Lipid compounds and lipid nanoparticles for delivery

Abstract

The invention discloses a compound with a structural formula shown in a formula (I), and salts and isomers thereof, and further discloses a nanoparticle composition containing the compound or the salts or the isomers thereof, and the nanoparticles can efficiently deliver active ingredients into cells to achieve the purpose of treatment or prevention. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to lipid compounds and lipid nanoparticle compositions for delivery. Background Art

[0002] The effective and targeted delivery of bioactive substances such as small molecule drugs, proteins and nucleic acids is an ongoing medical challenge. The key to the success of gene therapy is whether the therapeutic drugs can be safely and effectively delivered into the target cells through the vector in vivo. Due to the relative instability of nucleic acids and the low cell permeability of such substances, the delivery of nucleic acids to cells becomes difficult. Therefore, it is necessary to develop methods and compositions to promote the delivery of therapeutic and / or preventive drugs such as nucleic acids to cells. Gene therapy vectors are divided into viral vectors and non-viral vectors. Although viral vectors are used as efficient delivery systems to achieve target gene transfection and treatment purposes, viral vectors contain immunogenic viral proteins, limited target gene loading capacity and high prices. As a non-viral vector, lipid nanoparticles (LNPs) have received widespread attention due to their good in vitro stability, in vivo degradability, safety and reliability, and are widely used in gene therapy research for congenital and acquired genetic defects.

[0003] Lipid-containing nanoparticles or lipid nanoparticles, liposomes and lipid complexes have been shown to be effective transport carriers for bioactive substances such as small molecule drugs, proteins and nucleic acids entering and / or inside cells. LNPs refer to small vesicles formed by one or more lipid components, which can effectively compress and deliver various nucleic acid molecules, from DNA, RNA to chromosomes and even cells; LNPs are conducive to large-scale production due to their defined construction scheme and easy modification of targeting ligands.

[0004] LNPs generally include one or more cationic lipids and / or amino (ionizable) lipids, phospholipids containing polyunsaturated lipids, structural lipids (such as sterols) and / or lipids containing polyethylene glycol (PEG lipids). Cationic and / or ionizable lipids include, for example, lipids containing amines that can be easily protonated. Various such lipid-containing nanoparticle compositions have been shown in the prior art, such as patent document WO2017049245A2 discloses a compound and composition for intracellular delivery of therapeutic agents. Summary of the invention

[0005] Based on this, the present application discloses a lipid compound and a lipid nanoparticle composition comprising the compound, wherein the lipid has the advantages of high encapsulation rate, high expression, good targeting, etc.

[0006] Specifically, this application adopts the following technical solutions

[0007] 1. A compound of formula (I), or a salt thereof or an isomer thereof,

[0008]

[0009] Wherein R1 is vinyl, ethynyl or C1-C15 straight-chain alkyl, and R2, R3 and R4 are each independently C1-C15 straight-chain alkyl;

[0010] 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 C=O or O at the same time;

[0011] m, n, and o are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0012] 2. The compound according to item 1, or a salt thereof, or an isomer thereof, wherein R1 is a vinyl group or an ethynyl group.

[0013] 3. The compound according to item 1 or 2, or a salt thereof, or an isomer thereof, wherein R1 is a vinyl group.

[0014] 4. The compound according to any one of items 1 to 3, or a salt thereof, or an isomer thereof, wherein R2 is a C7-C11 straight-chain alkyl group. Preferably, R2 is a C9-C11 straight-chain alkyl group.

[0015] 5. The compound according to any one of items 1 to 4, or a salt thereof, or an isomer thereof, wherein R2 is a C8 or C10 straight-chain alkyl group.

[0016] 6. The compound according to any one of items 1 to 5, or a salt thereof, or an isomer thereof, wherein R3 and R4 are each independently a C6-C9 straight-chain alkyl group. Preferably, R3 and R4 are each independently a C7-C9 straight-chain alkyl group.

[0017] 7. The compound according to any one of items 1 to 6, or a salt thereof, or an isomer thereof, wherein R3 and R4 are each independently a C8 straight-chain alkyl group.

[0018] 8. The compound according to any one of items 1 to 7, or a salt thereof, or an isomer thereof, wherein m is selected from 3, 4, 5, 6, 7, and 8.

[0019] 9. The compound according to any one of items 1 to 8, or a salt thereof, or an isomer thereof, wherein m is selected from 4, 5, 6, 7, preferably, m is 5 or 7.

[0020] 10. The compound according to any one of items 1 to 9, or a salt thereof, or an isomer thereof, wherein n is selected from 3, 4, 5, 6, 7, 8.

[0021] 11. The compound according to any one of items 1 to 10, or a salt thereof, or an isomer thereof, wherein n is selected from 4, 5, 6, and 7, and preferably n is 7.

