Lipid compounds and their use

Abstract

The present application provides a kind of lipid compound and its application, specifically relates to a kind of lipid compound for delivering therapeutic agent, the lipid carrier comprising it, nucleic acid lipid nanoparticle composition, pharmaceutical preparation and related application, it is compound with structural formula I or its pharmaceutically acceptable form.The lipid compound can be used in combination with other lipid components, such as neutral lipid, cholesterol, polymer-bound lipid, to form lipid nanoparticles for delivering therapeutic agent (such as nucleic acid molecule) to achieve therapeutic or preventive purposes (such as vaccination), enrich the category of ionizable lipid compounds.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of drug delivery, and particularly relates to a lipid compound and application thereof, in particular to a lipid compound for delivering a therapeutic agent, a lipid carrier comprising the same, a nucleic acid lipid nanoparticle composition, a pharmaceutical preparation and related applications. BACKGROUND

[0002] Gene therapy technology is a hot spot in the field of modern biological medicine, and nucleic acid drugs can be used to effectively prevent and treat cancer, prevent and treat bacterial and viral infections, and treat diseases with genetic causes. Due to the characteristics of nucleic acid drugs such as easy degradation and difficulty in entering cells, it is usually necessary to encapsulate them with carriers for delivery to target cells. Therefore, the development of safe and efficient delivery carriers is a prerequisite for the clinical application of gene therapy.

[0003] Lipid nanoparticles (LNP) are currently a research hotspot in the field of non-viral gene carriers. In 2018, the U.S. Food and Drug Administration (FDA) approved LNP to deliver patisiran (trade name Onpattro) to treat hereditary transthyretin amyloidosis. Since then, the use of LNP technology to deliver nucleic acid drugs has shown explosive growth. In particular, at the end of 2020, the FDA approved the COVID-19 vaccines of Moderna and BioNtech & Pfizer, both of which use LNP technology to deliver mRNA drugs, thereby achieving the prevention of SARS-CoV-2 virus.

[0004] LNP is usually composed of four lipid compounds, namely ionizable lipid, neutral lipid, steroid and polymer-bound lipid. Among them, the choice of ionizable lipid has the greatest impact on LNP. Current nucleic acid therapeutic drugs still face some challenges, mainly including insufficient delivery efficiency, low cell permeability and high sensitivity to degradation of certain nucleic acid molecules including RNA. Therefore, there is still a need to develop new lipid compounds to promote the in vitro or in vivo delivery of nucleic acid molecules to achieve therapeutic and / or prophylactic purposes. SUMMARY

[0005] In response to the deficiencies of the prior art, one of the objects of the present invention is to provide a compound or a pharmaceutically acceptable form thereof (such as a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug, etc.), which, as a lipid compound, can be used together with other lipid compounds (such as neutral lipids, charged lipids, steroids, polymer-bound lipids, etc.) to prepare lipid nanoparticles for the delivery of therapeutic or preventive agents (such as various nucleic acid molecules, specifically including mRNA), which can improve the delivery efficiency of nucleic acid drugs in the body, and lipid compounds with specific structures can be selected as lipid carriers according to the organs where the nucleic acid drugs need to be enriched.

[0006] Another object of the present invention is to provide a method for preparing the compound or a pharmaceutically acceptable form thereof.

[0007] Another object of the present invention is to provide a lipid carrier comprising the compound or a pharmaceutically acceptable form thereof.

[0008] Another object of the present invention is to provide a nucleic acid lipid nanoparticle composition comprising the compound or a pharmaceutically acceptable form thereof or the lipid carrier.

[0009] Another object of the present invention is to provide a pharmaceutical preparation comprising the compound or a pharmaceutically acceptable form thereof, or the lipid carrier, or the nucleic acid lipid nanoparticle composition.

[0010] Another object of the present invention is to provide the use of the compound or its pharmaceutically acceptable form, or the lipid carrier, or the pharmaceutical preparation of the nucleic acid lipid nanoparticle composition in the preparation of nucleic acid drugs, gene vaccines, small molecule drugs, polypeptides or protein drugs.

[0011] In order to achieve the aforementioned object of the invention, the present invention adopts the following technical solutions:

[0012] <First Aspect>

[0013] The present invention provides a compound having a structure shown in Formula I or a pharmaceutically acceptable form thereof,

[0014]

[0015] In Formula I, the dotted line represents an optional chemical bond; that is, the chemical bond shown by the dotted line exists or does not exist; when the chemical bond shown by the dotted line exists, it represents R 1 With R 2 The ring Cy is connected by chemical bonds; the ring Cy can be understood as R 1 、R 2 and N atoms together form an N heterocycle; when the chemical bond shown by the dotted line does not exist, it represents R 1 With R 2 Not connected.

[0016] R 1 、R 2 Each independently selected from C 1-10 Alkyl, or R 1 With R 2 The ring Cy is connected by a chemical bond, and the ring Cy is selected from a 3-10 membered heterocycle.

[0017] G 1 Selected from C 1-10 Alkylene.

[0018] R 3 、R 5 、R 7 Each independently selected from C 1-20 Alkylene or C 2-20 Alkenylene.

[0019] R 4 、R 6 are each independently selected from hydrogen, C 1-30 Alkyl, C 2-30 Alkenyl, C containing 1-2 heteroatoms 1-30 Alkyl, C containing 1-2 heteroatoms 2-30 Alkenyl.

[0020] R 8 Selected from C 1-30 Alkyl, C 2-30 Alkenyl, C containing 1-2 heteroatoms 1-30 Alkyl, C containing 1-2 heteroatoms 2-30 Alkenyl.

[0021] L 1 , L 2 Each independently selected from The wavy line represents the attachment site of the group.

[0022] n 1 、n 2 are each independently selected from 0 or 1, and n 1 +n 2 ≥1; that is, n 1 is 0, n 2 is 1; or, n 1 is 1, n 2 is 0; or, n 1 is 1, n 2 is 1.

[0023] When n 1 When it is 0, it means L 1 Does not exist, that is, R 3 With R 4 Directly connected by a single bond; when n 1When it is 1, R 3 , L 1 With R 4 form

[0024] Similarly, when n 2 When it is 0, it means L 2 Does not exist, that is, R 5 With R 6 Directly connected by a single bond; when n 2 When it is 1, R 5 , L 2 With R 6 form

[0025] According to a specific embodiment of the present invention, n 1 +n 2 =1.

[0026] According to a specific embodiment of the present invention, the compound has a structure shown in Formula II-1 or Formula II-2:

[0027]

[0028] Among them, R 1 、R 2 , G 1 、R 3 、R 5 、R 7 、R 4 、R 6 、R 8 has the same limitations as in Formula I.

[0029] According to a specific embodiment of the present invention, the R 1 、R 2 Each independently selected from C 1-6 Alkyl, or R 1 With R 2 They are connected by chemical bonds to form 3-6 membered heterocyclic rings.

[0030] According to a specific embodiment of the present invention, the R 1 、R 2 Each is independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, tert-butyl, n-pentyl, isopentyl, tert-pentyl, neopentyl or hexyl.

[0031] According to a specific embodiment of the present invention, the R 1 With R 2 Not connected.

[0032] According to another specific embodiment of the present invention, the R 1 With R2 They are connected by chemical bonds to form three-membered rings, four-membered rings, five-membered rings or six-membered rings.

[0033] According to a specific embodiment of the present invention, the R 1 、R 2 Each independently selected from C 1-4 Alkyl, or R 1 With R 2 Connected by chemical bonds The wavy line represents the group and G 1 connection site.

[0034] According to a preferred embodiment of the present invention, the R 1 、R 2 Each independently selected from methyl or ethyl, or, R 1 With R 2 Connected by chemical bonds

[0035] According to a specific embodiment of the present invention, the G 1 Selected from C 2-6 Alkylene, more preferably where m G An integer selected from 2-6, for example, 2, 3, 4, 5 or 6; that is, the G 1 Selected from C 2-6 Straight chain alkylene.

[0036] According to a specific embodiment of the present invention, the G 1 Selected from The wavy line represents the attachment site of the group.

[0037] According to a specific embodiment of the present invention, the R 3 、R 5 、R 7 Each independently selected from C 2-16 Alkylene, more preferably C 2-12 Alkylene, more preferably C 2-10 Alkylene.

[0038] According to a specific embodiment of the present invention, the R 3 、R 5 、R 7 Each independently selected from where m R An integer selected from 2-12, for example, 3, 4, 5, 6, 7, 8, 9, 10 or 11, etc.

[0039] According to a specific embodiment of the present invention, the R 3 、R 5 、R 7Each independently selected from The wavy line represents the attachment site of the group.

