Ionizable cationic lipid compound and use thereof

Abstract

The present invention provides an ionizable cationic lipid compound having a structure represented by formula (I), which can be used for preparing a lipid nanoparticle (LNP) for the delivery of therapeutic and / or prophylactic agents. The LNP prepared by using the ionizable cationic lipid compound of the present invention has better stability and transfection efficiency, and can efficiently and stably deliver a bioactive substance (comprising nucleic acids, such as mRNA) to a target cell or organ, thereby causing 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] Ar is an aryl or heteroaryl group; the aryl or heteroaryl group is optionally substituted by a group selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, carbocyclyl, aryl, halogen, alkoxy, alkylthio, NR4R4', R4-C(O)-, R4-C(O)O-;

[0008] G1, G2 and G3 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 alkynylene, optionally substituted carbocyclylene, optionally substituted arylene;

[0009] L1, L2, L3, L4, L5 and L6 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 )-;

[0010] 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;

[0011] At the same time, another one of R1, R2 and R3 is selected from a steroid group;

[0012] 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, R c O-C1-C 20 alkyl;

[0013] R4 and R4' are each independently selected from H, Cl-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, carbocyclyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl and / or heterocyclylalkyl;

[0014] R4" is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0015] R a 、R b and R c 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 arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl;

[0016] m, n and p are each independently selected from 1, 2 or 3;

[0017] q is selected from 0 or 1.

[0018] 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.

[0019] In another aspect, the present invention also provides a pharmaceutical composition comprising the lipid nanoparticles described herein and a pharmaceutically acceptable carrier.

[0020] 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.

[0021] 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:

[0022] 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

[0023] FIG1 shows the transfection efficiency of Luc-mRNA-LNP in HEK293T cells detected by fluorescence microscopy.

[0024] Figure 2 shows the serum-specific antibody titer after mice were immunized with the SARS-CoV-2 S protein mRNA-LNP vaccine.

[0025] Figure 3 shows the level of specific CD8+ T cell response in the spleen of mice after immunization with SARS-CoV-2 S protein mRNA-LNP.

[0026] Figure 4 shows the specific CD4+ T cell response level in the spleen of mice after immunization with SARS-CoV-2 S protein mRNA-LNP. DETAILED DESCRIPTION

[0027] definition

[0028] 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.

[0029] 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.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] 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.

[0034] 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.

[0035] 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.

[0036] 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.

[0037] 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.

[0038] 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.

[0039] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0040] 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.

[0041] 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.

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] As used herein, the term "steroid" is an organic compound having a four-ring carbon skeleton structure as shown below.

[0049] 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.

[0050] 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.

[0051] 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.

[0052] 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] Compound

[0057] In some embodiments, the present invention provides a lipid compound represented by formula (I)

[0058] or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein L1, L2, L3, L4, L5, L6, G1, G2, G3, R1, R2, R3, m, n, and p are as defined above.

[0059] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0060] Ar is an aryl group; the aryl group is optionally substituted by a group selected from the following: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, carbocyclyl, aryl, halogen, alkoxy, alkylthio, NR4R4', R4-C(O)-, R4-C(O)O-;

[0061] G1, G2 and G3 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 alkynylene, optionally substituted carbocyclylene, optionally substituted arylene;

[0062] L1, L2, L3, L4, L5 and L6 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 )-;

[0063] 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;

[0064] R2 is selected from a steroid group;

[0065] 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, R c O-C1-C 20 alkyl;

[0066] R4 and R4' are each independently selected from H, Cl-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, carbocyclyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl and / or heterocyclylalkyl;

[0067] R4" is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0068] R a 、R b and R c 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 arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl;

[0069] m, n and p are each independently selected from 1, 2 or 3;

[0070] q is selected from 0 or 1.

[0071] In some embodiments, the present invention provides a lipid compound represented by formula (I) or its stereoisomers, tautomers, and pharmaceutically acceptable salts; wherein,

[0072] Ar is phenyl; said phenyl is optionally substituted by a group selected from the group consisting of: C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, carbocyclyl, aryl, halogen, alkoxy, alkylthio, NR4R4', R4-C(O)-, R4-C(O)O-;

[0073] G1, G2 and G3 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;

[0074] L1, L2, L3, L4, L5 and L6 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 )-;

[0075] 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;

[0076] R2 is selected from a steroid group;

[0077] R3 is selected from -(R4") q -NR a R b 、-(R4”) q -nitrogen-containing heteroaryl, -(R4") q- nitrogen-containing heterocyclic group; wherein the nitrogen-containing heteroaryl group and the nitrogen-containing heterocyclic 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, R c O-C1-C 20 alkyl;

[0078] R4 and R4' are each independently selected from H, Cl-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, carbocyclyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl and / or heterocyclylalkyl;

[0079] R4" is selected from C1-C 20 Alkylene, C2-C 20 Alkenylene, C2-C 20 Alkynylidene;

[0080] R a 、R b and R c 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 arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl;

[0081] m, n and p are each independently selected from 1, 2 or 3;

[0082] q is selected from 0 or 1.