[0022] 12. The compound according to any one of items 1 to 11, or a salt thereof, or an isomer thereof, wherein o is selected from 2, 3, 4, 5, and 6.

[0023] 13. The compound according to any one of items 1 to 12, or its salt or isomer thereof, wherein X1 and X2 are both C=O, Y1 and Y2 are both O, or X1 is C=O, Y1 is O, X2 is O, and Y2 is C=0, or X1 is O, Y1 is C=O, X2 is C=O, and Y2 is O, or X1 is O, X2 is O, Y1 is C=O, and Y2 is C=O.

[0024] 14. A compound according to any one of items 1 to 13, or a salt thereof, or an isomer thereof, wherein m is selected from 5, 6, 7; n is selected from 5, 6, 7; o is selected from 2, 3, 4, 5, 6; R1 is vinyl; R2 is a C7-C11 straight-chain alkyl group (preferably R2 is a C9-C11 straight-chain alkyl group); R3 and R4 are each independently a C6-C9 straight-chain alkyl group (preferably R3 and R4 are each independently a C7-C9 straight-chain alkyl group); X1 and X2 are both C=O, Y1 and Y2 are both O or X1 is C=O, Y1 is O, X2 is O, and Y2 is C=0 or X1 is O, Y1 is C=O, X2 is C=O, and Y2 is O or X1 is O, X2 is O, Y1 is C=O, and Y2 is C=O.

[0025] Further preferably, m is selected from 5 or 7; n is selected from 7; o is selected from 2, 3, 4, 5, 6; R1 is vinyl; R2 is C8 or C10 straight chain alkyl; R3 and R4 are each independently C8 straight chain alkyl; X1 and X2 are both C=O, and Y1 and Y2 are both O.

[0026] 15. The compound according to item 1, or a salt thereof, or an isomer thereof, wherein the compound of formula (I) is selected from

[0027]

[0028]

[0029]

[0030]

[0031] 16. A nanoparticle composition comprising a lipid component, wherein the lipid component comprises the compound or salt thereof or isomer thereof according to any one of items 1 to 15.

[0032] 17. A nanoparticle composition according to claim 16, wherein the lipid component further comprises a phospholipid.

[0033] 18. The nanoparticle composition according to claim 17, wherein the phospholipid is selected from one or more of the following compounds:

[0034] Dilauroyl phosphatidylcholine (DLPC),

[0035] Dimyristoylphosphatidylcholine (DMPC),

[0036] Dioleoylphosphatidylcholine (DOPC),

[0037] Dipalmitoylphosphatidylcholine (DPPC),

[0038] Distearoylphosphatidylcholine (DSPC),

[0039] Dioleoylphosphatidylcholine (DUPC),

[0040] Palmitoyloleoylphosphatidylcholine (POPC),

[0041] 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0Diether PC),

[0042] 1-oleoyl-2-cholesteryl dimethylsuccinate-sn-glycero-3-phosphocholine (OChemsPC),

[0043] l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC),

[0044] 1,2-Divinyl-sn-glycero-3-phosphocholine,

[0045] 1,2-Diaryl acyl-sn-glycero-3-phosphocholine,

[0046] 1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE),

[0047] 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine,

[0048] 1,2-Divinyl alcohol-sn-glycero-3-phosphoethanolamine,

[0049] 1,2-Divinyl-sn-glycero-3-phosphoethanolamine,

[0050] 1,2-Diaryl-sn-glycero-3-phosphoethanolamine,

[0051] 1,2-dithiohexaenoic acid-sn-glycero-3-phosphoethanolamine,

[0052] 1,2-Dioleoyl-sn-glycero-3-phospho-(1-glycerol) sodium salt (DOPG) or sphingomyelin.

[0053] 19. A nanoparticle composition according to claim 17, wherein the phospholipid is DOPE.

[0054] 20. The nanoparticle composition of claim 17, wherein the phospholipid is DSPC.

[0055] 21. A nanoparticle composition according to any one of items 16-20, wherein the lipid component further comprises structural lipids.

[0056] 22. The nanoparticle composition of claim 21, wherein the structural lipid is selected from one or more of cholesterol, coprostanol, sitosterol, ergosterol, and stigmasterol.

[0057] 23. A nanoparticle composition according to any one of item 21, wherein the structural lipid is cholesterol.

[0058] 24. A nanoparticle composition according to any one of items 16-23, wherein the lipid component further comprises PEG lipids.

[0059] 25. A nanoparticle composition according to any one of claim 24, 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.

[0060] 26. A nanoparticle composition according to any one of items 16-25, wherein the lipid component further comprises cationic and / or ionizable lipids.

[0061] 27. The nanoparticle composition according to any one of items 16-26, further comprising a therapeutic and / or prophylactic agent selected from a vaccine or a compound capable of eliciting an immune response, a nucleic acid,

[0062] Preferably, the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

[0063] 28. A nanoparticle composition according to any one of items 16-27, wherein the encapsulation efficiency of the therapeutic agent and / or prophylactic agent is ≥50%; or ≥80%; or ≥90%.

