 0.68 (s, 3H) 0.82-1.03 (m, 20H) 1.05-1.19 (m, 6H) 1.21-1.39 (m, 20H) 1.41-1.61 (m, 11H) 1.67 (br t,J=3.36Hz,4H)1.77-2.09(m,5H)2.23(br d,J=11.13Hz,1H)2.27-2.43(m,5H)3.14-3.27(m,1H)3.48-3.56(m,1H)3.58-3.85(m,5H)4.02(br d,J=4.16Hz,1H)4.14(dd,J=11.19,5.32Hz,2H)4.87(quin,J=6.23Hz,1H)5.35(br d,J=4.89Hz,1H)

[0289] Step 5: Synthesis of compound CP401

[0290] Compound 11 (500 mg, 613 μmol), N,N-dimethylglycine (94.8 mg, 919 μmol), EDCI (141 mg, 735 μmol), and DMAP (89.9 mg, 735 μmol) were added to 5.5 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction system was poured into brine, and the organic phase was separated and purified by column chromatography to obtain compound CP401 (0.26 g, yellow oil) in a 47% yield.

[0291] 1 H NMR: (400MHz, CDCl3) δ5.35 (br d,J=4.9Hz,1H),5.32-5.21(m,1H),4.87(quin,J=6.2Hz,1H),4.38(dd,J=3.5,12.0Hz,1H),4.17(dd,J=6.5,12.0Hz,1H),3.69-3.58(m,6H),3 .28-3.08(m,3H),2.41-2.28(m,11H),2.24-2.16(m,1H),2.04-1.95(m, 2H),1.92-1.78(m,3H),1.69-1.51(m,10H),1.46-1.38(m,3H),1.26(br s,17H),1.21-0.98(m,14H),0.96-0.84(m,19H),0.68(s,3H)

[0292] The compounds of Examples 28 to 29 were prepared in a similar manner to Example 27 using the corresponding starting materials.

[0293] Example 28 Synthesis of Compound CP402

[0294] 1 H NMR: (400MHz, CDCl3) δ5.35 (br d,J=5.0Hz,1H),5.28-5.17(m,1H),4.87(quin,J=6.2Hz,1H),4.40-4.30(m,1H), 4.23-4.13(m,1H),3.70-3.54(m,6H),3.26-3.11(m,1H),2.68-2.58(m,2H),2.54 -2.44(m,2H),2.41-2.27(m,5H),2.24(s,6H),2.05-1.94(m,2H),1.92-1.78(m,3 H),1.66(td,J=3.5,6.7Hz,4H),1.58-1.43(m,11H),1.39-1.26(m,20H),1.16(br d,J=5.3Hz,3H),1.18-1.06(m,5H),1.00(s,4H),0.94-0.84(m,17H),0.68(s,3H)

[0295] Example 29 Synthesis of Compound CP403

[0296] 1 H NMR: (400MHz, CDCl3) δ5.40-5.32(m,1H),5.27-5.18(m,1H),4.87(quin,J=6.3Hz,1H),4.4 1-4.30(m,1H),4.16(dd,J=6.4,11.9Hz,1H),3.71-3.55(m,6H),3.25-3.11(m,1H),2.41-2 .26(m,9H),2.23(s,6H),2.07-1.76(m,8H),1.66(td,J=3.6,6.7Hz,4H),1.58-1.45(m,9H) ,1.38-1.22(m,22H),1.19-1.05(m,7H),1.04-0.97(m,5H),0.94-0.83(m,16H),0.68(s,3H)

[0297] Example 30 Synthesis of Compounds CP501 / CP502

[0298] Step 1: Synthesis of compound 23

[0299] Cholesterol (40.0 g, 103 mmol) was added to 400 mL of acetone, followed by cyclopentanecarboxylic anhydride (21.3 g, 186 mmol) and triethylamine (18.8 g, 186 mmol). The mixture was reacted at 65°C for 3 days. After completion of the reaction, the solvent was removed by rotary evaporation, and the mixture was dissolved in 200 mL of dichloromethane. The mixture was washed twice with 0.5 N hydrochloric acid and then with water. The organic phase was dried over anhydrous magnesium sulfate, filtered, and concentrated. Compound 23 (45.0 g, yellow solid) was isolated by column chromatography in a yield of 86.9%.