[0040] According to a specific embodiment of the present invention, the R 3 、R 7 Each independently selected from

[0041]

[0042] Preferably, the R 5 Selected from

[0043] According to a specific embodiment of the present invention, the R 4 、R 6 are each independently selected from hydrogen, C 6-22 Alkyl, C 6-22 Alkenyl, C containing 1-2 heteroatoms 6-22 Alkyl, C containing 1-2 heteroatoms 6-22 Alkenyl.

[0044] According to a preferred embodiment of the present invention, the R 4 、R 6 are each independently selected from hydrogen, C 6-22 Straight-chain or branched alkyl, C 6-22 Straight or branched alkenyl, C containing 1-2 heteroatoms 6-22 Straight or branched alkyl, C containing 1-2 heteroatoms 6-22 Straight-chain or branched alkenyl.

[0045] According to a preferred embodiment of the present invention, the C containing 1-2 heteroatoms 6-22 The heteroatom in the straight or branched alkyl group is O; and / or the C 6-22 The heteroatom in the straight-chain or branched alkenyl group is O.

[0046] According to a preferred embodiment of the present invention, the R 4 、R 6 are each independently selected from hydrogen, C 6-22 Branched alkyl, C 6-22 Branched alkenyl, C containing 1-2 heteroatoms 6-22 Branched alkyl, C containing 1-2 heteroatoms 6-22 Branched alkenyl.

[0047] According to a specific embodiment of the present invention, the R 4 、R 6 are each independently selected from hydrogen,

[0048]

[0049]

[0050] Wavy line represents the point of attachment of the group.

[0051] According to a specific embodiment of the application, when said n 1 is 0, said R 4 is hydrogen. When said n 1 is 1, said R 4 is selected from

[0052] According to a specific embodiment of the application, when said n 2 is 0, said R 6 is hydrogen. When said n 2 is 1, said R 6 is selected from

[0053] According to a specific embodiment of the application, in formula II-1, said R 4 is selected from

[0054] said R 6 is hydrogen.

[0055] According to a specific embodiment of the application, in formula II-2, said R 4 is hydrogen; said R 6 is selected from

[0056]

[0057] According to a specific embodiment of the application, said R 8 is selected from C 6-22 alkyl, C 6-22 alkenyl, C 6-22 alkyl containing 1-2 heteroatoms, C 6-22 alkenyl containing 1-2 heteroatoms.

[0058] According to a specific embodiment of the application, said R 8 is selected from C 6-22 linear or branched alkyl, C 6-22 linear or branched alkenyl, C 6-22 linear or branched alkyl containing 1-2 heteroatoms, C 6-22 linear or branched alkenyl containing 1-2 heteroatoms.

[0059] According to a preferred embodiment of the present invention, the C containing 1-2 heteroatoms 6-22 The heteroatom in the straight or branched alkyl group is O; and / or the C 6-22 The heteroatom in the straight-chain or branched alkenyl group is O.

[0060] According to a specific embodiment of the present invention, the R 8 Selected from C 6-22 Branched alkyl, C 6-22 Branched alkenyl, C containing 1-2 heteroatoms 6-22 Branched alkyl, C containing 1-2 heteroatoms 6-22 Branched alkenyl, more preferably C 6-22 Branched chain alkyl.

[0061] According to a specific embodiment of the present invention, the R 8 Selected from

[0062] Further optimization

[0063]

[0064] According to a specific embodiment of the present invention, the compound is selected from any one or a combination of at least two of Compounds 1 to 5:

[0065] Compound 1

[0066] Compound 2

[0067] Compound 3

[0068] Compound 4 Compound 5

[0069] According to a specific embodiment of the present invention, the pharmaceutically acceptable form is selected from a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug.

[0070] <Second Aspect>

[0071] The present invention provides an intermediate having a structure shown in Formula III, Formula IV, Formula V, or Formula VI:

[0072]

[0073] Among them, R 1 、R 2 , G 1 、R 3 、R 5 、R 7 、R 4 、R 6 、R 8 , L 1 , L 2 、n 1 、n 2 As defined above in the present invention.

[0074] The intermediates of the present invention can be used to prepare the compounds of the present invention or their pharmaceutically acceptable forms.

[0075] <Third Aspect>

[0076] The present invention provides a method for preparing the compound according to the first aspect or a pharmaceutically acceptable form thereof, the method comprising:

[0077] An isonitrile compound of formula III, an aldehyde compound of formula IV, an amine compound of formula V, and a carboxylic acid compound of formula VI are subjected to Ugi reaction to obtain a compound of formula I; wherein R 1 、R 2 , G 1 、R 3 、R 5 、R 7 、R 4 、R 6 、R 8 , L 1 , L 2 、n 1 、n 2 As defined above in the present invention.

[0078] <Fourth Aspect>

[0079] The present invention provides a use of the compound as described in the first aspect or a pharmaceutically acceptable form thereof in the preparation of liposome nanocarriers.

[0080] <Fifth Aspect>

[0081] The present invention provides a lipid carrier comprising the compound according to the first aspect or a pharmaceutically acceptable form thereof.

[0082] According to a specific embodiment of the present invention, the lipid carrier comprises at least one of the compound, its salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug. Such lipid carriers have high encapsulation efficiency for nucleic acid drugs, greatly improving the delivery efficiency of nucleic acid drugs in vivo.

[0083] According to a specific embodiment of the present invention, the lipid carrier comprises a combination of a first lipid compound and a second lipid compound; the first lipid compound comprises the compound as described in the first aspect or a pharmaceutically acceptable form thereof and optionally other cationic lipids, and the second lipid compound comprises any one or a combination of at least two of anionic lipids, neutral lipids, steroids, and polymer-bound lipids.

[0084] In some embodiments, the first lipid compound is the compound or a pharmaceutically acceptable form (eg, a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug) thereof.

[0085] In another embodiment, the first lipid compound is a combination of the compound or a pharmaceutically acceptable form thereof (eg, a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug) and another cationic lipid.

[0086] In some embodiments, the other cationic lipids include 1,2-dilinoleyloxy-N,N-dimethylaminopropane DLinDMA, 1,2-dioleyloxy-N,N-dimethylaminopropane DODMA, DLin-MC2-MPZ, 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane DLin-KC2-DMA, 1,2-dioleoyl-3-trimethylammonium-propane DOTAP, 1,1 Any one or a combination of at least two of '-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyl)di-dodecan-2-ol C12-200, 3β-(N-(N,'N'-dimethylaminoethane)-carbamoyl)cholesterol DC-Chol, and N-(1-(2,3-dioleoyl chloride)propyl)-N,N,N-trimethylamine chloride DOTMA.

[0087] In some embodiments, the anionic lipid includes any one or a combination of at least two of phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, dioleoylphosphatidylglycerol DOPG, 1,2-dioleoyl-sn-glycero-3-phosphatidylserine DOPS, and dimyristoylphosphatidylglycerol.

[0088] In some embodiments, the neutral lipids include at least one of 1,2-dioleoyl-sn-glycerol-3-phosphatidylethanolamine DOPE, 1,2-distearoyl-sn-glycerol-3-phosphatidylcholine DSPC, 1,2-dipalmitoyl-sn-glycerol-3-phosphatidylcholine DPPC, 1,2-dioleoyl-sn-glycerol-3-phosphatidylcholine DOPC, dipalmitoylphosphatidylglycerol DPPG, oleoylphosphatidylcholine POPC, 1-palmitoyl-2-oleoylphosphatidylethanolamine POPE, 1,2-dipalmitoyl-sn-glycerol-3-phosphoethanolamine DPPE, 1,2-dimyristoyl-sn-glycerol-3-phosphoethanolamine DMPE, distearoylphosphatidylethanolamine DSPE, and 1-stearoyl-2-oleoylphosphatidylethanolamine SOPE, or lipids modified with anionic or cationic modifying groups. The anionic or cationic modifying groups are not limited.

[0089] In some embodiments, the steroid comprises any one or a combination of at least two of cholesterol, non-sterols, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, coproporoid, and corticosteroids.

