Lipid compound and lipid nanoparticle for delivery

Abstract

The present invention relates to a lipid compound and lipid nanoparticle for delivery. Disclosed are a compound having a structural formula as shown in formula (I), and a pharmaceutically acceptable salt thereof or a stereoisomer thereof. Further disclosed is a nanoparticle composition comprising the compound, or the pharmaceutically acceptable salt thereof or the stereoisomer thereof. The nanoparticle of the present application can efficiently deliver a drug or vaccine to the lungs and spleen of a mammal, thereby achieving the therapeutic or preventive purpose of the drug or vaccine.

Description

Lipid compounds and lipid nanoparticles for delivery Technical Field

[0001] This application relates to the field of biotechnology, specifically to lipid compounds and lipid nanoparticle compositions for the delivery of active ingredients such as drugs and vaccines. Background Technology

[0002] mRNA vaccines have achieved remarkable success in combating the COVID-19 pandemic due to their high efficiency in design, research and development, and production. Theoretically, mRNA vaccines can achieve good therapeutic effects in infectious diseases, protein replacement, and tumors. However, efficient mRNA delivery remains the biggest bottleneck in clinical translation.

[0003] Lipid-containing nanoparticles, or lipid nanoparticles, liposomes, and lipid complexes, have been proven to be effective transport carriers for bioactive substances such as small molecule drugs, proteins, and nucleic acids that enter and / or enter cells. Lipid nanoparticles (LNPs) are small vesicles formed from one or more lipid components that can effectively compress and deliver various nucleic acid molecules, from DNA and RNA to chromosomes and even cells. LNPs are conducive to large-scale production due to their well-defined construction schemes and ease of modification with targeted ligands.

[0004] However, current liver-specific intravascular coagulation (LNPs) are primarily used for liver delivery, and their effectiveness is less than satisfactory when addressing delivery needs to other organs such as the lungs and spleen. Given the diversity of indications for lung and spleen diseases, there is a significant unmet clinical need; therefore, developing LNPs with targeted delivery capabilities for the lungs and spleen is of paramount importance. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a lipid compound and lipid nanoparticles containing the compound, wherein the LNP can achieve targeted delivery to the lungs and spleen.

[0006] To solve the above problems, the technical solution adopted by the present invention is as follows:

[0007] 1. A compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0008]

[0009] Formula (I)

[0010] Wherein, R1 is independently selected from C1-C14 alkyl groups;

[0011] R2 is independently selected from hydrogen or C1-C14 alkyl groups;

[0012] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, and X2 and Y2 are not simultaneously C=O or O;

[0013] m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0014] n is independently selected from 1, 2, 3, 4, or 5.

[0015] 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein n is independently selected from 1, 2, 3, 4, preferably 2 or 3.

[0016] 3. The compound according to any one of items 1-2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein m is independently selected from 2, 3, 4, 5, 6, 7 or 8, preferably 3, 4, 5, 6 or 7.

[0017] 4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R1 is independently selected from a straight-chain alkyl group of C6-C12, preferably a straight-chain alkyl group of C7-C9;

[0018] R2 is independently selected from hydrogen or a C6-C12 straight-chain alkyl group, preferably hydrogen or a C7-C9 straight-chain alkyl group.

[0019] 5. The compound according to any one of items 1 to 4, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R1 is independently selected from a straight-chain alkyl group of C6-C12, preferably a straight-chain alkyl group of C7-C9;

[0020] R2 is independently selected from C6-C12 straight-chain alkyl groups, preferably C7-C9 straight-chain alkyl groups;

[0021] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, X2 and Y2 are not simultaneously C=O or O. Preferably, X1 and X2 are both C=O, and Y1 and Y2 are both O.

[0022] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0023] n is independently selected from 2 or 3.

[0024] 6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the compound of (I) is selected from...

[0025] Compound 1:

[0026] ,

[0027] Compound 2: ,

[0028] Compound 3: ,

[0029] Compound 4:

[0030] ,

[0031] Compound 5:

[0032] ,

[0033] Compound 6: or

[0034] Compound 7:

[0035] .

[0036] 7. A lipid nanoparticle composition comprising a lipid component, said lipid component comprising any one of claims 1-6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0037] 8. The lipid nanoparticle composition according to claim 7, wherein the lipid component further comprises phospholipids.

[0038] 9. The lipid nanoparticle composition according to claim 8, wherein the phospholipid is selected from one or more of the following compounds:

[0039] Dilauroyl lecithin (DLPC)

[0040] Dimyristic phosphatidylcholine (DMPC)

[0041] Dioleoyl lecithin (DOPC)

[0042] Dipalmitoylphosphatidylcholine (DPPC)

[0043] Distearate phosphatidylcholine (DSPC)

[0044] Dioleoylphosphatidylcholine (DUPC)

[0045] Palmitoyl oleoyl phosphatidylcholine (POPC)

[0046] 1,2-Di-O-octadecyl-sn-glycerol-3-phosphate choline (18:0 Diether PC)

[0047] 1-Oleoyl-2-cholesterol dimethylsuccinate-sn-glycerol-3-phosphate choline (OChemsPC)

[0048] l-Hexadecyl-sn-glycerol-3-phosphate choline (C16 Lyso PC)

[0049] 1,2-Divinyl-sn-glycerol-3-phosphate choline,

[0050] 1,2-Diarylac-sn-glycerol-3-phosphocholine,

[0051] 1,2-Dioleoyl-SN-glycerol-3-phosphoethanolamine (DOPE)

[0052] 1,2-Distearate-sn-glycerol-3-phosphate ethanolamine,

[0053] 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine,

[0054] 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine,

[0055] 1,2-Diaryl-sn-glycerol-3-phosphate ethanolamine

[0056] 1,2-Dithiohexaenoic acid-sn-glycerol-3-phosphate ethanolamine,

[0057] 1,2-Dioleoyl-sn-glycerol-3-phosphate-(1-glycerol)sodium salt (DOPG) or sphingomyelin,

[0058] 10. The nanoparticle composition according to item 8, wherein the phospholipid is DOPE.