[0083] In some embodiments, the present invention provides lipid compounds represented by formula (II-1), (II-2), (II-3), (II-4), and (II-5):

[0084] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof; wherein L1, L2, L3, L4, L5, L6, G1, G2, G3, R1, R2, R3, m, n and p are as defined herein.

[0085] In some embodiments, the present invention provides lipid compounds represented by formula (III-1), (III-2), (III-3), (III-4), and (III-5):

[0086] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof; wherein L1, L2, L3, L4, L5, L6, G1, G2, G3, R1, R2, R3, m, n and p are as defined herein.

[0087] In some embodiments, the present invention provides lipid compounds represented by formula (IV-1), (IV-2), (IV-3), (IV-4), (IV-5), (IV-6), (IV-7), (IV-8), (IV-9), and (IV-10):

[0088] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof; wherein R1, R2, and R3 are as defined herein, and r, s, t, and u are each independently selected from an integer of 1-10.

[0089] 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.

[0090] 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.

[0091] 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:

[0092] 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-;

[0093] R6 is selected from hydrogen, halogen, cyano, hydroxy, amino, oxo, C1-C 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl;

[0094] m is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0095] In some embodiments, the steroid group is selected from:

[0096] Where R' is C 1- C 20 alkyl.

[0097] In some embodiments, the present invention provides a lipid compound represented by formula (V-1), (V-2), (V-3), (V-4), (V-5), (V-6), (V-7), (V-8), (V-9), or (V-10):

[0098] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof; wherein R1, R3, r, s, t and u are as defined herein.

[0099] In some embodiments, the present invention provides a lipid compound represented by formula (VI-1), (VI-2), (VI-3), (VI-4), (VI-5), (VI-6), (VI-7), (VI-8), (VI-9), or (VI-10):

[0100] or stereoisomers, tautomers, and pharmaceutically acceptable salts thereof; wherein R1, R3, r, s, t and u are as defined herein.

[0101] In some specific embodiments, R1 is selected from optionally substituted C1-C 20 Alkyl; wherein the C1-C 20 One or more -CH2- in the alkyl group may be optionally replaced by O, S, -NR a -, carbocyclic group substitution.

[0102] In some specific embodiments, R1 is selected from

[0103] In some specific embodiments,

[0104] R3 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, R c O-C1-C6 alkyl;

[0105] R4" is selected from C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene;

[0106] R a and R b Each independently selected from H and C1-C6 alkyl;

[0107] R c Selected from H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, phenyl-C1-C6 alkyl;

[0108] q is selected from 0 or 1.

[0109] In some specific embodiments,

[0110] R3 is selected from -R4"-NR a R b, -R4"-5 or 6-membered nitrogen-containing heterocyclic group; wherein the 5 or 6-membered nitrogen-containing heterocyclic group is 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, R c O-C1-C6 alkyl;

[0111] R4" is selected from C1-C6 alkylene;

[0112] R a and R b Each independently selected from H, C1-C6 alkyl;

[0113] R c Selected from H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, benzyl.

[0114] In some embodiments, the present invention provides a compound selected from the following or a pharmaceutically acceptable salt thereof.

[0115] Lipid nanoparticles (LNPs)

[0116] 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)).

[0117] 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 40-80:20-50:0.5-10; more preferably, the molar ratio of lipid compound: phospholipid: PEG-lipid is 50-70:30-40:0.5-5; most preferably, the molar ratio of lipid compound: phospholipid: PEG-lipid is 65.7:32.8:1.5.

[0118] phospholipids

[0119] 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.

[0120] 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%.

[0121] PEG lipids

[0122] 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.

[0123] 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.

[0124] 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).

[0125] 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%.

[0126] Particle size of lipid nanoparticles

[0127] 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.

[0128] Lipid / nucleic acid ratio

[0129] 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.

[0130] 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.

[0131] Therapeutic / preventive agents

[0132] 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.

[0133] Pharmaceutical compositions and preparations

[0134] 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.