[0090] In some embodiments, the polymer-bound lipids include 1,2-dimyristoyl-sn-glyceromethoxy-polyethylene glycol PEG-DMG, dimyristoylglycerol-polyethylene glycol PEG-c-DMG, polyethylene glycol-dimyristoylglycerol PEG-C14, PEG-1,2-dimyristoyloxypropyl-3-amine PEG-c-DMA, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino(polyethylene glycol))PEG-DSPE, PEGylated phosphatidylethanolamine PEG-PE, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, Tween-20, Tween -80, 1,2-dipalmityl-sn-glycerol-methoxypolyethylene glycol PEG-DPG, 4-O-(2',3'-di(tetradecanoyloxy)propyl-1-O-(ω-methoxy(polyethoxy)ethyl)succinate PEG-s-DMG, PEG-dialkoxypropyl PEG-DAA, mPEG2000-1,2-di-O-alkyl-sn-3-carbamoylglycerol ester PEG-c-DOMG, N-acetylgalactosamine ((R)-2,3-bis(octadecyloxy)propyl-1-(methoxypoly(ethylene glycol) 2000)propylcarbamate))GalNAc-PEG-DSG or a combination of at least two thereof.

[0091] In some embodiments, the molar ratio of the first lipid compound, the anionic lipid, the neutral lipid, the steroid and the polymer-bound lipid in the lipid carrier is (20-65): (0-20): (5-25): (25-55): (0.3-15); wherein "20-65" can be 22, 25, 28, 30, 32, 35, 38, 40, 42, 45, 48, 50, 52, 55, 58, 60, 62 or 6 4, etc.; “0-20” may specifically be 1, 3, 5, 8, 10, 12, 15 or 18, etc.; “5-25” may specifically be 6, 8, 10, 12, 15, 18, 20, 22 or 24, etc.; “25-55” may specifically be 28, 30, 32, 35, 38, 40, 42, 45, 48, 50, 52 or 54, etc.; “0.3-15” may specifically be 0.5, 0.8, 1, 3, 5, 8, 10, 12 or 14, etc. For example, the molar ratio of the first lipid compound, the anionic lipid, the neutral lipid, the steroid and the polymer-bound lipid may be 20:20:5:50:5, 30:5:25:30:10, 20:5:5:55:15, 65:0:9.7:25:0.3, etc.

[0092] In which, in the first lipid compound, the molar ratio of the compound or a pharmaceutically acceptable form thereof (such as a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug) and other cationic lipids is (1-10): (0-10); wherein, "1-10" can specifically be 2, 3, 4, 5, 6, 7, 8 or 9, etc.; "0-10" can specifically be 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8 or 9, etc.; illustratively, the molar ratio can be 1:1, 1:2, 1:5, 1:7.5, 1:10, 2:1, 5:1, 7.5:1, 10:1, etc.

[0093] In some embodiments, in the lipid carrier, the molar ratio of the first lipid compound, anionic lipid, neutral lipid, steroid and polymer-bound lipid is (20-55):(0-13):(5-25):(25-51.5):(0.5-15); wherein, in the first lipid compound, the molar ratio of any of the above compounds or pharmaceutically acceptable forms thereof (such as salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs) and other cationic lipids is (3-4):(0-5).

[0094] <Sixth Aspect>

[0095] The present invention provides a nucleic acid lipid nanoparticle composition, which comprises the compound as described in the first aspect or a pharmaceutically acceptable form thereof (such as a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug), at least one of the lipid carriers as described in the fifth aspect, and a therapeutic agent or a prophylactic agent.

[0096] According to a specific embodiment of the present invention, the therapeutic agent or preventive agent is a nucleic acid drug.

[0097] According to a specific embodiment of the present invention, the therapeutic agent or prophylactic agent comprises a DNA, antisense nucleic acid, RNA, aptamer, ribozyme, immunostimulatory nucleic acid or PNA component.

[0098] According to a specific embodiment of the present invention, the DNA comprises a plasmid.

[0099] According to a specific embodiment of the present invention, the antisense nucleic acid is an antisense oligonucleic acid.

[0100] According to a specific embodiment of the present invention, the RNA includes mRNA, rRNA, circRNA, siRNA, saRNA, tRNA, snRNA, antagomir, microRNA inhibitor, microRNA activator or shRNA.

[0101] According to a specific embodiment of the present invention, the RNA comprises modified RNA.

[0102] According to a specific embodiment of the present invention, the mRNA includes a sequence encoding an RNA-guided DNA binder, more specifically, for example, an mRNA including a Cas protein.

[0103] In some embodiments, the therapeutic or prophylactic agent (nucleic acid drug) includes a guide RNA, specifically, the guide RNA includes a gRNA nucleic acid.

[0104] In some embodiments, the therapeutic or prophylactic agent (nucleic acid drug) includes mRNA and gRNA of the Cas protein.

[0105] According to a specific embodiment of the present invention, the mRNA includes an mRNA encoding an RNA-guided nuclease or an mRNA encoding a base editor, and a gRNA.

[0106] In some embodiments, the gRNA is modified.

[0107] According to a specific embodiment of the present invention, the nuclease includes Cas9, Cas12, Cas13, IscB, TnpB, IsrB and homologs thereof.

[0108] In some embodiments, the therapeutic or prophylactic agent (nucleic acid drug) is in a mass ratio of 1 : (3-40) to the compound or its pharmaceutically acceptable form (e.g., salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug) thereof, such as 1 :3, 1 :5, 1 :8, 1 :10, 1 :12, 1 :15, 1 :18, 1 :20, 1 :22, 1 :25, 1 :28, 1 :30, 1 :32, 1 :35, or 1 :38, etc.

[0109] In some embodiments, the therapeutic or prophylactic agent (nucleic acid drug) is in a mass ratio of 1 : (3-40) to the lipid carrier, such as 1 :3, 1 :5, 1 :8, 1 :10, 1 :12, 1 :15, 1 :18, 1 :20, 1 :22, 1 :25, 1 :28, 1 :30, 1 :32, 1 :35, or 1 :38, etc.

[0110] <Seventh Aspect>

[0111] The present application provides a pharmaceutical composition comprising at least one of the compound or its pharmaceutically acceptable form (e.g., salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug) as described in the first aspect, the lipid carrier as described in the fifth aspect, the nucleic acid lipid nanoparticle composition as described in the sixth aspect, and a pharmaceutically acceptable excipient.

[0112] <Eighth Aspect>

[0113] The present application provides a pharmaceutical preparation comprising at least one of the compound or its pharmaceutically acceptable form (e.g., salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug) as described in the first aspect, the lipid carrier as described in the fifth aspect, the nucleic acid lipid nanoparticle composition as described in the sixth aspect, and a pharmaceutically acceptable excipient.

[0114] According to a specific embodiment of the present application, the pharmaceutically acceptable excipient comprises any one of or a combination of at least two of excipients, carriers, diluents, fillers, binders, wetting agents, disintegrants, emulsifiers, co-solvents, solubilizers, osmotic pressure adjusting agents, pH adjusting agents, antioxidants, buffers.

[0115] In some embodiments, the particle size of the pharmaceutical preparation is 30-500 nm, and exemplarily, the particle size can be 30 nm, 50 nm, 80 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, or 500 nm, etc.

[0116] In some embodiments, the encapsulation efficiency of the therapeutic or preventive agent (nucleic acid drug) in the pharmaceutical preparation is greater than 50%. Exemplarily, the encapsulation efficiency can be 55%, 60%, 65%, 70%, 75%, 79%, 80%, 85%, 89%, 90%, 93% or 95%, etc.

[0117] <Ninth Aspect>

[0118] The present invention provides a compound as described in the first aspect or a pharmaceutically acceptable form thereof (such as a salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug), a lipid carrier as described in the fifth aspect, a nucleic acid lipid nanoparticle composition as described in the sixth aspect, a pharmaceutical composition as described in the seventh aspect, and a pharmaceutical preparation as described in the eighth aspect for use in the preparation of nucleic acid drugs, gene vaccines, small molecule drugs, polypeptides or protein drugs.

[0119] In some embodiments, the nucleic acid lipid nanoparticle composition or the pharmaceutical formulation is used to treat or prevent a disease or disorder in a subject in need of treatment or prevention.

[0120] In some embodiments, the subject is a mammal.

[0121] In some embodiments, the subject is a human.

[0122] In some embodiments, the disease or condition is selected from a metabolic disease, a genetic disease, cancer, a cardiovascular disease, and an infectious disease.

[0123] In some embodiments, the metabolic disease comprises familial hypercholesterolemia (FH).

[0124] In some embodiments, the genetic disease comprises transthyretin amyloidosis (ATTR), primary hyperoxaluria (PH1), or hereditary angioedema (HAE).

[0125] In some embodiments, the infectious disease comprises hepatitis B.

[0126] The present invention also provides a method for preventing, improving or treating a disease or condition in a subject in need thereof, the method comprising administering to the subject the nucleic acid lipid nanoparticle composition as described in the sixth aspect, the pharmaceutical composition as described in the seventh aspect, or the pharmaceutical preparation as described in the eighth aspect.