[0059] 11. The nanoparticle composition according to item 8, wherein the phospholipid is DSPC.

[0060] 12. The nanoparticle composition according to any one of items 7-11, wherein the lipid component further comprises structural lipids.

[0061] 13. The nanoparticle composition according to claim 12, wherein the structural lipid is selected from one or more of cholesterol, coccosterol, sitosterol, ergosterol, and stigmasterol.

[0062] 14. The nanoparticle composition according to item 12, wherein the structural lipid is cholesterol.

[0063] 15. The nanoparticle composition according to any one of claims 7-14, wherein the lipid component further comprises PEG lipids.

[0064] 16. The nanoparticle composition according to claim 15, wherein the PEG lipid is selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol or PEG-modified dialkylglycerol.

[0065] 17. The nanoparticle composition according to any one of claims 7-16, wherein the lipid component further comprises cationic and / or ionizable lipids.

[0066] 18. The nanoparticle composition according to any one of claims 7-17, further comprising a therapeutic agent and / or a preventive agent selected from vaccines or compounds capable of inducing an immune response, nucleic acids,

[0067] Preferably, the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

[0068] 19. The nanoparticle composition according to any one of items 7-18, wherein the encapsulation efficiency of the therapeutic agent and / or preventive agent is ≥50%; or ≥80%; or ≥90%; or 100%.

[0069] 20. The nanoparticle composition according to any one of items 7-19, wherein the average particle size of the nanoparticle composition is 50 nm to 110 nm.

[0070] 21. The nanoparticle composition according to any one of items 7-20, wherein the dispersibility index of the nanoparticle composition is 0.003-0.30.

[0071] 22. Use of the compound of any one of items 1-6 in the preparation of lipid nanoparticle compositions.

[0072] 23. A pharmaceutical composition comprising the nanoparticle composition described in any one of claims 7-21 and a pharmaceutically acceptable carrier.

[0073] 24. A method of delivering a therapeutic and / or preventive agent to mammalian cells, the method comprising administering to a subject the nanoparticle composition of any one of items 7-21 or the pharmaceutical composition of item 23, the administration comprising contacting cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and / or preventive agent to the cells.

[0074] 25. The method according to item 24, wherein the mammalian cell is in a mammal.

[0075] 26. The method according to item 24 or 25, wherein the mammal is a human.

[0076] 27. The method according to any one of items 24-26, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0077] 28. A method for producing a target polypeptide in mammalian cells, the method comprising contacting the cells with a nanoparticle composition of any one of items 7-21 or a pharmaceutical composition of item 23 to deliver a therapeutic and / or preventive agent to the cells, wherein the therapeutic and / or preventive agent is mRNA encoding the target polypeptide, thereby enabling the mRNA to be translated in the cells to produce the target polypeptide.

[0078] 29. The method according to item 28, wherein the mammalian cell is in a mammal.

[0079] 30. The method described in item 27 or 28, wherein the mammal is a human.

[0080] 31. The method according to any one of items 28-30, wherein the nanoparticle composition or pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0081] 32. A method for specifically delivering a therapeutic and / or preventive agent to a mammalian organ, the method comprising administering to a mammal any of the nanoparticle compositions described in any one of items 7-21 or the pharmaceutical composition described in item 23, the administration comprising contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or preventive agent to the organ, preferably the lung and spleen.

[0082] 33. The method according to item 32, wherein the mammal is a human.

[0083] 34. The method according to item 32 or 33, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0084] 35. The method according to any one of items 32-34, wherein the mammal is pretreated 24 hours or less prior to the contact or administration step.

[0085] 36. The method according to any one of claims 32-35, wherein the mammal is pretreated for about one hour prior to the contact or administration step.

[0086] 37. A method for treating a disease or ailment in a mammal, the method comprising administering to the mammal a therapeutically effective amount of any one of items 7-21 of the nanoparticle composition or the pharmaceutical composition of item 23.

[0087] 38. The method according to item 37, wherein the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity.

[0088] 39. The method according to item 37 or 38, wherein the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, or metabolic diseases.

[0089] 40. The method according to any one of items 37-39, wherein the mammal is a human.

[0090] 41. The method according to any one of items 37-40, wherein the nanoparticle composition or pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0091] The beneficial effects of adopting the above technical solution are as follows:

[0092] The compounds of this application can be used to prepare lipid nanoparticles. Nanoparticle compositions containing the compounds provided in this application can encapsulate and deliver therapeutic / preventive agents, safely delivering the therapeutic / preventive agents to the lungs and spleen to achieve high expression and exert the therapeutic / preventive effects.

[0093] The lipid nanoparticles prepared in this application have small average particle size, high encapsulation efficiency, and high expression, and have broad application prospects in the field of drug delivery. Attached Figure Description

[0094] Figure 1 shows the Luciferase fluorescence intensity of LNPs of compounds 1, 3, and 4 6 hours after intravenous injection.

[0095] Figure 2 shows the anatomical diagrams of the lungs and stomach of mice that died after intravenous injection of LNP of compound 2. Detailed Implementation

[0096] To make the objectives, technical solutions, and advantages of the present invention clearer, the invention will be described clearly and completely below in conjunction with specific embodiments.

[0097] Terms and Definitions

[0098] As used herein, the term "alkyl" refers to a hydrocarbon comprising one or more carbon atoms (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, or more carbon atoms), which may optionally be substituted. The term "C1-C14 alkyl" refers to a saturated hydrocarbon of 1-14 carbon atoms, either straight-chain or branched. Unless otherwise stated, alkyl as described herein refers to both unsubstituted and substituted alkyl groups.