[0135] In some embodiments, the pharmaceutically acceptable carriers described herein include diluents, buffers, stabilizers, and the like.

[0136] 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%.

[0137] 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.

[0138] 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.

[0139] 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.

[0140] Indications

[0141] 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.

[0142] 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, etc.

[0143] Treatment

[0144] 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.

[0145] Preparation of lipid nanoparticles

[0146] 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%.

[0147] 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.

[0148] Example

[0149] Example 1 Synthesis of Compound 1

[0150] Step 1: Synthesis of Compound A2

[0151] 10 g of cholesterol (A1) was dissolved in dichloromethane (150 mL), and 4-bromobutyric acid (4.75 g) and DMAP (950 mg) were added thereto. After the mixture was dissolved, the mixture was stirred in an ice bath, EDCI (g) was added, and the mixture was reacted at room temperature for 16 hours. After the reaction was completed, 100 mL of water was added to the system, the organic phase was separated, and the aqueous phase was extracted with dichloromethane (50 mLх2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by column chromatography to obtain compound A2 (10 g, white solid) in a yield of 72%.

[0152] Step 2: Synthesis of Compound A4

[0153] Compound A3 (30 g) and DMF (0.1 mL) were added to a 250 mL round-bottom flask. Dissolve and mix in 100 mL of dichloromethane. Then, 49.5 mL of oxalyl chloride was added dropwise to the reaction flask. The mixture was allowed to react at room temperature for 12 hours. After the reaction, the solvent was removed by rotary evaporation to obtain compound A4 (32.1 g, yellow oil) in a 100% yield.

[0154] Step 3: Synthesis of Compound A5

[0155] To a 500 mL round-bottom flask, 6-bromo-1-hexanol (25.4 g) and dichloromethane (100 mL) were added separately. 32.1 g of Compound A4 was dissolved in 100 mL of dichloromethane and slowly added dropwise to the reaction flask. The reaction was continued at room temperature for 12 hours. After the reaction, the solvent was directly removed by rotary evaporation, and Compound A5 (39.5 g, light yellow oil) was isolated by silica gel column chromatography in an 80% yield.

[0156] Step 4: Synthesis of Compound A7

[0157] Compound A6 (10 g) and DMF (100 mL) were added to a 250 mL round-bottom flask. Imidazole (6.32 g) and tert-butyldimethylsilyl chloride (11.3 g) were added under ice bath conditions and stirred at room temperature for 16 hours. After the reaction was completed, the reaction system was diluted with 500 mL 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 silica gel column chromatography to obtain compound A7 (14.0 g, bright yellow oil) in a yield of 77%.

[0158] Step 5: Synthesis of Compound A8

[0159] To a 500 mL round-bottom flask were added compound A7 (7.28 g), compound A5 (12.0 g), DMF (100 mL), potassium carbonate (19.7 g), and NaI (770 mg), respectively. The reaction system was stirred at 60°C for 16 hours. After completion of the reaction, the reaction system was poured into 500 mL of water and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound A8 (11.5 g, bright yellow oil) in a yield of 38%.

[0160] Step 6: Synthesis of Compound A9

[0161] Compound A8 (7.60 g, 12.8 mmol), compound A2 (7.55 g, 14.1 mmol), potassium carbonate (5.31 g, 38.4 mmol), and sodium iodide (190 mg, 0.149 mmol) were dissolved in 75 mL of DMF and stirred at 60°C for 16 hours. 400 mL of water was added to the reaction system to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound A9 (9.50 g, bright yellow oil) in a 70% yield.

[0162] Step 7: Synthesis of Compound A10

[0163] Compound A9 (5.00 g, 4.77 mmol) was dissolved in 50 mL of tetrahydrofuran, and then 1.0 M TBAF solution (5.8 mL) was added. The mixture was stirred at room temperature for 2 hours, and then 200 mL of water was added to quench the reaction. The reaction system was extracted with ethyl acetate (100 mLх3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound A10 (4.20 g, yellow oil) in a yield of 94%.

[0164] 1 H NMR (400MHz, CDCl3): δ6.51 (s, 2H), 6.37 (s, 1H), 5.38 (m, 1H), 5.29 (d, J = 17.2Hz, 1H), 4.63 (m, 3H) ,4.09(t,J=6.4Hz,2H),4.00(t,J=6.4Hz,2H),3.94(t,J=6.4Hz,2H),2.49(t,J=7.2Hz,2H),2.32(d ,J=8.0Hz,3H),2.13-2.08(m,6H),2.05-1.93(m,3H),1.91-1.74(m,6H),1.71-1.54(m,8H),1.53-1 .39(m,11H),1.37-1.21(m,24H),1.14-1.09(m,7H),1.02(s,3H),0.96-0.84(m,13H),0.69(s,3H).