[0300] 1 H NMR(400MHz, CDCl3)δ5.38(br d,J=4.4Hz,1H),4.76-4.54(m,1H),2.52-2.25(m,6H),2.07-1.92(m,4H),1.91-1.78(m,3H), 1.66-1.24(m,11H),1.23-0.97(m,13H),0.92(d,J=6.5Hz,3H),0.95-0.83(m,7H),0.69(s,3H)

[0301] Step 2: Synthesis of compound 24

[0302] Compound 23 (7.84 g, 15.7 mmol), compound 7 (9.20 g, 15.7 mmol), EDCI (4.50 g, 23.5 mmol), and DMAP (2.29 g, 18.8 mmol) were added to a reaction flask, followed by 92 mL of dichloromethane. The mixture was stirred at 25°C for 15 hours. After completion of the reaction, 100 mL of water was added to quench the reaction, and the mixture was extracted twice with dichloromethane. The organic phases were combined, concentrated, and separated by column chromatography to obtain compound 24 (14.0 g, yellow oil) in a yield of 83.6%.

[0303] 1 H NMR: (400MHz, CDCl3) δ5.37 (br d, J=4.1Hz, 1H), 5.14 (quin, J=5.0Hz, 1H), 4.91 (br s,1H),4.68-4.54(m,1H),4.06(t,J=6.6Hz,2H),3.64-3.35(m,8H),3.20(br d,J=5.9Hz,2H),2.46-2.23(m,7H),2.06-1.90(m,4H),1.89-1.80(m,3H),1.79-1. 70(m,2H),1.66-1.19(m,53H),1.18-0.97(m,12H),0.94-0.81(m,15H),0.68(s,3H)

[0304] Step 3: Synthesis of compound CP501

[0305] Compound 24 (14.0 g, 13.1 mmol) was added to 28 mL of dichloromethane, and 42 mL of hydrochloric acid / 1,4-dioxane was added. After stirring at room temperature for 3 hours, the solvent was removed and the system was redissolved in 100 mL of methanol. The pH value was adjusted to 7-8 with ion exchange resin. After filtration, the filter cake was washed with 100 mL of methanol, and the organic phases were combined and concentrated to obtain compound CP501 (12.7 g, yellow oil) with a yield of 100%.

[0306] 1 H NMR(400MHz, CDCl3)δ7.27-6.65(m,2H),5.37(br d,J=3.9Hz,1H),5.18(quin,J=5.1Hz,1H),4.70-4.50(m,1H),4.06(t,J=6.7Hz,2H),3.71-3.35(m,8H),3. 25-3.03(m,2H),2.53-2.21(m,7H),2.11-1.76(m,9H),1.63-1.18(m,42H),1.18-0.83(m,28H),0.67(s,3H)

[0307] Step 4: Synthesis of compound CP502

[0308] To a flask, compound CP501 (700 mg, 721 uM), N,N-dimethylglycine (112 mg, 1.08 mM), EDCI (207 mg, 1.08 mM), and DMAP (106 mg, 866 uM) were added, followed by 7 mL of dichloromethane. The mixture was stirred overnight at room temperature. After completion of the reaction, 20 mL of water was added to quench the reaction. The mixture was extracted with dichloromethane several times, and the organic phases were combined, concentrated, and separated by column chromatography to obtain compound CP502 (519 mg, yellow oil) in a yield of 68.2%.