[0064] 29. The nanoparticle composition of any one of items 16-28, wherein the average particle size of the nanoparticle composition is 50 nm-110 nm.

[0065] 30. The nanoparticle composition of any one of items 16-28, wherein the nanoparticle composition has a dispersibility index of 0.04-0.20.

[0066] 31. Use of a compound according to any one of items 1 to 15 in the preparation of a lipid nanoparticle composition.

[0067] 32. A pharmaceutical composition comprising the nanoparticle composition of any one of items 16-30 and a pharmaceutically acceptable carrier.

[0068] 33. A method for delivering a therapeutic and / or prophylactic agent to mammalian cells, the method comprising administering to a subject the nanoparticle composition of any one of items 16-30 or the pharmaceutical composition of item 32, wherein the administration comprises contacting the cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and / or prophylactic agent to the cells.

[0069] 34. A method according to claim 33, wherein the mammalian cell is in a mammal.

[0070] 35. The method of item 33 or 34, wherein the mammal is a human.

[0071] 36. A method according to any one of items 33-35, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0072] 37. A method for producing a target polypeptide in a mammalian cell, the method comprising contacting the cell with the nanoparticle composition described in any one of items 16-30 or the pharmaceutical composition described in item 32 to deliver a therapeutic and / or prophylactic agent to the cell, wherein the therapeutic and / or prophylactic agent is mRNA, which encodes the target polypeptide, such that the mRNA can be translated in the cell to produce the target polypeptide.

[0073] 38. The method of claim 37, wherein the mammalian cell is in a mammal.

[0074] 39. A method according to any one of items 37 or 38, wherein the mammalian cell is human.

[0075] 40. The method of any one of items 37-39, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0076] 41. A method for treating a disease or condition in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition of any one of items 16-30 or the pharmaceutical composition of item 31.

[0077] 42. A method according to claim 41, wherein the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity.

[0078] 43. A method according to item 41 or 42, wherein the disease or condition is selected from an infectious disease, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases or metabolic diseases.

[0079] 44. A method according to any one of items 41-43, wherein the mammal is a human.

[0080] 45. The method of any one of items 41-44, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0081] 46. ​​A method for specifically delivering a therapeutic and / or prophylactic agent to a mammalian organ, the method comprising administering to the mammal the nanoparticle composition described in any one of items 16-30 or the pharmaceutical composition described in item 32, wherein the administration comprises contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the organ.

[0082] 47. The method of claim 46, wherein the mammal is a human.

[0083] 48. The method of item 46 or 47, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0084] 49. A method according to any one of items 46-48, wherein the mammal is pretreated 24 hours or less prior to the contacting or administering step.

[0085] 50. The method of any one of items 46-49, wherein the mammal is pretreated about one hour prior to the contacting or administering step.

[0086] Technical Effects

[0087] The compounds of the present application can be used for the preparation of lipid nanoparticles. The nanoparticle compositions containing the compounds provided in the present application can achieve the encapsulation and delivery of therapeutic / prophylactic agents, safely deliver the therapeutic / prophylactic agents to the targeted locations, achieve high expression, and exert the effects of the therapeutic / prophylactic agents.

[0088] The lipid nanoparticles prepared in the present application have a small average particle size, a high encapsulation rate, and good targeting, and have broad application prospects in the field of drug delivery. BRIEF DESCRIPTION OF THE DRAWINGS

[0089] In order to more clearly illustrate the specific implementation of the present invention or the technical solution in the prior art, the drawings required for use in the specific implementation or the description of the prior art are briefly introduced below.

[0090] Figure 1 Shown are the normalized results of luciferase fluorescence intensity displayed by different LNP formulations 6 hours after intravenous injection.

[0091] Figure 2 Shown are the luciferase fluorescence intensities of different LNP formulations at 6 hours after intramuscular injection. DETAILED DESCRIPTION

[0092] The following is a description of the exemplary embodiments of the present application, including various details of the embodiments of the present application to facilitate understanding, which should be considered as merely exemplary. Therefore, it should be recognized by those of ordinary skill in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for clarity and conciseness, the description of well-known functions and structures is omitted in the following description.

[0093] Terms and Definitions

[0094] The present application discloses a compound of formula (I), or a salt thereof or an isomer thereof (preferably a stereoisomer),

[0095]

[0096] Wherein R1 is vinyl, ethynyl or C1-C15 straight-chain alkyl, and R2, R3 and R4 are each independently C1-C15 straight-chain alkyl;

[0097] 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 C=O or O at the same time;

[0098] m, n, and o are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0099] In a preferred embodiment, in the compound of formula (I), R1 is vinyl or ethynyl. In a further preferred embodiment, R1 is vinyl.