[0165] Step 8: Synthesis of Compound A11

[0166] Compound A10 (1.50 g, 1.60 mmol) and pyridine (410 mg, 3.21 mmol) were dissolved in 15 mL of dichloromethane, followed by the addition of SOCl2 (290 mg, 2.41 mmol) and stirring at room temperature for 2 hours. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to the reaction system until the pH reached 8. The aqueous phase was extracted twice with dichloromethane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to afford compound A11 (1.50 g, yellow oil) in a 98% yield.

[0167] Step 9: Synthesis of Compound 1

[0168] Compound A11 (1.50 g, 1.57 mmol), 1-(3-aminopropyl)imidazole (990 mg, 7.88 mmol), potassium carbonate (440 mg, 3.15 mmol), potassium iodide (50 mg, 0.105 mmol), and 15 mL of DMF were added to a reaction flask. After the mixture dissolved, it was heated to 60° C. and reacted for 16 hours. After the reaction was completed, 20 mL of water was added to the reaction system, and the aqueous phase was extracted with ethyl acetate (20 mLх3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 1 (450 mg, yellow oil) in a yield of 27%.

[0169] 1 HNMR(400MHz, CDCl3)δ7.48(s,1H),7.05(s,1H),6.91(s,1H),6.45(d,2H),6.35(s,1H),5.37(m,1H ),4.64(m,1H),4.08(m,4H),3.99(m,2H),3.93(m,2H),3.70(m,2H),2.62(m,2H),2.49(m,2H),2.33( m,3H),2.10(m,2H),2.04-1.93(m,4H),1.90-1.74(m,6H),1.64(s,5H),1.58-1.53(m,4H),1.44(m, 10H),1.30(m,24H),1.13(m,7H),1.02(m,4H),0.96(m,2H),0.92(s,3H),0.88(m,10H),0.68(s,3H).

[0170] LC-MS(ESI+):1041

[0171] Example 2 Synthesis of Compound 2

[0172] Compound A10 (560 mg, 600 μmol), N,N-dimethylglycine (92.8 mg, 900 μmol), EDCI (138 mg, 720 μmol), and DMAP (88 mg, 720 μmol) were added to 10 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction system was poured into brine, the organic phase was separated, and the aqueous phase was extracted three times with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 2 (305 mg, yellow oil) in a yield of 50%. LC-MS (ESI+): 1019.6 [M+H]+

[0173] 1 H NMR (400MHz, CDCl3) δ6.69(d,J=1.4Hz,2H),6.41(t,J=1.5Hz,1H),5.28(t,J=6.1Hz,1 H),5.21(s,2H),4.72(p,J=7.4Hz,1H),4.12(q,J=7.5Hz,6H),3.49(s,2H),2.75(s,6H ),2.44(t,J=5.8Hz,2H),2.27–2.06(m,5H),2.01–1.82(m,5H),1.77(dq,J=7.2,5.4Hz ,2H),1.72–1.42(m,17H),1.42–1.17(m,32H),1.16–1.02(m,3H),1.00–0.75(m,21H).

[0174] Example 3 Synthesis of Compound 3

[0175] Compound A10 (560 mg, 600 μmol), compound C1 (141.5 mg, 900 μmol), EDCI (138 mg, 720 μmol), and DMAP (88 mg, 720 μmol) were added to 10 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction system was poured into brine, the organic phase was separated, and the aqueous phase was extracted three times with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 3 (270 mg, yellow oil) in a yield of 42%. LC-MS (ESI+): 1073.7 [M+H]+

[0176] 1H NMR (400MHz, CDCl3) δ6.68(d,J=3.0Hz,2H),6.37(t,J=2.9Hz,1H),5.27(m,1H),5.20(s,2H),4.60(p,J=14.4Hz,1H),4.22–4.02(m,6H),3.29(s ,2H),2.61–2.29(m,6H),2.30–2.03(m,6H),2.03–1.80(m,5H),1.79–1. 44(m,22H),1.43–1.19(m,34H),1.19–0.98(m,4H),0.95–0.81(m,24H).