[0309] 1 H NMR (400MHz, CDCl3) δ7.36 (br s, 1H), 5.38 (br d, J = 4.4Hz, 1H), 5.15 (br t,J=5.1Hz,1H),4.72-4.53(m,1H),4.07(t,J=6.7Hz,2H),3.64-3.29(m,10H),2.44-2 .27(m,12H),2.05-1.76(m,11H),1.68-1.22(m,44H),1.19-0.85(m,28H),0.69(s,3H)

[0310] The compounds of Examples 31 to 38 were prepared in a similar manner to Example 30 using the corresponding starting materials.

[0311] Example 31 Synthesis of Compound CP503

[0312] 1 H NMR (400MHz, CDCl3) δ8.01 (br d, J=1.1Hz, 1H), 5.38 (br d,J=4.5Hz,1H),5.17(quin,J=5.2Hz,1H),4.67-4.56(m,1H),4.07(t,J=6.7Hz,2H),3.64-3.38(m,8H),3.31(q,J=6.5Hz,2H),2.58(br t,J=6.0Hz,2H),2.45-2.25(m,15H),2.06-1.72(m,11H),1.66-1.44(m,15H) ),1.39-1.23(m,28H),1.19-1.00(m,12H),0.94-0.85(m,14H),0.69(s,3H)

[0313] Example 32 Synthesis of Compound CP504

[0314] 1 H NMR(400MHz,CDCl3)δ6.57(br s,1H),5.38(br d,J=4.4Hz,1H),5.19(quin,J=5.1Hz,1H),4.70-4.53(m,1H),4.07(t,J=6.7Hz,2H),3.61-3.29(m, 10H),2.47-2.20(m,16H),2.08-1.71(m,12H),1.68-1.19(m,44H),1.19-0.85(m,27H),0.68(s,3H)

[0315] Example 33 Synthesis of Compound CP505

[0316] 1H NMR(400MHz, CDCl3)δ5.38(br d,J=4.3Hz,1H),5.17(quin,J=5.1Hz,1H),4.71-4.54(m,1H),4.07(t,J=6.6Hz,2H),3.64-3.28(m,10H), 3.01(s,2H),2.70-2.23(m,17H),2.05-1.76(m,10H),1.67-1.22(m,44H),1.18-0.83(m,28H),0.69(s,3H)

[0317] Example 34 Synthesis of Compound CP506

[0318] 1 H NMR(400MHz, CDCl3)δ8.05-7.73(m,1H),5.38(br d,J=4.1Hz,1H),5.17(t,J=5.2Hz,1H),4.70-4.53(m,1H),4.07(t,J=6.6Hz,2H),3.64-3.23(m,1 0H),2.68-2.28(m,20H),2.04-1.74(m,10H),1.67-1.20(m,44H),1.18-0.85(m,28H),0.69(s,3H)

[0319] Example 35 Synthesis of Compound CP507

[0320] 1H NMR(400MHz,CDCl3)δ6.44(br s,1H),5.38(br d,J=4.0Hz,1H),5.20(quin,J=5.1Hz,1H),4.68-4.53(m,1H),4.07(t,J=6.7Hz,2H),3.6 6-3.21(m,10H),2.68-2.16(m,21H),2.06-1.70(m,12H),1.69-0.80(m,71H),0.68(s,3H)

[0321] Example 36 Synthesis of Compound CP508

[0322] 1H NMR (400MHz, CDCl3) δ7.64(s,1H),7.14(s,1H),7.04(s,1H),6.41(br s,1H),5.38(br d,J=4.1Hz,1H),5.26-5.14(m,1H),4.75-4.52(m,3H),4.07(t,J=6.7Hz,2H),3.61-3.24(m,10H),2.49-2. 23(m,7H),2.06-1.91(m,7H),1.90-1.81(m,3H),1.73(td,J=5.5,11.4Hz,2H),1.67-1.32(m,23H),1.26(br s,20H),1.19-1.08(m,6H),1.06-0.98(m,5H),0.95-0.84(m,14H),0.68(s,3H)