[0127] The present invention also provides a method for delivering a therapeutic agent or a prophylactic agent to a subject's cells, the method comprising administering to the subject at least one of the nucleic acid lipid nanoparticle composition as described in the sixth aspect, the pharmaceutical composition as described in the seventh aspect, and the pharmaceutical preparation as described in the eighth aspect, wherein the administration comprises contacting the subject's cells with the nucleic acid lipid nanoparticle composition, pharmaceutical composition, or pharmaceutical preparation, thereby delivering the therapeutic agent or prophylactic agent to the subject's cells.

[0128] In some embodiments, the routes of administration include oral, intranasal, intravenous, intraperitoneal, intramuscular, intraarticular, intralesional, intratracheal, subcutaneous, and intradermal.

[0129] In some embodiments, the nucleic acid lipid nanoparticle composition, pharmaceutical composition, or pharmaceutical formulation is administered, for example, via an enteral or parenteral route of administration.

[0130] In some embodiments, a dose of about 0.001 mg / kg to about 10 mg / kg of the nucleic acid lipid nanoparticle composition, pharmaceutical composition, or pharmaceutical formulation is administered to the subject.

[0131] The present invention also provides a method for producing a target protein or target polypeptide in a subject cell, the method comprising contacting the subject cell with the nucleic acid lipid nanoparticle composition described in the sixth aspect, wherein the therapeutic agent or preventive agent is mRNA, and the mRNA encodes the target protein or target polypeptide, whereby the mRNA can be translated in the cell to produce the target protein or target polypeptide.

[0132] Compared with the prior art, the present invention has the following beneficial effects:

[0133] The present invention provides a series of novel structural compounds of formula I. These compounds can be used as ionizable lipids and, together with other lipid compounds, prepare lipid carriers. These lipid carriers have controllable particle size, uniform distribution, and high encapsulation efficiency. The synthesis method is simple, has high yield, can be rapidly synthesized, and is low in cost. The compounds of the present invention can be used to deliver nucleic acid drugs, gene vaccines, small molecule drugs, polypeptides, or protein drugs, enriching the variety of ionizable lipid compounds and improving the delivery efficiency of nucleic acid drugs in vivo. BRIEF DESCRIPTION OF THE DRAWINGS

[0134] Figure 1 Schematic diagram of the delivery strategy for detecting PCSK9 gene editing efficiency in mouse liver cells in one embodiment of the present invention;

[0135] Figure 2 A comparison of the PCSK9 gene editing efficiency of base editors encapsulated by different lipid compounds in mouse liver cells. DETAILED DESCRIPTION

[0136] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0137] For easier understanding of the present invention, certain technical and scientific terms are specifically defined below. Unless otherwise clearly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by those skilled in the art.

[0138] In this specification, a numerical range expressed using "a numerical value A - a numerical value B" means a range including the endpoints A and B.

[0139] In this specification, the use of “substantially” or “essentially” means that the standard deviation from a theoretical model or theoretical data is within a range of 5%, preferably 3%, and more preferably 1%.

[0140] In this specification, the use of "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0141] As used herein, "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event occurs and instances where it does not.

[0142] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "embodiments," etc., mean that the specific elements (e.g., features, structures, properties, and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0143] Before the present invention is further described, it is to be understood that the present invention is not limited to the particular embodiments described herein; it is to be understood that the terminology used herein is for the purpose of describing only and is not intended to be limiting of the particular embodiments.

[0144] [Definition of terms]

[0145] Unless otherwise stated, the following terms have the following meanings:

[0146] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is substantially non-toxic to living organisms. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by reacting a compound of the present invention with a pharmaceutically acceptable inorganic / organic acid or inorganic / organic base. Such salts are also referred to as acid addition salts or base addition salts. Common inorganic acids include (but are not limited to) hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.; common organic acids include (but are not limited to) trifluoroacetic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, oxalic acid, formic acid, acetic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.; common inorganic bases include (but are not limited to) sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, etc.; common organic bases include (but are not limited to) diethylamine, triethylamine, ethambutol, etc.

[0147] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that possesses at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.) resulting in a perpendicular asymmetric plane, thereby rotating plane-polarized light. Because the compounds of the present invention contain asymmetric centers and other chemical structures that may lead to stereoisomerism, the present invention also encompasses these stereoisomers and mixtures thereof. Because the compounds of the present invention and their salts contain asymmetric carbon atoms, they can exist as single stereoisomers, racemates, enantiomers, and mixtures of diastereomers. Typically, these compounds can be prepared as racemic mixtures. However, if desired, such compounds can be prepared or isolated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched in a single stereoisomer (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85%, or ≥80%). Individual stereoisomers of a compound are synthesized from optically active starting materials containing the desired chiral center, or by preparing a mixture of enantiomeric products followed by separation or resolution, for example, by conversion to a mixture of diastereomers followed by separation or recrystallization, chromatography, use of a chiral resolving agent, or direct separation of the enantiomers on a chiral chromatographic column. Starting compounds of specific stereochemistry are either commercially available or prepared as described herein and resolved by methods well known in the art.

[0148] The term "tautomer" (or "tautomeric form") refers to structural isomers that exist in equilibrium with one another through low energy barriers. If tautomerism is possible (as in solution), a chemical equilibrium of the tautomers can be reached. For example, prototropic tautomers (or proton-shift tautomers) include, but are not limited to, interconversions by proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imidol isomerization, and the like. Unless otherwise specified, all tautomeric forms of the compounds of the application are within the scope of the application.

[0149] The term "solvate" refers to a molecular complex of a compound of the application or a pharmaceutically acceptable salt thereof with one or more solvent molecules. Common solvates include, but are not limited to, hydrates, ethanol solvates, acetone solvates, and the like.

[0150] The term "chelate" refers to a complex having a cyclic structure, formed by chelation, the formation of several coordinate bonds between a single metal ion and two or more ligands.

[0151] The term "non-covalent complex" refers to a complex formed by the interaction of a compound with another molecule, wherein no covalent bond is formed between the compound and the molecule. Complexation can occur, for example, by van der Waals interactions, hydrogen bonding, and electrostatic interactions (also known as ionic bonding).

[0152] The term "prodrug" refers to a derivative of a compound of the application that is applicable to a patient and that can provide, directly or indirectly, the compound of the application. Particularly preferred derivatives or prodrugs are compounds that increase the bioavailability of the compound of the application when administered to a patient (e.g., are more readily absorbed into the blood), or that facilitate delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise specified, all prodrug forms of the compounds of the application are within the scope of the application, and various prodrug forms are well known in the art.

[0153] The term "each independently" means that at least two groups (or ring systems) present in a structure that have the same or similar range of values can have the same or different meanings in a particular instance. For example, if group X and group Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when group X is hydrogen, group Y can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl; and similarly, when group Y is hydrogen, group X can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl.

[0154] The terms "comprising" and "including," as used herein, are used in their open, non- limiting sense.

[0155] The term "alkyl" refers to a monovalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing no unsaturation, and connected to other fragments by a single bond, including (but not limited to) methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl and tert-butyl. For example, the term "C 1-30 "Alkyl" refers to a saturated monovalent straight or branched alkane group containing 1 to 30 carbon atoms, specifically C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 12 、C 14 、C 16 、C 18 、C 20 、C 22 、C 24 、C 26 or C 28 etc. straight chain or branched chain alkyl. 1-10 "Alkyl" refers to a saturated monovalent straight or branched hydrocarbon group containing 1 to 10 carbon atoms, specifically C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 A straight chain or branched chain alkyl group.

[0156] The term "alkylene" refers to a divalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing no saturation, and connected to other fragments by two single bonds, including (but not limited to) methylene, 1,1-ethylene and 1,2-ethylene. For example, "C 1-20 "Alkylene" refers to a saturated divalent straight or branched chain alkyl group containing 1 to 20 carbon atoms, specifically C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 12 、C 14 、C 16 、C 18 、C 20 etc., a straight or branched chain alkylene group.

[0157] The term "cycloalkyl" or "ring" refers to a saturated, monocyclic or polycyclic (e.g., bicyclic, tricyclic or tetracyclic) non-aromatic hydrocarbon group consisting only of carbon and hydrogen atoms. Cycloalkyl groups may include fused, bridged or spirocyclic ring systems. For example, "C 3-6 "Cycloalkyl" refers to a cycloalkyl group having 3 to 6 carbon atoms. For example, the cycloalkyl group may be cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or bicyclo[2.2.1]heptyl, etc.

[0158] The term "cycloalkylene" refers to a divalent group obtained by removing a hydrogen atom from a cycloalkyl group as defined above, including but not limited to cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and cycloheptylene. For example, "C 3-30 The "cycloalkylene group" refers to a divalent group obtained by removing a hydrogen atom from a cycloalkyl group containing 3 to 30 carbon atoms.