[0177] Example 4 Synthesis of Compound 4

[0178] Compound A10 (561 mg, 600 μmol), compound C2 (143 mg, 900 μmol), EDCI (139 mg, 720 μmol), and DMAP (89 mg, 720 μmol) were added to 10 mL of dichloromethane and stirred at room temperature for 15 hours. The reaction system was poured into brine, the organic phase was separated, and the aqueous phase was extracted three times with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 4 (238 mg, yellow oil) in a yield of 37%. LC-MS (ESI+): 1074.7 [M+H]+

[0179] 1 H NMR (400MHz, CDCl3) δ6.62 (s, 2H), 6.34 (t, J = 2.9 Hz, 1H), 5.21 (tt, J = 12.5 ,2.0Hz,1H),5.20(s,2H),4.71(p,J=14.8Hz,1H),4.20–4.02(m,6H),3.28( s,2H),2.44(td,J=11.7,1.2Hz,2H),2.32(s,8H),2.28–2.05(m,8H),2.04 –1.42(m,24H),1.42–1.19(m,30H),1.19–0.99(m,3H),0.97–0.81(m,21H).

[0180] Example 5 Synthesis of Compound 5

[0181] Step 1: Synthesis of Compound B2

[0182] Compound B1 (5.00 g, 21.7 mmol) was dissolved in 50 mL of tetrahydrofuran, and sodium hydride (960 mg, 23.8 mmol) was added under ice bath conditions. After stirring for 30 minutes, benzyl bromide (2.84 mL, 23.8 mmol) was slowly added, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, 150 mL of ice water was slowly added to the reaction system to quench the reaction, and 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 silica gel column chromatography to obtain compound B2 (6.15 g, yellow oil) in a yield of 88%.

[0183] Step 2: Synthesis of Compound B3

[0184] Compound B2 (6.00 g, 18.7 mmol) was dissolved in 30 mL of methanol and placed in an ice bath. Hydrochloric acid / methanol solution (4.0 M, 70 mL) was slowly added to the reaction system and stirred at room temperature for 6 hours. After the reaction was completed, the mixture was directly dried to obtain compound B3 (5.12 g, light yellow solid) with a yield of 93%.

[0185] Step 3: Synthesis of Compound B4

[0186] Compound B3 (1.50 g, 5.11 mmol) was added to a reaction flask, followed by acetonitrile (25 mL), potassium carbonate (5.89 g, 42.627 mmol) and tert-butyl 4-bromobutyrate (1.37 g, 6.13 mmol), and the mixture was stirred at room temperature for 18 hours after dissolving. The mixture was then filtered, the filter cake was washed with ethyl acetate, 100 mL of water was added to the filtrate, the liquid was separated, the organic phase was separated, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, the organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to give compound B4 (1.80 g, yellow oil) in a yield of 97%. LCMS (ESI+): 363.4 [M+H]+.

[0187] Step 4: Synthesis of Compound B5

[0188] Compound B4 (1.80 g, 4.96 mmol) was dissolved in 10 mL of dichloromethane. A 4.0 M hydrochloric acid / 1,4-dioxane solution (20 mL) was slowly added to the reaction system in an ice bath. The mixture was allowed to warm to room temperature and stirred for 2 hours. After the reaction, the solvent was removed by rotary evaporation to obtain compound B5 (1.80 g, yellow oil) in a 95% yield.

[0189] Step 5: Synthesis of compound 5

[0190] Compound A10 (2.00 g, 2.14 mmol), compound B5 (810 mg, 2.14 mmol), DIEA (420 mg, 3.21 mmol), and DMAP (30 mg, 0.214 mmol) were mixed and dissolved in 20 mL of dichloromethane. DCC (1.33 g, 6.43 mmol) was added under ice-cooling. The reaction system was allowed to warm to room temperature and stirred for 18 hours. After the reaction, 20 mL of water was added, and the organic phase was separated. The aqueous phase was extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain compound 5 (1.90 g, yellow oil) in a yield of 72%.

[0191] 1 H NMR (400MHz, CDCl3): δ7.34-7.27(m,5H),6.46(s,2H),6.38(s,1H),5.37(m,1H),5.02(s,2H),4.64-4.62(m,1H),4.54(s,2H),4.09(t,J= 6.4Hz,4H),3.98(t,J=6.0Hz,2H),3.92(t,J=6.4Hz,2H),3.59(t,J=6.0Hz,2H),3.50-3.48(m,2H),2.62(t,J=6.0Hz,2H),2.49(d,J=6.4Hz ,2H),2.40-2.31(m,6H),2.13-2.05(m,2H),1.97-1.90(m,4H),1.84-1.78(m,6H),1.72-1.67(m,5H),1.61-1.58(m,5H),1.50-1.42(m,8H ),1.37-1.34(m,4H),1.25(m,20H),1.18-1.08(m,10H),1.02(s,3H),0.96(m,1H),0.92(d,J=6.4Hz,3H),0.89-0.86(m,12H),0.68(s,3H).