[0099] Unless otherwise specified, alkyl groups may optionally be substituted. Optional substituents may be selected from, but are not limited to, halogen atoms (e.g., chlorine, bromine, fluorine, or iodine), carboxylic acids (e.g., -C(O)OH), alcohols (e.g., hydroxyl, -OH), esters (e.g., -C(O)OR or -OC(O)R), aldehydes (e.g., -C(O)H), carbonyl groups (e.g., -C(O)R, or represented by C=O), acyl halides (e.g., -C(O)R, where R is a halide selected from bromides, fluorides, chlorides, and iodides), carbonates (e.g., -OC(O)OR), alkoxy groups (e.g., -OR), acetals, phosphates, thiols (e.g., -SH), sulfoxides (e.g., -S(O)R), sulfurous acids (e.g., -S(O)OH), sulfonic acids (e.g., -S(O)2OH), thiols (e.g., -C(S)H), sulfates, sulfonyl groups (e.g., -S(O)2), and thiols (e.g., -C(S)H). - ), amides (e.g., -C(O)NR2 or -N(R)C(O)R), azides (e.g., -N3), nitro groups (e.g., -NO2), cyano groups (e.g., -CN), isocyano groups (e.g., -NC), acyloxy groups (e.g., -OC(O)R), amino groups (e.g., -NR2, -NRH, or -NH2), carbamoyl groups (e.g., -OC(O)NR2, -OC(O)NRH, or -OC(O)NH2), sulfonamides, alkyl groups, alkenyl groups, and cyclic groups (e.g., carbocyclic or heterocyclic groups). In any of the foregoing, R is an alkyl or alkenyl group as defined herein. In some embodiments, the substituent itself may be further substituted with one, two, three, four, five, or six substituents as defined herein. For example, a C6-C12 alkyl group may be further substituted with 6, 7, 8, 9, 10, 11, or 12 substituents as described herein.

[0100] As used herein, the term "contact" refers to establishing a physical connection between two or more entities. For example, contacting a mammalian cell with a nanoparticle composition means that the mammalian cell and the nanoparticles share a physical connection. Methods for contacting cells with external entities in vivo and in vitro are well known in the field of biology. For example, nanoparticle compositions can be contacted with mammalian cells placed within mammals via various routes of administration (e.g., intravenous, intramuscular, intradermal, and subcutaneous), and can involve various amounts of the nanoparticle composition. Furthermore, the nanoparticle composition can contact more than one mammalian cell.

[0101] As used herein, “encapsulation ratio” refers to the amount of therapeutic and / or preventive agent that is part of the nanoparticle composition, relative to the total amount of therapeutic or preventive agent used in the preparation of the nanoparticle composition. For example, if 97 mg of therapeutic and / or preventive agent is encapsulated in the nanoparticle composition out of a total of 100 mg of therapeutic and / or preventive agent initially provided to the composition, the encapsulation ratio can be 97%. As used herein, “encapsulation” can mean complete, substantial, or partial encapsulation, closure, enclosure, or sealing.

[0102] As used in this article, “expression” of a nucleic acid sequence refers to the translation of mRNA into a polypeptide or protein and / or post-translational modifications of the polypeptide or protein.

[0103] As used herein, the term "in vitro" refers to events that occur in an artificial environment, such as in a test tube or reaction vessel, in a cell culture, in a petri dish, etc., rather than events that occur within a living organism (e.g., an animal, plant, or microorganism).

[0104] As used in this article, the term "in vivo" refers to events that occur within an organism, such as an animal, plant, or microorganism or its cells or tissues.

[0105] As used herein, the term "ex vivo" refers to an event that occurs outside an organism (e.g., an animal, plant, or microorganism, or its cells or tissues). Ex vivo events can occur in environments with minimal alteration from the natural (e.g., internal) environment.

[0106] As used herein, the term "isomer" refers to any geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer, or diastereomer of a compound. A compound may contain one or more chiral centers and / or double bonds, and thus may exist in stereoisomeric form, such as double-bonded isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis-trans isomers). This application covers any and all isomers of the compounds described herein. Mixtures of enantiomers and stereoisomers of compounds, and methods for resolving them into their component enantiomers or stereoisomers, are well known.

[0107] As used herein, a "lipid component" is a component of a nanoparticle composition comprising one or more lipids. For example, a lipid component may include one or more cationic / ionizable lipids, PEGylated lipids, structured lipids, or other lipids such as phospholipids.

[0108] As used herein, “modified” means non-natural. For example, RNA can be modified RNA. That is, RNA may include one or more non-naturally occurring nucleobases, nucleosides, nucleotides, or linkers. A “modified” substance may also be referred to herein as a “modified” substance. Such a substance may be chemically, structurally, or functionally modified or altered. For example, the types of modified nucleobases may include one or more non-naturally occurring substitutions.

[0109] As used herein, “method of administration” may include intravenous, intramuscular, intradermal, subcutaneous, or other methods of delivering the composition to a subject. Any method of administration may be chosen for targeted delivery (e.g., specific delivery) to a specific area or system of the body.

[0110] As used herein, a “nanoparticle composition” is a composition comprising one or more lipids. The particle size of a nanoparticle composition is typically on the order of micrometers or smaller and may include a lipid bilayer. Nanoparticle compositions include lipid nanoparticles (LNPs), liposomes (e.g., lipid vesicles), and lipid complexes. For example, a nanoparticle composition may be a liposome having a lipid bilayer with a diameter of 500 nm or smaller.

[0111] As used herein, the term "enhanced delivery" refers to the delivery of a greater (e.g., at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times) of therapeutic and / or preventative drugs to a target tissue (e.g., mammalian lungs) via nanoparticles. This can be achieved by comparing the amount of protein produced in the tissue to the weight of the tissue, the amount of therapeutic and / or preventative agents in the tissue to the weight of the tissue, the amount of protein produced within the tissue to the total amount of protein in the tissue, or the amount of therapeutic and / or preventative agents in the tissue to the total amount of therapeutic and / or preventative agents in the tissue. It should be understood that enhanced delivery of nanoparticles to target tissues does not need to be determined in treated subjects but can be determined in alternatives such as animal models (e.g., rat models). In some embodiments, when the nanoparticle composition comprises a compound of formula (I), it has substantially the same level of enhanced delivery regardless of the route of administration.