[0159] The term "branched alkyl" or "branched alkyl" refers to an alkane radical that is attached to a parent molecule and forms at least two branched structures. For example

[0160] The term "alkenyl" refers to a monovalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, and connected to other fragments by a single bond, including (but not limited to) vinyl, propenyl, allyl, isopropenyl, butenyl and isobutenyl groups. For example, "C 2-30 "Alkenyl" refers to a monovalent straight or branched alkene group containing 2 to 30 carbon atoms and having at least one carbon-carbon double bond (>C=C<), specifically C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 12 、C 14 、C 16 、C 18 、C 20 、C 22 、C 24 、C 26 or C 28 Straight-chain or branched alkenyl groups.

[0161] The term "alkenylene" refers to a divalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing at least one double bond, and connected to other fragments by two single bonds, including (but not limited to) vinylene, etc. For example, "C 2-20 "Alkenylene" refers to a divalent straight or branched hydrocarbon group containing 2 to 20 carbon atoms and having at least one carbon-carbon double bond (>C=C<), specifically C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 12 、C 14 、C 16 、C 18 、C 20 Straight-chain or branched alkenylene.

[0162] The term "alkynyl" refers to a monovalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon triple bond, and connected to other moieties by a single bond, including (but not limited to) ethynyl, propynyl, butynyl and pentynyl groups. For example, "C2-30 "Alkynyl" refers to a monovalent straight or branched chain hydrocarbon radical containing from 2 to 30 carbon atoms and having at least one carbon-carbon triple bond.

[0163] The term "alkynylene" refers to a divalent straight or branched alkane group consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon triple bond, and connected to other fragments by two single bonds, including (but not limited to) ethynylene. 2-30 "Alkyne" refers to a divalent straight or branched chain hydrocarbon group containing 2 to 30 carbon atoms and having at least one carbon-carbon triple bond.

[0164] The term "cycloalkenyl" refers to an unsaturated, monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) non-aromatic hydrocarbon group consisting solely of carbon and hydrogen atoms. Cycloalkenyl groups may include cyclic, bridged, or spirocyclic ring systems. Examples include cyclopropenyl and cyclobutenyl.

[0165] The term "cycloalkenylene" refers to a divalent group obtained by removing a hydrogen atom from a cycloalkenyl group as defined above, including but not limited to cyclopropenylene and cyclobutenylene. For example, "C 3-30 The "cycloalkenylene group" refers to a divalent group obtained by removing a hydrogen atom from a cycloalkenyl group containing 3 to 30 carbon atoms.

[0166] The term "branched alkenyl" or "branched alkenyl" refers to an alkene radical attached to a parent molecule and forming at least two branched structures. For example

[0167] The term "heterocyclyl" or "heterocycle" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic, for example, fused, bridged or spiro) non-aromatic group, the ring atoms of which are composed of carbon atoms and at least one heteroatom selected from N, O and S, wherein the S atom is optionally substituted to form S(=O), S(=O)2 or S(=O)(=NRx), and Rx is independently selected from H or C 1-4 Alkyl. If the valence bond requirements are met, the heterocyclic group can be connected to the rest of the molecule through any one of the ring atoms. For example, "3-10 membered heterocyclic group" or "3-10 membered heterocycle" refers to a heterocyclic ring (radical) having 3 to 10 ring atoms. For example, the heterocyclic ring (radical) can be oxirane (radical), aziridine (radical), azetidine (radical), oxetane (radical), tetrahydrofuran (radical), dioxol (radical), pyrrolidone (radical), imidazolidine (radical), pyrazolidine (radical), tetrahydropyran (radical), piperidine (radical), piperazine (radical), morpholine (radical), thiomorpholine (radical), dithiane (radical) or trithiane (radical).

[0168] The term "aryl" refers to a monocyclic or fused polycyclic aromatic hydrocarbon group having a conjugated π electron system. For example, the term "C6-10 Aryl" refers to an aromatic group having 6 to 10 carbon atoms. For example, aryl can be phenyl, naphthyl, anthryl, phenanthryl, acenaphthyl, azulenyl, azulenyI, fluorenyl, indenyl, pyrenyl, and the like.

[0169] The term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic group having a conjugated pi-electron system, whose ring atoms are composed of carbon atoms and at least one heteroatom selected from N, O, and S. If valence requirements are met, a heteroaryl group can be attached to the rest of the molecule through any one of the ring atoms. For example, the term "5-10 membered heteroaryl" refers to a heteroaryl group having 5 to 10 ring atoms. For example, heteroaryl can be thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and benzo derivatives thereof, pyrrolopyridyl, pyrrolopyrazinyl, pyrazolopyridyl, imidazopyridyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, purinyl, and the like.

[0170] The term "C 1-30 Alkyl" can be understood as a monovalent radical formed by replacing one or two -CH2- not directly connected in an alkyl group with a heteroatom, C 1-30 The definition of alkyl is as previously described and for the sake of brevity will not be repeated; the heteroatom can be O or S, and the like.

[0171] The term "C 2-30 Alkenyl" can be understood as a monovalent radical formed by replacing one or two -CH2- not directly connected in an alkenyl group with a heteroatom, C 2-30 The definition of alkenyl is as previously described and for the sake of brevity will not be repeated; the heteroatom can be O or S, and the like.

[0172] The term "halogen" refers to fluorine F, chlorine Cl, bromine Br, and iodine I.

[0173] The term "hydroxyl" refers to -OH.

[0174] The term "cyano" refers to -CN.

[0175] The term "amino" refers to -NH2.

[0176] The term "nitro" refers to -NO2.

[0177] The term "oxo" refers to (=O).

[0178] [Preparation method]

[0179] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials can be obtained commercially unless otherwise specified.

[0180] In the present invention, "appropriate amount" means that the amount of the added solvent or the amount of the drug can be adjusted in a wide range and has little effect on the synthesis result, and is not specifically limited.

[0181] In the following examples, all solvents and drugs used were of analytical or chemical purity; all solvents were redistilled before use; and all anhydrous solvents were treated according to standard methods or literature methods.

[0182] Examples

[0183] Where specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or the conditions recommended by the manufacturer. Reagents or instruments used without specifying the manufacturer are conventional products that can be obtained commercially. It will be appreciated by those skilled in the art that the examples describe the present invention by way of example and are not intended to limit the scope of protection claimed in the present invention. The technical features involved in the various embodiments of the present invention may be combined with each other as long as they do not conflict with each other. All disclosures and other references mentioned herein are incorporated herein by reference in their entirety.

[0184] Example 1

[0185] Compound 1: Synthesis of 2-butyloctyl 10-(7-butyl-10,18-dioxyylidene-19-aza-9-oxaheptacosan-19-yl)-11-((3-(diethylamino)propyl)amino)-11-oxyylideneundecanoate

[0186]

[0187] Step 1: Synthesis of compound 1-2

[0188] 3-(Diethylamino)propan-1-amine (2.00 g, 15.36 mmol, 1.0 eq) was added to 50 ml of ethyl formate (HCO₂Et) and stirred at 60°C for 5 h. The mixture was concentrated under reduced pressure to afford compound 1-2: N-(3-(diethylamino)propyl)methaneamide (1.90 g, 78.2% yield). MS: m / z [M+H] + =159.1.

[0189] Step 2: Synthesis of Compounds 1-3

[0190] Into a 100 mL round bottom flask was added compound 1-2 (2.00 g, 12.64 mmol, 1.0 eq), triethylamine (TEA, 7.67 g, 75.84 mmol, 6.0 eq) and dichloromethane (30 mL) and cooled in an ice bath. Phosphorous oxychloride (2.91 g, 18.96 mmol, 1.5 eq) was added dropwise. After the addition was complete, the reaction mixture was allowed to warm to 25 °C and stirred for 2 h. The reaction mixture was concentrated under reduced pressure and purified by column chromatography to afford compound 1-3: diethyl(3-isocyanopropyl)amine (1.20 g, 67.7% yield).

[0191] Step 3: Synthesis of compound 1-6

[0192] Into a 100 mL round bottom flask was added azelaic acid (10.10 g, 53.65 mmol, 5.0 eq), 4-dimethylpyridine (DMAP, 0.66 g, 5.37 mmol, 0.5 eq), N,N-diisopropylethylamine (DIEA, 13.87 g, 107.30 mmol, 10.0 eq), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCl, 3.09 g, 16.09 mmol, 1.5 eq) and 50 mL dichloromethane (DCM), and stirred at room temperature for half an hour. 2-Butyloctan-l-ol (2.00 g, 10.73 mmol, 1.0 eq) was added. After 16 h at room temperature, the reaction mixture was concentrated under reduced pressure to remove the solvent, diluted with 100 mL water, extracted with 100 mL ethyl acetate three times, the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to afford compound 1-6: 9-((2-butyloxy)oxy)-9-oxoazelaic acid (3.00 g, 78.4% yield).