[0192] Example 6 Synthesis of Compound 6

[0193] Potassium carbonate (0.47 mmol) and 5 mL of dichloromethane were added to a reaction flask, followed by chlorosulfonyl isocyanate (CSI) (0.32 mmol). The mixture was stirred thoroughly, and 250 mg of compound 5 (0.21 mmol) was added. The mixture was heated to 30°C for 20 hours, then cooled to 0°C, and 4 mL of methanol and sodium hydroxide (1 mmol) were added. The mixture was stirred at room temperature for 1 hour. The solvent was removed by rotary evaporation, and 10 mL of water was added. After neutralization with 1N hydrochloric acid, 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 silica gel column chromatography to obtain compound 6 (166 mg, yellow oil) in a 70% yield. LC-MS (ESI+): 1131.8 [M+H]+.

[0194] 1 H NMR (400MHz, CDCl3): δ6.45(s,2H),6.33(s,1H),5.39(m,1H),4.64-4.62(m,1H),4.54(s,2H),4.09(t,J=6. 2Hz,4H),3.98(m,2H),3.92(m,2H),3.59(t,J=6.4Hz,2H),3.50-3.48(m,2H),2.61(t,J=5.8Hz,2H),2.48(d ,J=6.0Hz,2H),2.40-2.31(m,6H),2.13-2.05(m,2H),1.97-1.90(m,4H),1.84-1.78(m,6H),1.72-1.40(m,1 8H),1.39-1.05(m,34H),1.02(s,3H),0.96(m,1H),0.92(d,J=6.4Hz,3H),0.94-0.84(m,12H),0.68(s,3H).

[0195] Example 7: Preparation and Characterization of mRNA-LNP

[0196] 1. Materials and Instruments

[0197] Table 1 Main experimental consumables

[0198] Table 2 Main experimental equipment

[0199] Table 3 Other main reagents

[0200] 2. Experimental Plan

[0201] Preparation of mRNA-LNPs

[0202] The lipid components were mixed in an ethanol solution at a molar ratio of 65.7:32.8:1.5. Firefly luciferase mRNA (Luc-mRNA) was dissolved in 25 mM sodium acetate buffer, pH 4.0, to a final concentration of 135 ng / uL. The aqueous and ethanolic phases were mixed using a microfluidic device at a mixing flow rate of 9 mL / min for the aqueous phase and 3 mL / min for the ethanolic phase. The prepared mRNA-LNPs were dialyzed and concentrated by ultrafiltration into a 20 mM Tris / 25 mM sodium acetate / 87% sucrose buffer, pH 7.5. The experimental samples were sterile filtered.

[0203] Characterization of mRNA-LNPs

[0204] 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.

[0205] Table 4 Physical and chemical parameters of Luc-mRNA-LNP

[0206] *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.

[0207] 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, an average particle size in the range of about 60-102 nm, a PDI of less than 0.15, a good polydispersity coefficient, and a zeta potential between -5mV–5mV. The encapsulation efficiency of LNP for mRNA is greater than 90%, which is significantly better than that of the control group LNP.

[0208] Example 8 mRNA-LNP in vitro cell transfection activity

[0209] The expression level of firefly luciferase Luc was detected by fluorescence microscopy to evaluate the transfection activity of mRNA-LNP on HEK293T cells. 5 HEK293T cell solution of 100 cells / mL was inoculated into 96-well cell culture plates at a volume of 200 μL / well. After 24 hours, 100 ng of Luc-mRNA-LNP was transfected into each well and the cell culture plates were placed in a 37°C, 5% CO2 cell culture incubator. The negative control group was transfected with an equal volume of PBS. After 24 hours, the cells were cultured according to the Fire-Lumi TM Bioluminescence detection was performed according to the instructions of the luciferase detection kit, 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 Luc-mRNA in cells, and the expression level is significantly better than that of the control group.