[0112] As used herein, the term "specific delivery" or "specific transport" refers to the delivery of more (e.g., at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times) of therapeutic and / or preventative drugs to a target tissue (such as the mammalian lung) compared to non-target tissues. The level of nanoparticle delivery to a specific tissue can be measured by comparing the weight of proteins produced in the tissue to the weight of the tissue, comparing the amount of therapeutic and / or preventative drugs in the tissue to the weight of the tissue, comparing the weight of proteins produced in the tissue to the total weight of proteins in the tissue, or comparing the amount of therapeutic and / or preventative drugs in the tissue to the total amount of therapeutic and / or preventative drugs in the tissue.

[0113] As used in this article, “naturally existing” means existing naturally without artificial intervention.

[0114] As used herein, "PEG lipid" or "PEGylated lipid" refers to lipids containing polyethylene glycol.

[0115] The term “pharmaceutically acceptable” is used herein to refer to those compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues to the extent of reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0116] As used herein, the phrase "pharmaceutically acceptable excipient" means any component other than the compounds described herein (e.g., a medium capable of suspending, complexing, or dissolving an active compound) and is substantially non-toxic and non-inflammatory to the patient. Excipients may include, for example: anti-adhesives, antioxidants, adhesives, coatings, anti-compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavorings, fragrances, flow enhancers (flow improvers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and hydrated water. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (diacid), calcium stearate, crosslinked carboxymethyl cellulose, crosslinked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethyl cellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methyl cellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silica, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E (α-tocopherol), vitamin C, xylitol, and other substances disclosed herein.

[0117] In this application, for convenience, the structural formulas of the compounds represent certain isomers; however, this application includes all isomers, such as geometric isomers, optical isomers based on asymmetric carbons, stereoisomers, tautomers, etc. It should be understood that not all isomers may have the same level of activity. Furthermore, the compounds represented by the structural formulas of the compounds described in this application may exhibit crystal polymorphism. Note that any crystal form, mixture of crystal forms, or its anhydrides or hydrates are included within the scope of this application.

[0118] The nanoparticle compositions of this application may also comprise salts of one or more compounds. The salts may be pharmaceutically acceptable. As used herein, a "pharmaceutically acceptable salt" refers to a derivative of the disclosed compound in which the parent compound is modified by converting an existing acid or base moiety into its salt form (e.g., by reacting the free base with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; bases or organic salts of acidic residues such as carboxylic acids, etc. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disaccharide, dodecyl sulfate, ethanesulfonate, fumarate, glucoheponicate, glucoheponicate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, palmitate, pectate, 3-phenylpropionate, phosphate, picrate, neopentyl ester, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, etc.

[0119] Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Pharmaceutically acceptable salts of this application include, for example, conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of this application can be synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Typically, these salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, in an organic solvent, or in a mixture of both. Non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are typically preferred.

[0120] As used herein, "phospholipid" is a lipid comprising a phosphate ester moiety and one or more carbon chains, such as unsaturated fatty acid chains. Phospholipids may contain one or more (e.g., double or triple) bonds (e.g., one or more unsaturated bonds). Certain phospholipids can facilitate fusion with membranes. For example, cationic phospholipids can interact with one or more negatively charged phospholipids in a membrane (e.g., a cell membrane or intracellular membrane). Fusion of phospholipids with membranes can allow one or more elements of a lipid-containing composition to cross the membrane, thereby allowing, for example, the delivery of one or more elements to the cell.

[0121] As used herein, the term “polypeptide” or “target polypeptide” refers to a polymer of amino acid residues typically linked by peptide bonds, which may be produced naturally (e.g., isolated or purified) or synthetically.

[0122] As used herein, “RNA” refers to ribonucleic acid, which may be naturally occurring or non-naturally occurring. For example, RNA may include modified and / or non-naturally occurring components, such as one or more nucleobases, nucleosides, nucleotides, or linkers. RNA may include cap structures, chain-terminating nucleosides, stem-loops, polyA sequences, and / or polyadenylation signals. RNA may have a nucleotide sequence encoding a target polypeptide. For example, RNA may be messenger RNA (mRNA). Translation of mRNA encoding a specific polypeptide, such as in vivo translation of mRNA within mammalian cells, can produce the encoded polypeptide. RNA may be selected from the unrestricted group, including small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), double-stranded RNA (dsRNA), small hairpin RNA (shRNA), mRNA, and mixtures thereof.

[0123] As used herein, in the context of nanoparticle compositions, “particle size” or “average particle size” refers to the average diameter of the nanoparticle composition.

[0124] As used herein, the terms “subject” or “patient” mean any organism to which the composition according to this application may be administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include animals (e.g. mammals, such as mice, rats, rabbits, non-human primates and humans) and / or plants.

[0125] The terms "therapeutic agent" or "preventive agent" refer to any pharmaceutical agent that, when administered to a subject, has a therapeutic, diagnostic, and / or preventive effect and / or induces the desired biological and / or pharmacological action. Therapeutic agents are also known as "active agents" or "active components." Such substances include, but are not limited to, cytotoxins, radioactive ions, chemotherapeutic agents, small molecule drugs, proteins, and nucleic acids.

[0126] As used herein, the term "treatment" refers to partial or complete relief, mitigation, improvement, alleviation, delay of its onset, inhibition of its progression, reduction of its severity, and / or reduction of the incidence of one or more of its symptoms or features. A specific infection, disease, symptom, and / or condition. For example, "treatment" of cancer can refer to inhibiting tumor survival, growth, and / or spread. To reduce risk, treatment may be administered to subjects who do not exhibit disease, symptom, and / or condition and / or to subjects who only exhibit early signs of disease, symptom, and / or condition. A pathological development associated with a disease, symptom, and / or condition.

[0127] This application discloses compounds of formula (I), or pharmaceutically acceptable salts thereof, or stereoisomers thereof.

[0128]

[0129] Formula (I)

[0130] Wherein, R1 is independently selected from C1-C14 alkyl groups;

[0131] R2 is independently selected from hydrogen or C1-C14 alkyl groups;

[0132] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, and X2 and Y2 are not simultaneously C=O or O;

[0133] m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0134] n is independently selected from 1, 2, 3, 4, or 5.