[0323] Example 37 Synthesis of Compound CP509

[0324] 1 H NMR(400MHz,CDCl3)δ6.16(br s,1H),5.37(br d,J=4.1Hz,1H),5.20(quin,J=5.2Hz,1H),4.69-4.54(m,1H),4.07(t,J=6.7Hz,2H),3.64-3.20(m,10H),2.90(br d,J=10.8Hz,2H),2.46-2.21(m,10H),2.13-1.71(m,17H),1.67-0.83(m,70H),0.68(s,3H)

[0325] Example 38 Synthesis of Compound CP510

[0326] 1H NMR(400MHz,CDCl3)δ6.14(br s,1H),5.38(br d,J=4.1Hz,1H),5.20(quin,J=5.2Hz,1H),4.69-4.52(m,1H),4.07(t,J=6.6Hz,2H),3.65-3.23(m, 10H),3.00-2.72(m,2H),2.47-2.25(m,9H),2.10(d,J=7.0Hz,2H),2.05-0.81(m,88H),0.68(s,3H)

[0327] Example 39: Preparation and Characterization of mRNA-LNP

[0328] 1. Materials and Instruments

[0329] Table 1 Main experimental consumables

[0330] Table 2 Main experimental equipment

[0331] Table 3 Other main reagents

[0332] 2. Experimental Plan

[0333] Preparation of mRNA-LNPs

[0334] The lipid compound of the present invention, phospholipid (DOPE), and PEG lipid (DMG-PEG) were mixed in an ethanol solution at a molar ratio of 49.25:49.25:1.5, and the mRNA was diluted to a final concentration of 135 ng / uL in 25 mM sodium acetate buffer at pH 4.0. The aqueous and ethanolic phases were mixed using a Precision Nanosystems microfluidic device with a mixing flow rate of 9 mL / min for the aqueous phase and 3 mL / min for the ethanolic phase. The prepared encapsulation solution was diluted 40-fold into a pH 7.5 25 mM Tris 25 mM sodium acetate buffer solution. After ultrafiltration using a 30 kDa ultrafiltration tube, 25% of the final volume of 435 mg / mL sucrose in 20 mM Tris 10.7 mM sodium acetate buffer solution was added. After sterile filtration, the mRNA-LNP experimental sample was obtained.

[0335] Characterization of mRNA-LNPs

[0336] The prepared mRNA-LNP experimental sample was diluted 50 times with buffer (final concentration was 2-100 ng / μL), and the average particle size, PDI and ζ potential of the nanoparticles were measured using a Malvern particle size analyzer; the average particle size and PDI were measured using a ZEN0040 DLS sample cell with a sample volume of 200 μL; the ζ potential was measured using a DTS1070 potential cell with a sample volume of 800 μL. The mRNA content and encapsulation efficiency were measured using a Quant-iT TM RiboGreen RNA detection kit, TE buffer to detect free mRNA content C 游离 , 2% Triton buffer was used to detect the total mRNA content C 总 The encapsulation efficiency is calculated by the formula EE=(1-C 游离 / C 总 The experimental results are shown in Table 4.

[0337] Table 4 Physicochemical characterization of GFPmRNA-LNP *Note: For comparison with lipid compounds, the synthesis of each compound was based on the method provided in the corresponding patent examples, and structural characterization showed that the compound structure was correctly synthesized.

[0338] The results show that the mRNA-LNP formed by the cationic lipid compound of the present invention and phospholipids (DOPE), PEG lipids (DMG-PEG) and mRNA has good physicochemical parameters, with an average particle size in the range of about 60-105 nm, a PDI of less than 0.2, a good polydispersity coefficient, a zeta potential between -15 mV and 15 mV, and an LNP encapsulation efficiency of mRNA greater than 80%.