[0210] Example 9 mRNA-LNP Animal Immunization Test

[0211] 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-PEG2000 = 65.7:32.8:1.5 molar ratio for mRNA encapsulation. The specific formulation is shown in Table 4. Female BALB / c mice aged 6-8 weeks were randomly divided into groups of 5 and immunized via intramuscular injection of the hind legs. Immunizations were performed on days 0 and 14, with a dose of 5 μg mRNA-LNP per mouse. On day 28, blood was collected and serum was separated. Specific antibody titers against the SARS-CoV-2 S protein antigen were measured by ELISA (enzyme-linked immunosorbent assay), and specific CD8+ and CD4+ T cell immune responses in the spleen were measured by ICS (intracellular cytokine staining). The results of antibody titer, CD8+T cell immunity and CD4+T cell immune response are shown in Figures 2, 3 and 4 respectively. After the three-component LNP constructed by the lipid compound provided by the present invention delivered the new crown mRNA vaccine, the antibody titer, CD8+T cell and CD4+T cell immune response induced in mice were significantly better than the three-component LNP composed of the control group compound. The results show that the immunogenicity of the mRNA vaccine composition formed by the lipid nanoparticles provided by the present invention is higher than that of the control group.

[0212] 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. The lipid compound represented by formula (I): or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof; wherein, Ar is an aryl or heteroaryl group; the aryl or heteroaryl group is optionally substituted with a group selected from the following: C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl, halogen, alkoxy, alkylthio, NR4R4’, R4-C(O)-, R4-C(O)O-; G1, G2 and G3 are each independently selected from a bond, an optionally substituted C1-C 20 alkylene, an optionally substituted C2-C 20 alkenylene, an optionally substituted C2-C 20 alkynylene, an optionally substituted carbocyclic moiety, an optionally substituted arylene; L1, L2, L3, L4, L5, and L6 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 )-, -S-S-, -OC(=O)S-, -SC(=O)O-, -N(R a )C(=O)N(R b )-; 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 alkynyl, one or more of -CH2- therein may optionally be replaced by -O-, -S-, -NR a -, carbocyclic group, aryl, heteroaryl, and / or heterocyclic group; Meanwhile, another one of R1, R2, and R3 is selected from a steroid group; Meanwhile, the third 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 -guanidyl; wherein, the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidyl 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, R c O-C1-C 20 alkyl; R4 and R4’ are each independently selected from H, C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic group and / or heterocyclic alkyl; R4” is selected from C1-C 20 alkylene, C2-C 20 alkenylene, C2-C 20 alkynylene; R a 、R b and R c 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 carbocyclic group, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclic group, optionally substituted heterocyclic alkyl; m, n, and p are each independently selected from 1, 2, or 3; q is selected from 0 or 1.

2. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to claim 1; wherein Ar is an aryl group; the aryl group is optionally substituted with a group selected from the following: C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl group, halogen, alkoxy group, alkylthio group, NR4R4’, R4-C(O)-, R4-C(O)O-; G1, G2 and G3 are each independently selected from a bond, an optionally substituted C1-C 20 alkylene, an optionally substituted C2-C 20 alkenylene, an optionally substituted C2-C 20 alkynylene, an optionally substituted carbocyclic moiety, an optionally substituted arylene; L1, L2, L3, L4, L5, and L6 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 )-, -S-S-, -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, The C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 one or more -CH2- in alkynyl may optionally be replaced by O, S, -NR a -, carbocyclic group, aryl, heteroaryl, and / or heterocyclic group; R2 is selected from a steroid group; R3 is selected from -(R4”) q -NR a R b 、-(R4”) q -nitrogen-containing heteroaryl, -(R4”) q -nitrogen-containing heterocyclic group, -(R4”) q -guanidyl; wherein, the nitrogen-containing heteroaryl, nitrogen-containing heterocyclic group and guanidyl 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, R c O-C1-C 20 alkyl; R4 and R4’ are each independently selected from H, C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic group and / or heterocyclic alkyl; R4” is selected from C1-C 20 alkylene, C2-C 20 alkenylene, C2-C 20 alkynylene; R a 、R b and R c 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 carbocyclic group, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclic group, optionally substituted heterocyclic alkyl; m, n, and p are each independently selected from 1, 2, or 3; q is selected from 0 or 1.

3. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to claim 2; wherein Ar is phenyl; the phenyl is optionally substituted with a group selected from the following: C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl, halogen, alkoxy, alkylthio, NR4R4’, R4-C(O)-, R4-C(O)O-; G1, G2, and G3 are each independently selected from a bond, an optionally substituted C1-C 20 alkylene group, an optionally substituted C2-C 20 alkenylene group, an optionally substituted C2-C 20 alkynylene group each time they appear; L1, L2, L3, L4, L5, and L6 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 )-, -S-S-, -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, The C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, one or more of the -CH2- may optionally be replaced by O, S, -NR a -, carbocyclic group, aryl, heteroaryl, and / or heterocyclic group; R2 is selected from a steroid group; R3 is selected from -(R4”) q -NR a R b 、-(R4”) q -a nitrogen-containing heteroaryl, -(R4”) q -a nitrogen-containing heterocyclic group; wherein, the nitrogen-containing heteroaryl and nitrogen-containing heterocyclic group are optionally substituted by one or more groups selected from: 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, R c O-C1-C 20 alkyl; R4 and R4’ are each independently selected from H, C1-C 20 alkyl, C2-C 20 alkenyl, C2-C 20 alkynyl, carbocyclic group, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic group and / or heterocyclic alkyl; R4” is selected from C1-C 20 alkylene, C2-C 20 alkenylene, C2-C 20 alkynylene; R a 、R b and R c 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 carbocyclic group, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heterocyclic group, optionally substituted heterocyclic alkyl; 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 any one of the preceding claims, selected from or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof; wherein L1, L2, L3, L4, L5, L6, G1, G2, G3, R1, R2, R3, m, n, and p are as defined in any one of claims 1-3.

5. The compound according to any one of the preceding claims, selected from or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof; Among them, L1, L2, L3, L4, L5, L6, G1, G2, G3, R1, R2, R3, m, n, and p are as defined in any one of claims 1-3.

6. The compound according to any one of the preceding claims, selected from or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof; Among them, R1, R2, and R3 are as defined in any one of claims 1-3, and r, s, t, and u are each independently selected from integers from 1 to 10.

7. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to any one of the preceding claims; Among them, The steroid in the steroid group is selected from naturally occurring steroids or their analogs; preferably, including phytosterols and zoosterols, or their analogs; more preferably, selected from cholesterol and its derivatives.

8. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to any one of the preceding claims; Among them, The steroid 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, hydroxy, 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 compound group is selected from: wherein R' is C 1-20 alkyl.

9. The compound according to any one of the preceding claims, selected from: or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof, wherein R1, R3 are as defined in any one of claims 1-3, and r, s, t, and u are each independently selected from integers from 1 to 10.

10. The compound according to any one of the preceding claims, selected from: or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof, wherein R1, R3 are as defined in any one of claims 1-3, and r, s, t, and u are each independently selected from integers from 1 to 10.

11. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to any one of the preceding claims; wherein R1 is selected from optionally substituted C1-C 20 alkyl; wherein, One or more -CH2- in the C1-C 20 alkyl group may optionally be replaced by O, S, -NR a -, or a carbocyclic group.

12. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to any one of the preceding claims; wherein R1 is selected from 13. The compound or a stereoisomer, tautomer, and pharmaceutically acceptable salt thereof according to any one of the preceding claims; wherein R3 is selected from -(R4”) q -NR a R b 、-(R4”) q -a 5- or 6-membered nitrogen-containing heteroaryl, -(R4”) q -a 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, hydroxy, mercapto, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy, R c O-C1-C6 alkyl; R4” is selected from C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene; R a and R b each independently selected from H and C1-C6 alkyl; R c selected from H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, phenyl-C1-C6 alkyl; q is selected from 0 or 1.

14. A compound or its stereoisomer, tautomer, and pharmaceutically acceptable salt according to any one of the preceding claims; wherein R3 is selected from -R4”-NR a R b , -R4”-a 5- or 6-membered nitrogen-containing heterocyclic group; wherein, The 5- or 6-membered nitrogen-containing heterocyclic group is optionally substituted with a group selected from the following: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogen, hydroxy, mercapto, cyano, nitro, amino, C1-C6 acyl, C1-C6 acyloxy, R c O-C1-C6 alkyl; R4” is selected from C1-C6 alkylene; R a and R b each independently selected from H, C1-C6 alkyl; R c selected from H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, benzyl.

15. The compound according to claim 1, selected from: or its stereoisomer, tautomer, and pharmaceutically acceptable salt thereof.

16. A lipid nanoparticle comprising the lipid compound or its stereoisomer, tautomer, and pharmaceutically acceptable salt according to any one of claims 1-15.

17. The lipid nanoparticle according to claim 16, further comprising a phospholipid and / or a polyethylene glycol lipid.

18. The lipid nanoparticle according to claim 17, further comprising a therapeutic agent and / or a prophylactic agent.

19. The lipid nanoparticle according to claim 18, wherein the therapeutic agent and / or prophylactic 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 the compound according to any one of claims 1-15 and / or the lipid nanoparticle according to claims 16-19 in the preparation of a delivery system for a therapeutic agent and / or a prophylactic agent.

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