[0135] In some preferred embodiments, R1 is independently selected from C6-C12 alkyl groups, such as C6-C11 alkyl groups, C6-C10 alkyl groups, C6-C9 alkyl groups, C6-C8 alkyl groups, C6-C7 alkyl groups, and C7-C9 alkyl groups. For example, R1 is C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, and C12 alkyl.

[0136] In some preferred embodiments, R1 is independently selected from C6-C12 straight-chain alkyl groups, such as those selected from C6-C11, C6-C10, C6-C9, C6-C8, C6-C7, and C7-C9. For example, R1 is a C6 straight-chain alkyl group, a C7 straight-chain alkyl group, a C8 straight-chain alkyl group, a C9 straight-chain alkyl group, a C10 straight-chain alkyl group, a C11 straight-chain alkyl group, or a C12 straight-chain alkyl group.

[0137] In some preferred embodiments, R2 is independently selected from H or C6-C12 alkyl groups, such as H, C6-C11 alkyl groups, C6-C10 alkyl groups, C6-C9 alkyl groups, C6-C8 alkyl groups, C6-C7 alkyl groups, and C7-C9 alkyl groups. For example, R2 is H, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, or C12 alkyl.

[0138] In some preferred embodiments, R2 is independently selected from H or C6-C12 straight-chain alkyl groups, such as H, C6-C11 straight-chain alkyl groups, C6-C10 straight-chain alkyl groups, C6-C9 straight-chain alkyl groups, C6-C8 straight-chain alkyl groups, C6-C7 straight-chain alkyl groups, and C7-C9 straight-chain alkyl groups. For example, R2 is H, C6 straight-chain alkyl, C7 straight-chain alkyl, C8 straight-chain alkyl, C9 straight-chain alkyl, C10 straight-chain alkyl, C11 straight-chain alkyl, and C12 straight-chain alkyl.

[0139] In a preferred embodiment, in the compound of formula (Ⅰ) above, R1 is independently selected from a C6-C12 straight-chain alkyl group;

[0140] R2 is independently selected from C6-C12 straight-chain alkyl groups;

[0141] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, and X2 and Y2 are not simultaneously C=O or O;

[0142] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0143] n is independently selected from 2 or 3.

[0144] In a preferred embodiment, in the compound of formula (Ⅰ) above, R1 is independently selected from C7-C9 straight-chain alkyl groups;

[0145] R2 is independently selected from C7-C9 straight-chain alkyl groups;

[0146] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, and X2 and Y2 are not simultaneously C=O or O;

[0147] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0148] n is independently selected from 2 or 3.

[0149] In a preferred embodiment, in the compound of formula (Ⅰ) above, R1 is independently selected from a C6-C12 straight-chain alkyl group;

[0150] R2 is independently selected from C6-C12 straight-chain alkyl groups;

[0151] X1 and X2 are both C=O, and Y1 and Y2 are both O;

[0152] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0153] n is independently selected from 2 or 3.

[0154] In a preferred embodiment, in the compound of formula (Ⅰ) above, R1 is independently selected from C7-C9 straight-chain alkyl groups;

[0155] R2 is independently selected from C7-C9 straight-chain alkyl groups;

[0156] X1 and X2 are both C=O, and Y1 and Y2 are both O;

[0157] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0158] n is independently selected from 2 or 3.

[0159] In a preferred embodiment, in the compound of formula (Ⅰ) above, R1 and R2 are both C8 straight-chain alkyl groups;

[0160] X1 and X2 are both C=O, and Y1 and Y2 are both O;

[0161] m is independently selected from 3, 4, 5, 6, 7, and 8;

[0162] n is independently selected from 2 or 3.

[0163] This application further provides a nanoparticle composition comprising a lipid component, said lipid component comprising a compound of formula (I) provided in this application or a salt thereof or an isomer thereof.

[0164] The nanoparticle composition may include, for example, lipid nanoparticles (LNPs), liposomes, lipid vesicles, and lipid complexes.

[0165] The nanoparticle composition described in this application comprises a lipid component, which includes at least one compound according to formula (I) or a salt thereof or an isomer thereof. For example, the lipid component of the nanoparticle composition may include one or more of the compounds of the present invention. The nanoparticle composition may also include a variety of other components. For example, in addition to the compounds according to formula (I), the lipid component of the nanoparticle composition may also include one or more other lipids.

[0166] The lipid component of the nanoparticle composition may include one or more PEG or PEG-modified lipids. Such substances are alternatively referred to as polyethylene glycol-modified lipids. PEG lipids are lipids modified with polyethylene glycol. PEG lipids may be selected from the non-limiting group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof. For example, PEG lipids may be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or PEG-DSPE lipids, preferably DMG-PEG2000.

[0167] The lipid component of the nanoparticle composition may include one or more structural lipids. The structural lipids may be selected from, but are not limited to, cholesterol, coprosterol, sitosterol, ergosterol, stigmasterol, and mixtures thereof. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid includes cholesterol and corticosteroids (e.g., prednisolone, dexamethasone, prednisone, and hydrocortisone), or combinations thereof.

[0168] The lipid component of the nanoparticle composition may include one or more phospholipids, and the phospholipids used in the nanoparticle composition and method may be selected from...