[0193] Step 4: Synthesis of compound 1-8

[0194] Into a 100 mL flask was added 10-hydroxydecanoic acid (5.00 g, 26.56 mmol, 1.0 eq), dihydropyran (DHP, 3.35 g, 39.84 mmol, 1.5 eq), p-toluenesulfonic acid (TsOH, 0.23 g, 1.33 mmol, 0.05 eq) and tetrahydrofuran (THF, 50 mL). The reaction mixture was stirred at room temperature for 16 h, concentrated under reduced pressure, and purified by column chromatography to afford compound 1-8: 10-(3,4,5,6-tetrahydro-2H-pyran-2-yloxy)decanoic acid (6.30 g, 87.1% yield).

[0195] Step 5: Synthesis of compound 1-9

[0196] Compound 1-8 (6.30 g, 23.13 mmol, 1.0 eq), 2-butyloctan-1-ol (6.46 g, 34.70 mmol, 1.5 eq), 4-dimethylpyridine (DMAP, 1.41 g, 11.56 mmol, 0.5 eq), N, N-diisopropyl ethylamine (DIEA, 8.97 g, 69.39 mmol, 3.0 eq) were added into a round bottom flask containing 60 mL of dichloromethane, and finally EDCl (6.65 g, 34.70 mmol, 1.5 eq) was added. The reaction was carried out at room temperature for 16 h. Concentration under reduced pressure and column chromatography yielded compound 1-9: 10-(3,4,5,6-tetrahydro-2H-pyran-2-yloxy)decanoic acid-2-butyloctyl ester (5.30 g, yield 52.0%).

[0197] Step 6: Synthesis of compound 1-10

[0198] Compound 1-9 (5.30 g, 12.03 mmol, 1.0 eq) and p-toluenesulfonic acid (2.07 g, 12.03 mmol, 1.0 eq) were added into a 100 mL flask containing 20 mL of ethanol. The reaction was carried out at room temperature for 16 h. The solvent was removed by concentration under reduced pressure and column chromatography yielded compound 1-10: 10-hydroxydecanoic acid-2-butyloctyl ester (1.60 g, yield 37.3%).

[0199] Step 7: Synthesis of compound 1-11

[0200] Compound 1-10 (0.80 g, 2.24 mmol, 1.0 eq) and sodium bicarbonate (0.47 g, 5.60 mmol, 2.5 eq) were added into a round bottom flask containing 10 mL of dichloromethane, and Dess-Martin oxidizing agent (1.14 g, 2.69 mmol, 1.2 eq) was added portionwise under ice bath. After the addition was completed, the reaction was continued at 0 °C for 2 h, and saturated sodium thiosulfate solution was added to quench, and the organic phase was extracted with 30 mL of dichloromethane three times, combined, washed with saturated brine, dried over anhydrous sodium sulfate, and column chromatography yielded compound 1-11: 9-formyl nonanoic acid-2-butyloctyl ester (570.0 mg, yield 71.6%).

[0201] Step 8: Synthesis of compound 1

[0202] In a 25 ml round-bottom flask were added 5 ml of methanol, compound 1-11 (150.0 mg, 0.42 mmol, 1.0 eq), and n-octylamine (54.0 mg, 0.42 mmol, 1.0 eq) in sequence. After reacting at room temperature for 0.5 h, compound 1-6 (0.15 g, 0.42 mmol, 1.0 eq) was added. After stirring at room temperature for 0.5 h, compound 1-3 (59.0 mg, 0.42 mmol, 1.0 eq) was added. The reaction solution was stirred at room temperature for 16 h, diluted with 50 ml of water, and extracted three times with 30 ml of n-hexane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain compound 1: 2-butyloctyl 10-(7-butyl-10,18-dioxyylidene-19-aza-9-oxaheptacosane-19-yl)-11-((3-(diethylamino)propyl)amino)-11-oxyylideneundecanoate (80.2 mg, 19.7% yield). MS: m / z [M+H] + =962.9. 1 H-NMR (400MHz, CDCl3): δ6.72 (s, 1H), 4.68 (s, 1H), 3.92-3.75 (m, 4H), 3.56-3.24 (m, 4H), 2.55-2.38 (m, 4 H), 2.37-2.10(m, 10H), 2.00-1.58(m, 15H), 1.50-1.42(m, 14H), 1.40-1.12(m, 47H), 0.98-0.75(m, 15H).

[0203] Example 2

[0204] Compound 2: Synthesis of 2-butyloctyl 10-(7-butyl-10,18-dioxyylidene-19-aza-9-oxaheptacosan-19-yl)-11-((3-(dimethylamino)propyl)amino)-11-oxyylideneundecanoate

[0205]

[0206] Example 2 was synthesized according to the method of Example 1, except that 3-(diethylamino)propan-1-amine in the first step was replaced with 3-(dimethylamino)propan-1-amine. The yield was 35.1%. MS: m / z [M+H] + =934.8. 1H-NMR (400MHz, CDCl3): δ6.78 (s, 1H), 4.66 (s, 1H), 3.99-3.79 (m, 4H), 3.62-3.26 (m, 4H), 2.55-2.37 (m, 4 H), 2.33-2.14(m, 10H), 2.09-1.52(m, 15H), 1.55-1.41(m, 14H), 1.40-1.15(m, 43H), 1.05-0.78(m, 15H).

[0207] Example 3

[0208] Compound 3: Synthesis of 2-butyloctyl 10-(7-butyl-10,18-dioxyylidene-19-aza-9-oxaheptacosan-19-yl)-11-((3-(tetrahydro-1H-pyrrol-1-yl)propyl)amino)-11-oxyylideneundecanoate

[0209]

[0210] Example 3 was synthesized according to the method of Example 1, except that 3-(diethylamino)propan-1-amine in the first step was replaced with 3-(tetrahydro-1H-pyrrol-1-yl)propan-1-amine. The yield was 41.4%. MS: m / z [M+H] + =960.9. 1 H-NMR (400MHz, CDCl3): δ6.66 (s, 1H), 4.65 (s, 1H), 3.90-3.77 (m, 4H), 3.55-3.23 (m, 4H), 2.58-2.42 (m, 4H), 2.37-2.29 (m, 4H), 2.24-2.10(m, 6H), 2.08-1.79(m, 10H), 1.73-1.58(m, 5H), 1.55-1.46(m, 14H), 1.42-1.17(m, 45H), 1.01-0.77(m, 15H).

[0211] Example 4

[0212] Compound 4: Synthesis of 10-(9-((2-butyloctyl)oxy)-1,9-dioxyylidenonyl)-3-ethyl-9-octyl-8-oxyylidene-3,7,10-triazatridec-13-yl 4,4-bis(((5Z)-oct-5-enyl)oxy)butanoate

[0213]

[0214] Compound 4-1 was synthesized by a four-component Ugi reaction of diethyl(3-isocyanatopropyl)amine (Compound 1-3), n-nonanal, hydroxypropylamine, and 9-((2-butyloctyl)oxy)-9-oxyylidenenonanoic acid (Compound 1-6) according to the method of Example 1 with a yield of 42.3%. MS: m / z [M+H] + =696.6.

[0215] Step 1: Synthesis of compound 4

[0216] Compound 4-1 (500.0 mg, 0.72 mmol, 1.0 eq), compound 4-2 (0.37 g, 1.08 mmol, 1.5 eq), DMAP (88.0 mg, 0.072 mmol, 0.1 eq), EDCl (0.21 g, 1.08 mmol, 1.5 eq), DIEA (0.19 g, 1.44 mmol, 2.0 eq) were added to 3 ml of dichloromethane in sequence, and stirred at room temperature for 16 h before addition. The mixture was diluted with 20 ml of water and extracted three times with 10 ml of dichloromethane, respectively. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain compound 4: 4,4-bis(((5Z)-oct-5-enyl)oxy)butanoic acid-10-(9-((2-butyloctyl)oxy)-1,9-dioxylidenonyl)-3-ethyl-9-octyl-8-oxylide-3,7,10-triazatridecan-13-yl ester (0.30 g, 41.0% yield). MS: m / z [M+H] + =1018.9. 1 H-NMR (400MHz, CDCl3): δ7.25 (s, 1H), 5.45-5.25 (m, 4H), 4.75-4.60 (m, 1H), 4.58-4.41 (m, 1H), 4.20-3.82 (m, 4H), 3.62-3.48 (m, 2H), 3. 47-3.05 (m, 6H), 2.70-2.23 (m, 12H), 2.10-1.81 (m, 13H), 1.75-1.51 (m, 13H), 1.50-1.48 (m, 5H), 1.47-1.10 (m, 38H), 1.10-0.75 (m, 15H).