[0100] In a preferred embodiment, R2 is selected from C1-C15 straight chain alkyl, for example, R2 is selected from C2-C11 straight chain alkyl, R2 is selected from C3-C11 straight chain alkyl, R2 is selected from C4-C11 straight chain alkyl, R2 is selected from C5-C11 straight chain alkyl, R2 is selected from C6-C11 straight chain alkyl, R2 is selected from C7-C11 straight chain alkyl, R2 is selected from C8-C11 straight chain alkyl, R2 is selected from C9-C11 straight chain alkyl, R2 is selected from C10-C11 straight chain alkyl Alkyl, for example, R2 is C1 straight chain alkyl, R2 is C2 straight chain alkyl, R2 is C3 straight chain alkyl, R2 is C4 straight chain alkyl, R2 is C5 straight chain alkyl, R2 is C6 straight chain alkyl, R2 is C7 straight chain alkyl, R2 is C8 straight chain alkyl, R2 is C9 straight chain alkyl, R2 is C10 straight chain alkyl, R2 is C11 straight chain alkyl, R2 is C12 straight chain alkyl, R2 is C13 straight chain alkyl, R2 is C14 straight chain alkyl, R2 is C15 straight chain alkyl. In a further preferred embodiment, R2 is C10 straight chain alkyl. In a further preferred embodiment, R2 is C8 straight chain alkyl.

[0101] In a preferred embodiment, R3 is selected from C1-C15 straight chain alkyl, for example, R3 is selected from C2-C11 straight chain alkyl, R3 is selected from C3-C11 straight chain alkyl, R3 is selected from C4-C11 straight chain alkyl, R3 is selected from C5-C11 straight chain alkyl, R3 is selected from C6-C11 straight chain alkyl, R3 is selected from C7-C11 straight chain alkyl, R3 is selected from C8-C11 straight chain alkyl, R3 is selected from C9-C11 straight chain alkyl, R3 is selected from C10-C11 straight chain alkyl Alkyl, for example, R3 is C1 straight chain alkyl, R3 is C2 straight chain alkyl, R3 is C3 straight chain alkyl, R3 is C4 straight chain alkyl, R3 is C5 straight chain alkyl, R3 is C6 straight chain alkyl, R3 is C7 straight chain alkyl, R3 is C8 straight chain alkyl, R3 is C9 straight chain alkyl, R3 is C10 straight chain alkyl, R3 is C11 straight chain alkyl, R3 is C12 straight chain alkyl, R3 is C13 straight chain alkyl, R3 is C14 straight chain alkyl, R3 is C15 straight chain alkyl. In a further preferred embodiment, R3 is C8 straight chain alkyl.

[0102] In a preferred embodiment, R4 is selected from C1-C15 straight chain alkyl, for example, R4 is selected from C2-C11 straight chain alkyl, R4 is selected from C3-C11 straight chain alkyl, R4 is selected from C4-C11 straight chain alkyl, R4 is selected from C5-C11 straight chain alkyl, R4 is selected from C6-C11 straight chain alkyl, R4 is selected from C7-C11 straight chain alkyl, R4 is selected from C8-C11 straight chain alkyl, R4 is selected from C9-C11 straight chain alkyl, R4 is selected from C10-C11 straight chain alkyl Alkyl, for example, R4 is C1 straight chain alkyl, R4 is C2 straight chain alkyl, R4 is C3 straight chain alkyl, R4 is C4 straight chain alkyl, R4 is C5 straight chain alkyl, R4 is C6 straight chain alkyl, R4 is C7 straight chain alkyl, R4 is C8 straight chain alkyl, R4 is C9 straight chain alkyl, R4 is C10 straight chain alkyl, R4 is C11 straight chain alkyl, R4 is C12 straight chain alkyl, R4 is C13 straight chain alkyl, R4 is C14 straight chain alkyl, R4 is C15 straight chain alkyl. In a further preferred embodiment, R4 is C8 straight chain alkyl.

[0103] In a preferred embodiment, in the compound of formula (I), m is 3, 4, 5, 6, 7, or 8. In a further preferred embodiment, m is 4, 5, 6, or 7. In a further preferred embodiment, m is 5 or 7.

[0104] In a preferred embodiment, in the compound of formula (I), n is 3, 4, 5, 6, 7, or 8. In a further preferred embodiment, n is 4, 5, 6, or 7. In a further preferred embodiment, n is 7.

[0105] In a preferred embodiment, in the compound of the above formula (I), o is 2, 3, 4, 5, or 6.

[0106] In a preferred embodiment, in the compound of the above formula (I), X1 and X2 are both C=O, Y1 and Y2 are both O, or X1 is C=O, Y1 is O, X2 is O, Y2 is C=0, or X1 is O, Y1 is C=O, X2 is C=O, Y2 is O, or X1 is O, X2 is O, Y1 is C=O, Y2 is C=O.