[0339] Example 40 mRNA-LNP in vitro cell transfection activity

[0340] Fluorescence microscopy was used to detect the expression level of green fluorescent protein (eGFP) to evaluate the transfection activity of mRNA-LNP on HEK293T cells. 5 A HEK293T cell solution of 1 mL / well was inoculated into a 24-well cell culture plate. After 24 hours, each well was transfected with 500 ng of eGFP mRNA-LNP, and the cell culture plate was placed in a 37°C, 5% CO2 cell culture incubator. The negative control group was transfected with an equal volume of normal saline. After 24 hours, microscopic imaging was performed, and the results are shown in Figure 1. The results show that the three-component LNP composition formed by the cationic lipid compound of the present invention can achieve high expression of eGFP-mRNA in cells, and the expression level is better than that of the control group, and the expression level is better than that of other compounds disclosed in the patent (compound (3), CLinDMA, HGT4001, ICE).

[0341] Example 41 mRNA-LNP Animal Immunization Test

[0342] We used SARS-CoV-2 S protein mRNA to evaluate the immunogenicity of mRNA-LNPs in mice. The LNP formulation used was an ionizable cationic lipid compound: DOPE:DMG-PEG2K at a molar ratio of 49.25:49.25:1.5 for mRNA encapsulation. See Table 4 for the specific formulation. Six- to eight-week-old female BALB / c mice were randomly divided into groups of six and immunized via intramuscular injection into the hind leg. Immunizations were administered on days 0 and 14, with a dose of 5 μg of mRNA-LNP. On day 28, blood was collected and serum was separated. Antibody titers specific for the SARS-CoV-2 S protein antigen were assessed by ELISA. PBMCs were then collected and assayed for S protein-specific IFNγ-ELISPOT. The antibody titer values ​​(GMTs) (95% CIs) are shown in Figure 2. These results demonstrate that the mRNA vaccine composition formed from lipid nanoparticles provided by the present invention exhibits higher immunogenicity than the control group. The ELISPOT data, shown in Figure 3, demonstrate that the mRNA vaccine composition formed from lipid nanoparticles provided by the present invention can induce higher cellular immunity.

[0343] Example 42 mRNA-LNP Safety Evaluation

[0344] The CCK-8 method was used to evaluate the effect of mRNA-LNP on the growth status of HEK293T cells. HEK293T cells were plated in 96-well plates and 10 cells were seeded per well. 4 Cells were transfected with 2 μg of mRNA-LNP 24 hours later (transfection volume 20 μL, culture medium volume 10%, final concentration of cationic lipid compound approximately 180 μM). 10% DMSO was selected as a positive control, and PBS was selected as a negative control. Three wells were cultured in parallel at 37°C, 5% CO2 for another 24 hours. After adding CCK-8 substrate and incubating for 2 hours, the absorbance was measured by a microplate reader, and the relative cell survival rate was calculated. The experimental results are shown in Figure 4, which show that the compounds provided by the present invention had no effect on cell proliferation and had good safety.

[0345] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A lipid compound represented by formula (I): or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein, L1, L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G1, G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; One of R1, R2 and R3 is selected from an optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein the C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by -O-, -S-, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl substitution; At the same time, another one of R1, R2 and R3 is selected from a steroidal group; At the same time, the third one of R1, R2 and R3 is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxyl, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy; R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkyne; R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl; m, n and p are each independently selected from 1, 2 or 3; q is selected from 0 or 1.

2. The compound according to claim 1 or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L1 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G1 is independently selected at each occurrence from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; R1 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, -NR a -, carbocyclyl, aryl, heteroaryl, and / or heterocyclyl substitution; L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; One of R2 and R3 is selected from a steroidal group, while the other is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxyl, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy; R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkyne; R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl; m, n and p are each independently selected from 1, 2 or 3; q is selected from 0 or 1.