[0217] Example 5

[0218] Compound 5: Synthesis of 9-(9-((2-butyloctyl)oxy)-1,9-dioxyylidenonyl)-3-ethyl-8-octyl-7-oxyylidene-3,6,9-triazadodec-12-yl 4,4-bis(((5Z)-oct-5-enyl)oxy)butanoate

[0219]

[0220] Example 5 was synthesized according to the method of Example 4, except that 3-(diethylamino)propan-1-amine was replaced with 3-(dimethylamino)propan-1-amine. The yield was 35.1%. MS: m / z [M+H] + =1004.8. 1 H-NMR (400MHz, CDCl3): δ7.28 (s, 1H), 5.40-5.26 (m, 4H), 4.81-4.63 (m, 1H), 4.54-4.43 (m, 1H), 4.32-3.85 (m, 4H), 3.66-3.10 ( m, 8H), 2.77-2.25 (m, 12H), 2.12-1.81 (m, 13H), 1.80-1.78 (m, 13H), 1.77-1.50 (m, 5H), 1.44-1.13 (m, 36H), 0.99-0.75 (m, 15H).

[0221] Example 6

[0222] This example provides the preparation, characterization, and in vivo editing experimental evaluation of lipid carriers and nucleic acid lipid nanoparticle compositions (hereinafter referred to as "lipid nanoparticles").

[0223] 1. Animal Experiment Design

[0224] mRNA, sgRNA delivery experiment for base editor ABE8e targeting PCSK9

[0225] Cholesterol in the blood is primarily synthesized by the liver, which is also the primary organ responsible for breaking down excess cholesterol. On the surface of the liver, there is a low-density lipoprotein (LDL) receptor (LDLR). The LDL receptor binds to cholesterol circulating back to the liver, breaking it down into bile acids for excretion through the intestines. PCSK9 is a protease synthesized by the liver that binds to the LDL receptor, promoting its entry into liver cells, leading to its degradation by lysosomes and a decrease in its number. Therefore, inhibiting the activity of the enzyme PSCK9 can increase the number of LDLRs, thereby enhancing cholesterol uptake and breakdown. Basic and clinical studies have shown that the PCSK9 gene is an effective target for the treatment of hyperlipidemia and atherosclerosis. Figure 1 The figure shows the changes in the number of LDL receptors and the resulting changes in cholesterol metabolism before and after editing a specific site in the PCSK9 gene.

[0226] Strategies for PCSK9 gene editing and delivery in mouse liver cells Figure 1As shown, the main process is as follows: through intravenous injection, the prepared lipid nanoparticles are used to deliver the mRNA and sgRNA encoding ABE8e to mouse liver cells in a targeted manner. Under the action of ABE8e and sgRNA, mutations are introduced into the PCSK9 gene, and the mutation of base A to G is achieved at a specific site. The editing efficiency is calculated by sequencing.

[0227] The specific experimental design is as follows:

[0228] 1.1. Select appropriate mutation sites and edit design

[0229] The single-base editor ABE8e achieves precise A-to-G base substitutions without the need for a donor template and without causing DSBs. Based on this, the first exon of the PCSK9 gene was selected as a mutation site for screening. The mRNA encoding the single-base editor ABE8e and sgRNA were co-delivered into animals via lipid nanoparticles. The mRNA encoding the base editor ABE8e is translated into protein in the cytoplasm, forming a complex with the sgRNA before entering the cell nucleus. Under the guidance of the sgRNA, the base editor ABE8e targets the splice donor site in the first exon of the PCSK9 gene, deaminating the adenine (A) on the first exon template chain to inosine (I). I is read and replicated as G at the DNA level, ultimately achieving the A-to-G substitution, thereby destroying the splice donor site and prematurely terminating the PCSK9 gene reading frame.

[0230] 1.2. Preparation of mRNA and sgRNA for base editor ABE8e

[0231] The sequences of the first exon and the first intron of the mouse PCSK9 gene (NCBI Gene ID: 100102) were selected as the targeting region, and the target sequence PCSK9-sgRNA for single-base editing of the PCSK9 gene was determined.

[0232] By analyzing the sequence spanning the first exon and intron of the PCSK9 gene, an sgRNA targeting the region was designed: PCSK9-sgRNA (synthesized by Nanjing GenScript). The PCSK9-sgRNA sequence is:

[0233] PCSK9-sgRNA: 5'-CCCATACCTTGGAGCAACGG-3' (SEQ ID NO: 1);

[0234] sgRNAs were designed according to the target sequence and oligos were synthesized, the sgRNA sequence used is shown as SEQ ID NO: 1. CACC sequence was added at the 5' end of the upstream sequence of each sgRNA, and AAAC sequence was added at the 5' end of the downstream sequence. After synthesis, the upstream and downstream sequences were annealed by a preset program (95°C, 5 min; 95°C-85°C at -2°C / s; 85°C-25°C at -0.1°C / s; keep at 4°C), and the annealed product was ligated to the lenti U6-sgRNA / EF1a-mCherry vector (Addgene, Plasmid, #114199) linearized by BbsI (NEB, R3539S).

[0235] The system used in the construction of the sgRNA plasmid is as follows:

[0236] The linearization system of the lenti U6-sgRNA / EF1a-mCherry vector is as follows: 3 μg of vector; buffer (NEB: R0539L) 6 μL; BbsI 2 μL; ddH2O to 60 μL, 37°C enzyme digestion overnight.

[0237] The ligation system of the sgRNA annealing product and the linearized vector is as follows: T4 ligase buffer (NEB: M0202L) 1 μL, linearized vector 20 ng, annealed oligo fragment (10 μM) 5 μL, T4 ligase (NEB: M0202L) 0.5 μL, ddH2O to 10 μL, 16°C ligation overnight.

[0238] The ligated vector was transformed into E. coli DH5a competent cells (Weidi Biology, DL1001). The specific process is as follows: DH5a competent cells were taken out from -80°C and quickly inserted into ice, after 5 minutes, the bacterial block was melted, the ligation product was added and the bottom of the centrifuge tube was gently mixed by hand, and it was placed in ice for 25 minutes. 42°C water bath heat shock for 45 seconds, quickly put back into ice and stand for 2 minutes. 700 μL of sterile LB medium without antibiotics was added to the centrifuge tube, mixed and then incubated at 37°C, 200 rpm for 60 minutes. Centrifuge at 5000 rpm for 1 minute to collect the bacteria, take about 100 μL of supernatant, resuspend the bacterial block by gently blowing and spread on LB medium with Amp antibiotic. The plate was inverted and placed in a 37°C incubator for overnight culture. Single colonies were picked and confirmed by sequencing, then the positive clones were shaken and the plasmid was extracted (TIANGEN: DP120-01) to determine the concentration, and stored in a -20°C refrigerator for standby.

[0239] The base editor ABE8e used in this experiment is an efficient base editor ABE8e evolved by the David R. Liu team (Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR. "Phage-assisted evolution of anadenine base editor with improved Cas domain compatibility and activity", Nat Biotechnol., 2020, 38(7): 883-891. doi: 10.1038 / s41587-020-0453-z. Epub 2020Mar16. Erratumin: Nat Biotechnol. 2020May 20; PMID: 32433547; PMCID: PMC7357821). Plasmid ABE8e (Plasmid #138489) was purchased from Addgene, and ABE8e mRNA was expressed and purified in the laboratory for future use.

[0240] 2. Lipid nanoparticles were prepared by combining ionizable lipids (cationic lipids) or the compound of the present invention / DSPC / cholesterol / PEG-conjugated lipid PEG-DMG at a molar ratio of 50:10:38.5:1.5.

[0241] 2.1. Dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA, usually abbreviated as MC3), Compounds 1 to 5 of the present invention were dissolved in anhydrous ethanol with DSPC, cholesterol, and PEG-DMG according to the above molar ratios to obtain different lipid carriers.