[0107] In a preferred embodiment, in the compound of the above formula (I), m is selected from 5, 6, 7; n is selected from 5, 6, 7; o is selected from 2, 3, 4, 5, 6; R1 is vinyl; R2 is a C7-C11 straight-chain alkyl group (preferably R2 is a C9-C11 straight-chain alkyl group); R3 and R4 are each independently a C6-C9 straight-chain alkyl group (preferably R3 and R4 are each independently a C7-C9 straight-chain alkyl group); X1 and X2 are both C=O, Y1 and Y2 are both O or X1 is C=O, Y1 is O, X2 is O, Y2 is C=0 or X1 is O, Y1 is C=O, X2 is C=O, Y2 is O or X1 is O, X2 is O, Y1 is C=O, Y2 is C=O.

[0108] Further preferably, m is selected from 5 or 7; n is selected from 7; o is selected from 2, 3, 4, 5, 6; R1 is vinyl; R2 is C8 or C10 straight chain alkyl; R3 and R4 are each independently C8 straight chain alkyl; X1 and X2 are both C=O, and Y1 and Y2 are both O.

[0109] In a preferred embodiment, the compound of formula (I) is compound 1

[0110]

[0111] In a preferred embodiment, the compound of formula (I) is compound 2

[0112]

[0113] In a preferred embodiment, the compound of formula (I) is compound 3

[0114]

[0115] In a preferred embodiment, the compound of formula (I) is compound 4

[0116]

[0117] In a preferred embodiment, the compound of formula (I) is compound 5

[0118]

[0119] In a preferred embodiment, the compound of formula (I) is compound 6

[0120]

[0121] In a preferred embodiment, the compound of formula (I) is compound 7

[0122]

[0123] In a preferred embodiment, the compound of formula (I) is compound 8

[0124]

[0125] In a preferred embodiment, the compound of formula (I) is compound 9

[0126]

[0127] In a preferred embodiment, the compound of formula (I) is compound 10

[0128]

[0129] In a preferred embodiment, the compound of formula (I) is compound 11

[0130]

[0131] In a preferred embodiment, the compound of formula (I) is compound 12

[0132]

[0133] In a preferred embodiment, the compound of formula (I) is compound 13

[0134]

[0135] In a preferred embodiment, the compound of formula (I) is compound 14

[0136]

[0137] In a preferred embodiment, the compound of formula (I) is compound 15

[0138]

[0139] In a preferred embodiment, the compound of formula (I) is compound 16

[0140]

[0141] In a preferred embodiment, the compound of formula (I) is compound 17

[0142]

[0143] In a preferred embodiment, the compound of formula (I) is compound 18

[0144]

[0145] In a preferred embodiment, the compound of formula (I) is compound 19

[0146]

[0147] In a preferred embodiment, the compound of formula (I) is compound 20

[0148]

[0149] In a preferred embodiment, the compound of formula (I) is compound 21

[0150]

[0151] In a preferred embodiment, the compound of formula (I) is compound 22

[0152]

[0153] In a preferred embodiment, the compound of formula (I) is compound 23

[0154]

[0155] In a preferred embodiment, the compound of formula (I) is compound 24

[0156]

[0157] In a preferred embodiment, the compound of formula (I) is compound 25

[0158]

[0159] In a preferred embodiment, the compound of formula (I) is compound 26

[0160]

[0161] In a preferred embodiment, the compound of formula (I) is compound 27

[0162]

[0163] The present application further provides a nanoparticle composition comprising a lipid component, wherein the lipid component comprises a compound of formula (I) or a salt thereof or an isomer thereof provided by the present application.

[0164] In some embodiments, the nanoparticle composition has an average particle size of 50 nm to 110 nm.

[0165] The nanoparticle compositions may include, for example, lipid nanoparticles (LNPs), liposomes, lipid vesicles, and lipid complexes.

[0166] The nanoparticle compositions described herein include a lipid component comprising at least one compound according to formula (I). For example, the lipid component of the nanoparticle composition may include one or more of compounds 1-27. The nanoparticle composition may also include a variety of other components. For example, in addition to the compound according to formula (I), the lipid component of the nanoparticle composition may also include one or more other lipids.

[0167] The lipid component of the nanoparticle composition may include one or more PEG or PEG-modified lipids. Such substances may alternatively be referred to as PEGylated 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.

[0168] The lipid component of the nanoparticle composition can include one or more structural lipids. The structural lipid can be selected from, but not limited to, cholesterol, coprostanol, sitosterol, ergosterol, stigmasterol and mixtures thereof, but not limited thereto. 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 a combination thereof.