[0169] Dilauroyl lecithin (DLPC)

[0170] Dimyristic phosphatidylcholine (DMPC)

[0171] Dioleoyl lecithin (DOPC)

[0172] Dipalmitoylphosphatidylcholine (DPPC)

[0173] Distearate phosphatidylcholine (DSPC)

[0174] Dioleoylphosphatidylcholine (DUPC)

[0175] Palmitoyl oleoyl phosphatidylcholine (POPC)

[0176] 1,2-Di-O-octadecyl-sn-glycerol-3-phosphate choline (18:0 Diether PC)

[0177] 1-Oleoyl-2-cholesterol dimethylsuccinate-sn-glycerol-3-phosphate choline (OChemsPC)

[0178] l-Hexadecyl-sn-glycerol-3-phosphate choline (C16 Lyso PC)

[0179] 1,2-Divinyl-sn-glycerol-3-phosphate choline,

[0180] 1,2-Diarylac-sn-glycerol-3-phosphocholine,

[0181] 1,2-Dioleoyl-SN-glycerol-3-phosphoethanolamine (DOPE)

[0182] 1,2-Distearate-sn-glycerol-3-phosphate ethanolamine,

[0183] 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine,

[0184] 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine,

[0185] 1,2-Diaryl-sn-glycerol-3-phosphate ethanolamine

[0186] 1,2-Dithiohexaenoic acid-sn-glycerol-3-phosphate ethanolamine,

[0187] 1,2-Dioleoyl-sn-glycerol-3-phosphate-(1-glycerol)sodium salt (DOPG) or sphingomyelin,

[0188] In some embodiments, the nanoparticle composition comprises DSPC. In some embodiments, the nanoparticle composition comprises DOPE. In some embodiments, the nanoparticle composition comprises both DSPC and DOPE.

[0189] In some embodiments, the LNP comprises ionizable lipids, phospholipids, cholesterol, and PEG lipids, wherein the content of ionizable lipids is 30 mol%-65 mol%, the content of the sum of phospholipids and cholesterol is 30 mol%-65 mol%, and the content of PEG lipids is 0.5 mol%-5 mol.

[0190] The nanoparticle composition may contain one or more therapeutic agents and / or preventive agents selected from vaccines or compounds capable of inducing an immune response, nucleic acids, preferably RNA, and the RNA selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

[0191] This application provides methods for delivering therapeutic and / or preventive agents to mammalian cells or organs, generating target peptides in mammalian cells, and treating diseases or conditions in mammals in need of such agents, the methods comprising administering the drug to the mammal and / or contacting mammalian cells with a composition comprising therapeutic and / or preventive nanoparticles.

[0192] In some embodiments, the therapeutic and / or preventative agent is mRNA. The mRNA may encode any target polypeptide, including any naturally occurring or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA may have any size and may possess any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA may have a therapeutic effect when expressed in cells.

[0193] The lipid component of the nanoparticle composition may include, for example, compounds according to formula (I), phospholipids (e.g., unsaturated lipids, such as DOPE or DSPC), PEG lipids, and structural lipids.

[0194] The nanoparticle composition can be characterized by a variety of methods. For example, microscopy (e.g., transmission electron microscopy or scanning electron microscopy) can be used to examine the morphology and size distribution of the nanoparticle composition. Dynamic light scattering or potentiometry (e.g., potentiometric titration) can be used to measure the zeta potential. Dynamic light scattering can also be used to determine particle size.

[0195] The average particle size of the nanoparticle composition is 50 nm to 110 nm.

[0196] The nanoparticle composition can be relatively homogeneous. A polydispersity index can be used to represent the homogeneity of the nanoparticle composition, for example, the particle size distribution of the nanoparticle composition. A small polydispersity index typically indicates a narrow particle size distribution.

[0197] This application further provides a method for specifically delivering therapeutic and / or prophylactic agents to mammalian organs, the method comprising administering the nanoparticle composition of any of the foregoing to a mammal, the administration comprising contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the organ. Therapeutic and / or prophylactic agents, such as proteins, cytotoxic agents, radioactive ions, chemotherapeutic agents, or nucleic acids (e.g., RNA, such as mRNA), may be delivered to cells or organs. In the case where the therapeutic and / or prophylactic agent is mRNA, when cells are contacted with the nanoparticle composition, the translatable mRNA may be translated in the cells to produce a target polypeptide. However, substantially untranslatable mRNAs may also be delivered to cells. Substantially untranslatable mRNAs can be used as vaccines and / or as translational components that can isolate cells to reduce the expression of other species within the cell.

[0198] In some embodiments, the nanoparticle composition can target specific types or classes of cells (e.g., cells of a specific organ or system). For example, a nanoparticle composition containing a targeted treatment and / or prevention can be specifically delivered to the lungs or spleen of a mammal. Specific delivery to a specific class of cells, organs, or systems or groups of cells means that a higher proportion of the nanoparticle composition, including treatment and / or prevention agents, is delivered to the target destination relative to other destinations. Example

[0199] Example 1: Synthesis of Compound 1

[0200]

[0201] The synthesis route is shown below:

[0202]

[0203] Specifically, compound B (126.19 g, 493.03 mmol) and DMAP (3.04 g, 22.41 mmol) were added to a solution of compound A (100.04 g, 448.21 mmol) in dichloromethane (400 ml). The mixture was stirred at room temperature until homogeneous, and then a solution of DCC (102.63 g, 493.03 mmol) in dichloromethane (100 ml) was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature for 3 hours, and the reaction was monitored by TLC. After the reaction was complete, the reaction solution was filtered, and the filter cake was washed with ethyl acetate (400 ml × 2). The filtrate was concentrated and subjected to silica gel column chromatography (ethyl acetate: petroleum ether = 0~5%) to obtain compound C (160.43 g, 77.53%).

[0204] Compound C (6.70 g, 14.51 mmol) and potassium carbonate (1.60 g, 11.61 mmol) were added to a 50 mL solution of compound D (0.50 g, 2.90 mmol) in acetonitrile. The mixture was heated to 70 °C and stirred overnight, with TLC monitoring of the reaction. After the reaction was complete, the reaction solution was filtered, and the filter cake was washed with ethyl acetate (40 mL × 2). The filtrate was collected, washed with water (100 mL × 2), washed with saturated brine (100 mL), and dried over anhydrous sodium sulfate. The solution was filtered, concentrated, and subjected to silica gel column chromatography (methanol:dichloromethane = 0–5%) to give compound 1 (0.47 g, 9.55%).

[0205] 1H NMR (600 MHz, CDCl3) δ 4.88 – 4.81 (m, 4H), 3.03 – 2.54 (m, 24H), 2.27 (t,J= 8.0 Hz, 8H), 1.65 – 1.56 (m, 8H), 1.55 – 1.47 (m, 16H), 1.37 – 1.19 (m, 128H), 0.87 (t,J= 4.0 Hz, 24H). MS-ESI (m / z): 848.4(M / 2+H) + .