[0242] 2.2. Ethanol solutions of different lipid carriers were mixed with mRNA buffer at a ratio of 1:3 (vol / vol) (the total lipid to mRNA mass ratio was 40:1, and the sgRNA:ABE8e mRNA (w / w) ratio was 1:1). Nucleic acid lipid nanoparticles 1-6 were prepared using a microfluidic nanomedicine manufacturing system (NanoAssemblr Ignite, Canada) at a flow rate of 12 mL / min. The obtained nucleic acid lipid nanoparticles were immediately diluted 40 times the volume into 1× DPBS buffer. The diluted nucleic acid lipid nanoparticle solution was concentrated to the desired volume by ultracentrifugation. After dilution, the solution was used for DLS particle size measurement and encapsulation efficiency testing.

[0243] 2.3. The particle size and polydispersity index (PDI) of the lipid nanoparticles were measured by dynamic light scattering using a Malvern Zetasizer Nano ZS (Malvern, UK) in 173° backscatter detection mode. The encapsulation efficiency of the lipid nanoparticles was determined using the Quant-it Ribogreen RNA Quantification Kit (ThermoFisher Scientific, UK) according to the manufacturer's instructions. The test results are shown in Table 1.

[0244] Table 1 Characterization of lipid nanoparticles

[0245] Lipid nanoparticles Cationic lipids Particle size (nm) PDI Encapsulation efficiency (%) 1 MC3 59.6 0.05 96.5 2 Compound 1 73.4 0.07 98.3 3 Compound 2 63.7 0.06 98.3 4 Compound 3 68.9 0.08 96.9 5 Compound 4 92.6 0.08 98.6 6 Compound 5 73.6 0.06 97.5

[0246] 3. In vivo editing experimental evaluation

[0247] 3.1. Lipid nanoparticles containing the compound of the present invention (see Table 1, lipid nanoparticles 2-6) encapsulating the mRNA and sgRNA encoding the base editor ABE8e were systemically administered to 6-7 week old C57BL / 6 female mice (purchased from Jiangsu Jicui Pharmaceutical Kang Co., Ltd.) at a dose of 0.2 mg / kg via tail vein injection. Lipid nanoparticle 1 containing dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA, abbreviated as MC3) encapsulating the mRNA and sgRNA of the base editor ABE8e was similarly administered to mice of the same age and sex as a positive control. In addition, PBS buffer was also injected into the tail vein of mice of the same age and sex in a similar manner as a negative control.

[0248] 3.2 Editing efficiency detection

[0249] The editing efficiency was tested one week after the mice were given the drug. The liver tissue was taken after the mice were killed, and the genome was extracted after lysis, and the efficiency was analyzed by deep sequencing.

[0250] The deep sequencing steps are as follows:

[0251] (1) Design primers according to the target gene location, see Table 2 for details.

[0252] Table 2 Design of primers targeting PCSK9 gene

[0253] Sequence objects Sequences PCSK9-F2 5'-ACCAGACGGCTAGATGAGCA-3' (SEQ ID NO: 2) PCSK9-R2 5'-CCCAGGACGAGGATGGAGATTA-3' (SEQ ID NO: 3)

[0254] (2) Editing efficiency detection.

[0255] The PCR program was as follows: 94°C for 2 minutes; 34 cycles of 98°C for 10 seconds, 60°C for 30 seconds, and 68°C for 20 seconds; and 68°C for 5 minutes. After PCR, gel electrophoresis was performed to confirm the amplification product by selecting a single band of appropriate size. The resulting PCR product was then sent to Nanjing GenScript for sequencing.

[0256] (3) The deep sequencing results were analyzed by Crispresso software for readout, specific site analysis, and editing efficiency calculation. The calculation results are shown in Table 3. The editing efficiency corresponding to each lipid nanoparticle can be seen in Table 3. Figure 2 .

[0257] Table 3 Evaluation of in vivo editing efficiency

[0258] Lipid nanoparticles Cationic lipids Editing efficiency (%) 1 MC3 20 2 Compound 1 35 3 Compound 2 18 4 Compound 3 26 5 Compound 4 10 6 Compound 5 11

[0259] As shown in Tables 1 and 3, the compounds (lipid compounds) provided by the present invention can effectively deliver drugs such as nucleic acid molecules and small molecule compounds; and by comparison, the lipid nanoparticles using the compounds of the present invention have a better particle size distribution, a high encapsulation efficiency, and a delivery effect that is significantly better than that of the comparative lipid nanoparticles, which can meet the needs of in vivo delivery.

[0260] The applicant states that while the present invention illustrates the lipid compounds and their applications through the aforementioned embodiments, the present invention is not limited to the aforementioned process steps, nor does it imply that the present invention must rely on the aforementioned process steps for implementation. Those skilled in the art will appreciate that any improvements to the present invention, equivalent substitutions for the raw materials used, additions of auxiliary components, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

Claims

1. A compound having the structure shown in Formula II-1 or a pharmaceutically acceptable form thereof, in, Dashed lines represent optional chemical bonds; R 1 、R 2 Each independently selected from C 1-6 Alkyl, or R 1 With R 2 Connected by chemical bonds to form a ring Cy, wherein the ring Cy is selected from a 3-6 membered heterocycle; G 1 Selected from C 2-6 alkylene; R 3 、R 5 、R 7 Each independently selected from C 2-16 alkylene; R 4 Selected from C 6-22 Straight-chain or branched alkyl group; R 6 selected from hydrogen; R 8 Selected from C 6-22 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The R 1 、R 2 Each independently selected from C 1-4 Alkyl, or R 1 With R 2 Connected by chemical bonds The wavy line represents the group and G 1 connection site.

3. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The G 1 Selected from The wavy line represents the attachment site of the group.

4. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The R 3 、R 5 、R 7 Each independently selected from The wavy line represents the attachment site of the group.

5. The compound according to claim 4 or a pharmaceutically acceptable form thereof, wherein The R 3 、R 7 Each independently selected from 6. The compound according to claim 4 or a pharmaceutically acceptable form thereof, wherein The R 5 Selected from 7. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The R 4 Selected from The wavy line represents the attachment site of the group.

8. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The R 8 Selected from The wavy line represents the attachment site of the group.

9. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The compound is selected from any one or a combination of at least two of Compound 1 to Compound 3: Compound 1 Compound 2 Compound 3 10. The compound according to claim 1 or a pharmaceutically acceptable form thereof, wherein The pharmaceutically acceptable form is selected from a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug.

11. Use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof in the preparation of liposome nanocarriers.

12. A lipid carrier comprising the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof.

13. The lipid carrier according to claim 12, wherein The lipid carrier comprises a combination of a first lipid compound and a second lipid compound; the first lipid compound comprises a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof and optionally other cationic lipids, and the second lipid compound comprises any one or a combination of at least two of anionic lipids, neutral lipids, steroids, and polymer-bound lipids.

14. A nucleic acid lipid nanoparticle composition comprising a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof, at least one of the lipid carriers according to claim 12 or 13, and a therapeutic agent or a prophylactic agent.

15. The nucleic acid lipid nanoparticle composition according to claim 14, wherein The therapeutic or prophylactic agent comprises a DNA, antisense nucleic acid, RNA, aptamer, ribozyme, immunostimulatory nucleic acid or PNA component.

16. The nucleic acid lipid nanoparticle composition according to claim 15, wherein The DNA includes a plasmid.

17. The nucleic acid lipid nanoparticle composition according to claim 15, wherein The antisense nucleic acid is an antisense oligonucleic acid.

18. The nucleic acid lipid nanoparticle composition according to claim 15, wherein The RNA includes mRNA, rRNA, circRNA, siRNA, saRNA, tRNA, snRNA, antagomir, microRNA inhibitor, microRNA activator or shRNA.

19. The nucleic acid lipid nanoparticle composition according to claim 15, wherein The RNA includes modified RNA.

20. The nucleic acid lipid nanoparticle composition according to claim 18, wherein The mRNA includes mRNA encoding an RNA-guided nuclease or an mRNA encoding a base editor, and a gRNA.

21. The nucleic acid lipid nanoparticle composition according to claim 20, wherein The nucleases include Cas9, Cas12, Cas13, IscB, TnpB, IsrB and homologs thereof.

22. A pharmaceutical formulation comprising at least one of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof, the lipid carrier according to claim 12 or 13, and the nucleic acid lipid nanoparticle composition according to any one of claims 14 to 21, and a pharmaceutically acceptable excipient.

23. The pharmaceutical preparation according to claim 22, wherein The pharmaceutically acceptable excipients include any one or a combination of at least two of excipients, carriers, diluents, fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, pH regulators, antioxidants, and buffers.

24. Use of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable form thereof, a lipid carrier according to claim 12 or 13, a nucleic acid lipid nanoparticle composition according to any one of claims 14 to 21, or a pharmaceutical preparation according to claim 22 or 23 in the preparation of a nucleic acid drug, a gene vaccine, a small molecule drug, a polypeptide or a protein drug.

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