[0169] The lipid component of the nanoparticle composition may include one or more phospholipids. The phospholipids used in the nanoparticle composition and method may be selected from

[0170] Dilauroyl phosphatidylcholine (DLPC),

[0171] Dimyristoylphosphatidylcholine (DMPC),

[0172] Dioleoylphosphatidylcholine (DOPC),

[0173] Dipalmitoylphosphatidylcholine (DPPC),

[0174] Distearoylphosphatidylcholine (DSPC),

[0175] Dioleoylphosphatidylcholine (DUPC),

[0176] Palmitoyloleoylphosphatidylcholine (POPC),

[0177] 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0Diether PC),

[0178] 1-oleoyl-2-cholesteryl dimethylsuccinate-sn-glycero-3-phosphocholine (OChemsPC),

[0179] l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC),

[0180] 1,2-Divinyl-sn-glycero-3-phosphocholine,

[0181] 1,2-Diaryl acyl-sn-glycero-3-phosphocholine,

[0182] 1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE),

[0183] 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine,

[0184] 1,2-Divinyl alcohol-sn-glycero-3-phosphoethanolamine,

[0185] 1,2-Divinyl-sn-glycero-3-phosphoethanolamine,

[0186] 1,2-Diaryl-sn-glycero-3-phosphoethanolamine,

[0187] 1,2-dithiohexaenoic acid-sn-glycero-3-phosphoethanolamine,

[0188] 1,2-Dioleoyl-sn-glycero-3-phospho-(1-glycerol) sodium salt (DOPG) or sphingomyelin.

[0189] In some embodiments, the nanoparticle composition comprises DSPC.In some embodiments, the nanoparticle composition comprises DOPE.In some embodiments, the nanoparticle composition comprises DSPC and DOPE.

[0190] The nanoparticle composition may contain one or more therapeutic agents and / or preventive agents, and the therapeutic agents and / or preventive agents are 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.

[0191] The present application provides methods for delivering therapeutic and / or prophylactic agents to mammalian cells or organs, producing a polypeptide of interest in mammalian cells, and treating a disease or disorder in a mammal in need thereof, the method comprising administering to the mammal and / or contacting the mammalian cells with a therapeutic and / or prophylactic nanoparticle composition.

[0192] In certain embodiments, the therapeutic and / or preventive agent is mRNA. The mRNA can encode any polypeptide of interest, including any naturally or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA can have any size and can have any secondary structure or activity. In some embodiments, when expressed in a cell, the polypeptide encoded by the mRNA can have a therapeutic effect.

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

[0194] The nanoparticle composition can be characterized by a variety of methods. For example, a microscope (e.g., a transmission electron microscope or a scanning electron microscope) can be used to examine the morphology and size distribution of the nanoparticle composition. Dynamic light scattering or potentiometric methods (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-110 nm.

[0196] The nanoparticle composition can be relatively uniform. The polydispersity index can be used to indicate the uniformity of the nanoparticle composition, for example, the particle size distribution of the nanoparticle composition. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. The polydispersity index of the nanoparticle composition is 0.04-0.20.

[0197] The application further provides a method for delivering therapeutic and / or preventive agents specifically to mammalian organs, the method comprising administering any of the aforementioned nanoparticle compositions to a mammal, the administration comprising contacting a mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or preventive agent to the organ. Therapeutic and / or preventive agents, for example, proteins, cytotoxic agents, radioactive ions, chemotherapeutic agents or nucleic acids (e.g., RNA, e.g., mRNA) can be delivered to cells or organs. In the case of therapeutic and / or preventive being mRNA, when cells are contacted with the nanoparticle composition, translatable mRNA can be translated in cells to produce target polypeptides. However, substantially untranslatable mRNAs can also be delivered to cells. Substantially untranslatable mRNA can be used as a vaccine and / or can isolate the translation components of cells to reduce the expression of other species in the cell.

[0198] In some embodiments, the nanoparticle composition can target a specific type or category of cells (e.g., cells of a specific organ or system thereof). For example, a nanoparticle composition comprising a target treatment and / or prevention can be specifically delivered to the liver, kidney, spleen, femur or lung of a mammal. Specific delivery to a specific category of cells, organs or their systems or groups means that a higher proportion of the nanoparticle composition comprising a therapeutic and / or preventive agent, including treatment and / or prevention, is delivered to the target destination (e.g., tissue) relative to other destinations, for example, when administering a nanoparticle composition to a mammal. In some embodiments, the target tissue is selected from the liver, kidney, lung, spleen, femur, eye tissue (e.g., by intraocular, subretinal or intravitreal injection), vascular endothelium in a blood vessel (e.g., intracoronary or intrafemoral) or kidney, and tumor tissue (e.g., by intratumoral injection).

[0199] As used herein, the term "isomer" refers to any geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer or diastereomer of a compound. Compounds may contain one or more chiral centers and / or double bonds and may therefore exist as stereoisomers, such as double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis-trans isomers). The present application encompasses any and all isomers of the compounds described herein. Enantiomeric and stereoisomeric mixtures of compounds and methods of resolving them into their component enantiomers or stereoisomers are well known.