[0206] Example 2 Synthesis of Compound 2

[0207]

[0208] The preparation process is the same as that of compound 1, except that 6-bromohexanoic acid and 1-decyl alcohol are used as raw materials to replace compounds A and B in the synthesis process of compound 1.

[0209] 1 H NMR (600 MHz, CDCl3) δ 4.02 (t,J= 4.0 Hz, 8H), 2.87 (brs, 16H), 2.72 – 2.64 (m, 8H), 2.28 (t,J= 8.0 Hz, 8H), 1.66 – 1.55 (m, 16H), 1.52 – 1.19 (m, 72H), 0.84 (t,J= 8.0 Hz, 12H). MS-ESI (m / z): 1190.1(M+H) + .

[0210] Example 3 Synthesis of Compound 3

[0211]

[0212] The preparation process is the same as that of compound 1, except that 6-bromohexanoic acid, 1-decyl alcohol and 1,4,7-triazacyclononane are used as raw materials to replace compounds A, B and D in the synthesis of compound 1.

[0213] 1H NMR (600 MHz, CDCl3) δ 4.02 (t,J= 4.0 Hz, 6H), 3.04 (brs, 12H), 2.76 – 2.64 (m, 6H), 2.28 (t,J= 8.0 Hz, 8H), 1.65 – 1.51 (m, 12H), 1.52 – 1.16 (m, 54H), 0.84 (t,J= 8.0 Hz, 12H). MS-ESI (m / z): 892.8(M+H) + .

[0214] Example 4 Synthesis of Compound 4

[0215]

[0216] The preparation process is the same as that of compound 1, except that 1,4,7-triazacyclononane is used as a raw material to replace compound D in the synthesis process of compound 1.

[0217] 1 H NMR (600 MHz, CDCl3) δ 4.89 (t,J= 4.0 Hz, 3H), 2.92 (brs, 12H), 2.75 – 2.66 (m, 6H), 2.31 (t,J= 8.0 Hz, 6H), 1.68 – 1.60 (m, 6H), 1.57 – 1.48(m, 12H), 1.41 – 1.22(m, 96H), 0.91 (t,J= 8.0 Hz, 18H). MS-ESI (m / z): 1271.3(M+H) + .

[0218] Example 5 Synthesis of Compound 5

[0219]

[0220] The preparation process is the same as that of compound 1, except that 4-bromobutyric acid is used as a raw material to replace compound A in the synthesis process of compound 1.

[0221] 1H NMR (600 MHz, CDCl3) δ 4.88 – 4.81 (m, 4H), 2.95 (brs, 16H), 2.78 – 2.66 (m, 8H), 2.30 (t,J= 8.0 Hz, 8H), 1.85 – 1.72 (m, 8H), 1.58 – 1.47 (m, 16H), 1.27 – 1.19 (m, 96H), 0.87 (t,J= 8.0 Hz, 24H). MS-ESI (m / z): 736.3(M / 2+H) + .

[0222] Example 6 Synthesis of Compound 6

[0223]

[0224] The preparation process is the same as that of compound 1, except that 5-bromopentanoic acid is used as a raw material to replace compound A in the synthesis process of compound 1.

[0225] 1 H NMR (600 MHz, CDCl3) δ 4.92 – 4.84 (m, 4H), 2.94 (brs, 16H), 2.78 – 2.73 (m, 8H), 2.36 (t,J= 8.0 Hz, 8H), 1.85 – 1.72 (m, 8H), 1.38 – 1.24 (m, 24H), 1.27 – 1.19 (m, 96H), 0.91 (t,J= 8.0 Hz, 24H). MS-ESI (m / z): 763.9(M / 2+H) + .

[0226] Example 7 Synthesis of Compound 7

[0227]

[0228] The preparation process is the same as that of compound 1, except that 6-bromohexanoic acid is used as a raw material to replace compound A in the synthesis process of compound 1.

[0229] 1H NMR (600 MHz, CDCl3) δ 4.88 – 4.81 (m, 4H), 2.88 (brs, 16H), 2.76 – 2.67 (m, 8H), 2.29 (t,J= 8.0 Hz, 8H), 1.85 – 1.72 (m, 8H), 1.67 – 1.48 (m, 24H), 1.37 – 1.25 (m, 112H), 0.87 (t,J= 8.0 Hz, 24H). MS-ESI (m / z): 791.9(M / 2+H) + .

[0230] Test case

[0231] Experimental Example 1: Lipid Nanoparticle (LNP) Encapsulation

[0232] The mRNA stock solution was dispersed in 20 mM acetic acid solution (pH=5.3) to a final concentration of 200 µg / mL (aqueous phase). Following the example, the compounds were mixed in a molar ratio of 40:48.5:10:1.5 (oil phase) to form a lipid mixture. The flow rates of the aqueous and oil phases were controlled using a T-junction method to mix the mRNA with the lipid mixture, resulting in LNP-encapsulated mRNA. The encapsulated LNP was diluted with buffer, then concentrated by ultrafiltration, and the diluent was replaced until the LNP concentration reached 100 µg / mL. The pH of the LNP was adjusted to approximately 7.0-8.5. Finally, the total and free mRNA content in the LNP was detected using a Ribogreen kit and 10% OTG as a demulsifier, and the encapsulation efficiency of the LNP was calculated. The final LNP product was diluted with diluent and added to a particle size distribution chamber of 1 ml. The mixture was then placed on a Malvern ZetaSizer instrument to measure the particle size of the LNP. The results are shown in Table 1.

[0233] Particle size, PDI, and encapsulation efficiency are all important quality attributes of lipid nanoparticles. As shown in the table below, the tested compounds all exhibit good encapsulation efficiency, suitable particle size for mRNA delivery, and narrow PDI.

[0234] Table 1 Characterization data of LNPs of the compounds

[0235]

[0236] Experiment 2: Testing the delivery efficiency of luciferase

[0237] 1. The mRNA expressing Luciferase was encapsulated into the LNP formulations of compounds 1, 2, 3, and 4. The LNP formulation preparation method and the mRNA encapsulation method are as described in Experimental Example 1.