3. The compound according to claim 2 or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G2 is independently selected from -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; R2 is selected from a steroid group; L3 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G3 is independently selected at each occurrence from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; R3 is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q -guanidine; wherein The nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidinyl group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxyl, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy; or L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G2 is independently selected at each occurrence from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; R2 is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxyl, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy; L3 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G3 is independently selected from -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; R3 is selected from a steroid group; R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkyne; R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl; m, n and p are each independently selected from 1, 2 or 3; q is selected from 0 or 1.

4. The compound according to claim 1 or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L2 is independently selected at each occurrence from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G2 is independently selected at each occurrence from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; R2 is selected from optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl; wherein C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 One or more -CH2- in the alkynyl group may be optionally replaced by O, S, carbocyclyl, aryl, heteroaryl, heterocyclyl; L1 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Alkenylene, optionally substituted C2-C 20 Alkyne, optionally substituted C1-C 20 acyl group; G1 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; One of R1 and R3 is selected from a steroidal compound group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -nitrogen-containing heteroaryl, -(R4) q -nitrogen-containing heterocyclic group, -(R4) q wherein the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidine group are optionally substituted by one or more groups selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxy, halogen, hydroxyl, mercapto, cyano, nitro, amino, carboxyl, C1-C 20 Acyl, C1-C 20 acyloxy; R4 is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkyne; R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 Alkynyl, optionally substituted C3-C 14 Carbocyclic group, optionally substituted C6-C 14 aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl; m, n and p are each independently selected from 1, 2 or 3; q is selected from 0 or 1.

5. A compound according to any one of the preceding claims, selected from or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein, L1, L2, L3, G1, G2, G3, R1, R2 and R3 are as defined in any one of claims 1-4.

6. A compound according to any one of the preceding claims, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; in, The steroidal compound in the steroidal compound group is selected from naturally occurring steroidal compounds or their analogs; preferably, it includes plant sterols and animal sterols, or their analogs; more preferably, it is selected from cholesterol and its derivatives.

7. A compound according to any one of the preceding claims, or a stereoisomer, a tautomer, and a pharmaceutically acceptable salt thereof; in, The steroidal compound group has the following structure: R5 is selected from hydrogen, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C1-C 20 Alkoxycarbonyl C1-C 20 alkyl-; R6 is selected from hydrogen, halogen, cyano, hydroxyl, amino, oxo, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl; m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; Preferably, the steroid group is selected from: Where R' is C 1-20 alkyl.

8. A compound according to any one of the preceding claims, which is a compound represented by formula (IV-a) or formula (IV-b): or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof, wherein L1, L2, L3, G1, G2, G3, R1, R2, R3, m, n, p are as defined in any one of the preceding claims.

9. A compound according to any one of the preceding claims, which is a compound represented by formula (Va) or formula (Vb): or its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein L1, L2, L3, G1, G2, G3, R1, R2, R3 are as defined in any of the preceding claims.

10. The compound according to any one of the preceding claims, which is a compound represented by formula (VI-a) or formula (VI-b): or its stereoisomers, tautomers, and pharmaceutically acceptable salts, wherein L1, L2, L3, G1, G2, G3, R1, R2, R3 are as defined in any of the preceding claims.

11. A compound according to any one of the preceding claims, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; wherein Each occurrence of L1 is independently selected from optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 Acyl; preferably, L1 is selected from optionally substituted C1-C6 alkylene, optionally substituted C2-C6 acyl; G1 is independently selected at each occurrence from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; preferably, G1 is selected from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-、-N(R a )C(=O)O-、-OC(=O)N(R a )-; More preferably, G1 is selected from -C(=O)O-, -OC(=O)-; R a and R b are each independently selected from H, optionally substituted C1-C 20 Alkyl, optionally substituted C2-C 20 Alkenyl, optionally substituted C2-C 20 alkynyl, optionally substituted carbocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl; preferably, R a and R b Each independently selected from H, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 More preferably, R a and R b Each independently selected from H, C1-C 20 alkyl; R1 is selected from optionally substituted C1-C 20 Alkyl; wherein C1-C 20 One or more -CH2- in the alkyl group may be optionally replaced by O, S, -NR a -, carbocyclic group substitution.