[0200] Example

[0201] Example 1: Synthesis of Compound 1

[0202]

[0203] The synthetic route is shown below.

[0204]

[0205] Specifically, compound B (5.07 g, 10.98 mmol) and DIEA (5.6 ml, 32.95 mmol) were added to a solution of compound A (12.87 g, 109.84 mmol) in EtOH (15 ml), stirred at room temperature for 3 hours, and the reaction was monitored by TLC. After the reaction was complete, the reaction solution was concentrated and subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to obtain compound C (5.03 g, 91.98%).

[0206] Under an ice-water bath and nitrogen atmosphere, add a solution of compound E (55 ml, 55 mmol) in ether to a solution of compound D (2.00 g, 45.40 mmol) in THF (80 ml) dropwise. After the addition is complete, stir at room temperature for 2 hours and monitor the reaction by TLC. After the reaction is complete, add a saturated ammonium chloride solution (80 mL) to the reaction solution, extract with dichloromethane (80 ml × 2), wash with saturated brine (80 ml), dry with anhydrous sodium sulfate, concentrate, and chromatograph on a silica gel column (petroleum ether: ethyl acetate = 30: 1) to obtain compound F (2.74 g, yield 32.4%).

[0207] Compound G (1.27 g, 5.37 mmol), DCC (1.33 g, 6.44 mmol) and DMAP (32.8 mg, 0.27 mmol) were added to a solution of compound F (1.10 g, 5.37 mmol) in dichloromethane (6 ml), and the mixture was stirred at room temperature for 3 hours. The reaction was monitored to be complete by TLC. The reaction solution was filtered, the filter cake was washed with DCM (5 ml), the filtrate was concentrated, and the compound H (1.88 g, yield: 87.9%) was obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 100: 1).

[0208] Compound C (300.0 mg, 0.60 mmol), K2CO3 (249.8 mg, 1.80 mmol) and KI (100.0 mg, 0.60 mmol) were added to a solution of compound H (219.0 mg, 0.60 mmol) in CPME / CH3CN (6 ml, 1V / 1V), and the mixture was heated to 70°C for 20 hours. The reaction was complete after TLC monitoring. The reaction solution was filtered, the filtrate was concentrated, and the mixture was subjected to silica gel column chromatography (MeOH: DCM = 20:1) to obtain compound 1 (260.0 mg, yield: 55.3%). 1H NMR(400MHz, CDCl3) δ4.95–4.80(m,2H),3.69(t,J=6.4Hz,2H),2.36–2.29(m,6H),1.7 2–1.46(m,21H),1.45–1.22(m,58H),0.92(t,J=8.0Hz,9H); MS-ESI(m / z):781.0(M+H) + .

[0209] Example 2: Synthesis of Compound 2

[0210]

[0211] The preparation method is the same as compound 1, and compound 2 can be prepared by using propionaldehyde instead of compound D as the raw material. 1 H NMR (400MHz, CDCl3) δ4.95–4.80(m,2H),3.68(t,J=6.4Hz,2H),2.63–2.49(m,6H),2.32(q,J=7.8H z,4H),1.73–1.47(m,20H),1.43–1.27(m,52H),0.91(t,J=8.0Hz,12H); MS-ESI(m / z):795.0(M+H) + .

[0212] Example 3: Synthesis of Compound 3

[0213]

[0214] The preparation method is the same as compound 1, and compound 3 can be prepared by using n-butyraldehyde instead of compound D as the raw material. 1 HNMR(400MHz, CDCl3) δ4.95–4.80(m,2H),3.70(t,J=6.4Hz,2H),2.34(t,J=7.8Hz,4H),1.69–1.58 (m,6H),1.58–1.50(m,10H),1.50–1.22(m,62H),0.94(t,J=8.0Hz,12H);MS-ESI(m / z):809.1(M+H) + .

[0215] Example 4: Synthesis of Compound 4

[0216]

[0217] The preparation method is the same as compound 1, and compound 4 can be prepared by using n-valeraldehyde instead of compound D as the raw material. 1HNMR(400MHz, CDCl3) δ4.95–4.80(m,2H),3.69(t,J=6.4Hz,2H),2.35(t,J=7.8Hz,4H),1.69–1.58 (m,6H),1.58–1.50(m,10H),1.50–1.22(m,64H),0.92(t,J=8.0Hz,12H);MS-ESI(m / z):823.0(M+H) + .