[0238] 2. The LNP formulation was intravenously injected into Balb / c mice. Each mouse was injected with 0.2 ml of the LNP formulation, and the injection dose was 20 μg / mouse. The PBS control group was injected with the same volume of PBS buffer. There were 5 mice in each group.

[0239] 3. The fluorescence expression intensity of Luciferase in each mouse was detected at 6 hours.

[0240] 4. Fluorescence expression intensity detection: 10 minutes before the detection, D-fluorescein sodium salt (dosage: 150mg / kg) was injected into each mouse intraperitoneally. Then, the mice were anesthetized with isoflurane and placed in the IVIS instrument for bioluminescence detection.

[0241] 5. The delivery effect of the compound is judged by the fluorescence intensity results.

[0242] In the compound 2 group, four mice died within 3 minutes of administration, and a fifth mouse died 15 minutes later. The anatomical diagram (Figure 2) showed that the mice's lungs were congested and their stomachs were bubbly and contained fluid. It is evident that compound 2 has significant toxicity.

[0243] Based on the total fluorescence data 6 hours after intravenous administration (Figure 1), it can be seen that both compound 1 and compound 4 groups can effectively express Luciferase and achieve effective mRNA delivery, while compound 3 group is basically not expressed and has poor delivery effect.

[0244] Fluorescence expression detection of compound 1 in various organs showed that the fluorescence expression was mainly concentrated in the lungs (55.69%), followed by the spleen (37.59%), accounting for more than 93% of the total fluorescence value. The heart, liver, kidneys, stomach, and intestines showed very little expression, accounting for only 0.10%, 3.79%, 0.80%, 0.40%, and 1.64% of the total fluorescence value, respectively. This indicates that LNP prepared using compound 1 can achieve targeted delivery of mRNA to the lungs and spleen.

[0245] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, ； Formula (I) in, R1 is independently selected from C1-C14 alkyl groups; R2 is independently selected from hydrogen or C1-C14 alkyl groups; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, and X2 and Y2 are not simultaneously C=O or O; m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; n is independently selected from 1, 2, 3, 4, or 5.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is independently selected from 1, 2, 3, 4, preferably 2 or 3.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein m is independently selected from 2, 3, 4, 5, 6, 7 or 8, preferably 3, 4, 5, 6 or 7.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R1 is independently selected from a straight-chain alkyl group of C6-C12, preferably a straight-chain alkyl group of C7-C9; R2 is independently selected from hydrogen or a C6-C12 straight-chain alkyl group, preferably a C7-C9 straight-chain alkyl group.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein R1 is independently selected from a straight-chain alkyl group of C6-C12, preferably a straight-chain alkyl group of C7-C9; R2 is independently selected from C6-C12 straight-chain alkyl groups, preferably C7-C9 straight-chain alkyl groups; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, X2 and Y2 are not simultaneously C=O or O. Preferably, X1 and X2 are both C=O, and Y1 and Y2 are both O. m is independently selected from 3, 4, 5, 6, 7, and 8; n is independently selected from 2 or 3.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, The compound in (I) is selected from Compound 1: ， Compound 2: ， Compound 3: ， Compound 4: ， Compound 5: ， Compound 6: or Compound 7: 。 7. A lipid nanoparticle composition comprising a lipid component, said lipid component comprising any one of claims 1-6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

8. The lipid nanoparticle composition according to claim 7, wherein the lipid component further comprises phospholipids, structural lipids, and / or PEG lipids; The phospholipids mentioned above are preferably selected from one or more of the following compounds: Dilauroyl lecithin (DLPC) Dimyristic phosphatidylcholine (DMPC) Dioleoyl lecithin (DOPC) Dipalmitoylphosphatidylcholine (DPPC) Distearate phosphatidylcholine (DSPC) Dioleoylphosphatidylcholine (DUPC) Palmitoyl oleoyl phosphatidylcholine (POPC) 1,2-Di-O-octadecyl-sn-glycerol-3-phosphate choline (18:0 Diether PC) 1-Oleoyl-2-cholesterol dimethylsuccinate-sn-glycerol-3-phosphate choline (OChemsPC) l-Hexadecyl-sn-glycerol-3-phosphate choline (C16 Lyso PC) 1,2-Divinyl-sn-glycerol-3-phosphate choline, 1,2-Diarylac-sn-glycerol-3-phosphocholine, 1,2-Dioleoyl-SN-glycerol-3-phosphoethanolamine (DOPE) 1,2-Distearate-sn-glycerol-3-phosphate ethanolamine, 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine, 1,2-Divinyl-sn-glycerol-3-phosphate ethanolamine, 1,2-Diaryl-sn-glycerol-3-phosphate ethanolamine 1,2-Dithiohexaenoic acid-sn-glycerol-3-phosphate ethanolamine, 1,2-Dioleoyl-sn-glycerol-3-phosphate-(1-glycerol)sodium salt (DOPG) or sphingomyelin, For example, the phospholipid is DOPE or DSPC; The structural lipid is preferably selected from one or more of cholesterol, coccosterol, sitosterol, ergosterol, and stigmasterol; for example, the structural lipid is cholesterol. The PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, or PEG-modified dialkylglycerol, with DMG-PEG2000 being the most preferred.

9. The nanoparticle composition according to claim 7 or 8, further comprising a therapeutic agent and / or a preventive agent, said therapeutic agent and / or preventive agent being selected from vaccines or compounds capable of inducing an immune response, nucleic acids; Preferably, the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

10. A pharmaceutical composition comprising the nanoparticle composition of any one of claims 7-9 and a pharmaceutically acceptable carrier.

11. A method of delivering a therapeutic and / or preventive agent to a mammalian organ, the method comprising administering to a subject a nanoparticle composition of any one of claims 7-9 or a pharmaceutical composition of claim 10, the administration comprising contacting an organ with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and / or preventive agent to the organ, preferably the lung and spleen.

Images