12. A compound according to any one of the preceding claims, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; wherein L1 is independently selected at each occurrence from optionally substituted methylene, ethylene, propylene, butylene, pentylene, hexylene, acetyl, propionyl, butyryl, pentanoyl, hexanoyl; G1 is independently selected at each occurrence from -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)N(R a )-、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)N(R a )-, -SS-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; R a and R b Each independently selected from H, C1-C6 alkyl; R1 is selected from 13. A compound according to any one of the preceding claims, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; wherein L2 and L3 are each independently selected from a bond, an optionally substituted C1-C 20 Alkylene, optionally substituted C2-C 20 acyl group; G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; One of R2 and R3 is selected from a cholesterol group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -5 or 6-membered nitrogen-containing heteroaryl, -(R4) q -5 or 6-membered nitrogen-containing heterocyclic group; wherein The 5- or 6-membered nitrogen-containing heteroaryl and the 5- or 6-membered nitrogen-containing heterocyclic group are optionally substituted by a group selected from the following: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogen, hydroxyl, thiol, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy; R4 is selected from C1-C6 alkylene; R a and R b Each independently selected from H, C1-C6 alkyl; q is selected from 0 or 1.

14. A compound according to any one of the preceding claims, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; wherein L2 and L3 are each independently selected at each occurrence from a bond, an optionally substituted C1-C6 alkylene, an optionally substituted C2-C6 acyl; G2 and G3 are each independently selected from a bond, -O-, -S-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -C(=O)NR a -、-NR a C(=O)-, -S(=O)O-, -OS(=O)O-, -S(=O)2O-, -OS(=O)2O-, -C(=O)S-, -C(=S)S-, -OP(=O)(OR a )O-、-N(R a )C(=O)O-、-OC(=O)NR a -, -SS-, -OC(=O)S-, -SC(=O)O-, -NR a C(=O)NR b -; One of R2 and R3 is selected from a cholesterol group, and the other is selected from -(R4) q -NR a R b 、-(R4) q -5 or 6-membered nitrogen-containing heteroaryl, -(R4) q -5 or 6-membered nitrogen-containing heterocyclic group; wherein The 5- or 6-membered nitrogen-containing heteroaryl and the 5- or 6-membered nitrogen-containing heterocyclic group are optionally substituted by a group selected from the following: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogen, hydroxyl, thiol, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy; R4 is selected from C1-C6 alkylene; R a and R b Each independently selected from H, C1-C6 alkyl; q is selected from 0 or 1.

15. The compound according to claim 1, selected from: or its stereoisomers, tautomers, and pharmaceutically acceptable salts.

16. A lipid nanoparticle comprising the lipid compound according to any one of claims 1 to 15 or its stereoisomers, tautomers, and pharmaceutically acceptable salts thereof.

17. The lipid nanoparticle of claim 16, further comprising phospholipids and / or polyethylene glycol lipids.

18. The lipid nanoparticle of claim 17, further comprising a therapeutic and / or preventive agent.

19. The lipid nanoparticle according to claim 18, wherein the therapeutic and / or preventive agent comprises one or more nucleic acids, such as DNA, RNA, etc.

20. A pharmaceutical composition comprising the lipid nanoparticle according to any one of claims 16-19 and a pharmaceutically acceptable carrier.

21. A method for treating and / or preventing a disease, comprising administering a therapeutically effective amount of the lipid nanoparticle according to any one of claims 16-19 or the pharmaceutical composition according to claim 20 to an individual in need thereof.

22. Use of a compound according to any one of claims 1 to 15 and / or a lipid nanoparticle according to claims 16 to 19 in the preparation of a therapeutic and / or preventive agent delivery system.

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