[0218] Example 5: Synthesis of Compound 5

[0219]

[0220] The preparation method is the same as compound 1, and compound 5 can be prepared by using acrolein instead of compound D as the raw material. 1 H NMR(400MHz, CDCl3)δ5.81(dd,J=13.6,8.0Hz,1H),5.32–5.15(m,3H),4.95–4.80(m,1H),3.69(t,J=6.4Hz,2H),2.63–2.49(m,6 H),2.34(t,J=7.8Hz,4H),1.74–1.59(m,10H),1.54(s,6H),1.45–1.27(m,54H),0.92(t,J=8.0Hz,9H); MS-ESI(m / z):793.1(M+H) + .

[0221] Example 6: Synthesis of Compound 6

[0222]

[0223] The synthetic route is shown below.

[0224]

[0225] Specifically, compound I (2.02 g, 7.80 mmol), DCC (1.93 g, 9.35 mmol) and DMAP (190.54 mg, 1.56 mmol) were added to a dichloromethane (8 ml) solution of compound G (1.52 g, 7.80 mmol), and the mixture was stirred at room temperature for 3 hours. The reaction was monitored to be complete by TLC. The reaction solution was filtered, the filter cake was washed with DCM (20 ml), the filtrate was concentrated, and the compound J (2.07 g, yield: 61.23%) was obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 100: 1).

[0226] Compound C (300.0 mg, 0.60 mmol), K2CO3 (249.8 mg, 1.80 mmol) and KI (100.0 mg, 0.60 mmol) were added to a solution of compound J (248.8 mg, 0.66 mmol) (248.8 mg, 0.66 mmol) in CPME / CH3CN (6 ml, 1V / 1V), and the mixture was heated to 70°C for 20 hours. The reaction was complete after TLC monitoring. The reaction solution was filtered, the filtrate was concentrated, and the compound 6 (0.39 g, yield: 76.10%) was obtained by silica gel column chromatography (MeOH: DCM = 20:1). 1 H NMR (400MHz, CDCl3) δ4.95–4.80(m,2H),3.72(t,J=6.8Hz,2H),2.63–2.49(m,6H),2.36(t,J=7.8 Hz,4H),1.73–1.59(m,7H),1.59–1.30(m,73H),0.92(t,J=8.0Hz,12H); MS-ESI(m / z):851.0(M+H) + .

[0227] Example 7: Synthesis of Compound 7

[0228]

[0229] The preparation method is the same as compound 6, and compound 7 can be prepared by using 8-bromooctanoic acid instead of compound G as the raw material. 1 H NMR (400MHz, CDCl3) δ4.95–4.80(m,2H),3.70(t,J=6.8Hz,2H),3.08-3.03(m,6H),2.32(t,J=8.0Hz,4H),1. 94–1.86(m,6H),1.67–1.48(m,16H),1.41–1.30(m,62H),0.92(t,J=8.0Hz,12H).MS-ESI(m / z):879.2(M+H) + .

[0230] Example 8: Synthesis of Compound 8

[0231]

[0232] The preparation method is the same as compound 1, and compound 8 can be prepared by using ethanolamine and acrolein instead of compound A and compound D as raw materials. 1H NMR (400MHz, CDCl3) δ5.87–5.74(m,1H),5.32–5.24(m,3H),4.99–4.82(m,1H),4.07–3.99(m,1H),3.22 –3.02(m,4H),2.47–2.24(m,4H),2.03–1.18(m,65H),0.92(t,J=8.0Hz,9H).MS-ESI(m / z):736.8(M+H) + .

[0233] Example 9: Synthesis of Compound 9

[0234]

[0235] The preparation method is the same as compound 1, and compound 9 can be prepared by using ethanolamine, acrolein and 8-bromooctanoic acid instead of compound A, compound D and compound G as raw materials. 1 H NMR (400MHz, CDCl3) δ6.00–5.64(m,1H),5.43–5.06(m,4H),4.22–4.11(m,1H),3.44(t,J=8.0Hz,4H),2.36 (t,J=7.6Hz,4H),1.94–1.85(m,4H),1.74–1.21(m,64H),0.92(t,J=6.8Hz,6H).MS-ESI(m / z):736.8(M+H) + .

[0236] Example 10: Synthesis of Compound 10

[0237]

[0238] The preparation method is the same as compound 1, using 3-amino-1-propanol and acrolein instead of compound A and compound D as raw materials to obtain compound 10. 1 H NMR (400MHz, CDCl3) δ5.88–5.74(m,1H),5.31–5.14(m,3H),4.97–4.84(m,1H),3.91(t,J=5.6Hz,2H),3. 37–2.96(m,6H),2.47–2.26(m,4H),2.15–1.19(m,64H),0.92(t,J=6.4Hz,9H).MS-ESI(m / z):750.8(M+H) + .

[0239] Example 11: Synthesis of Compound 11

[0240]

[0241] The preparation method is the same as compound 1, using 4-amino-1-butanol and acrolein instead of compound A and compound D as raw materials to obtain compound 11. 1 H NMR (400MHz, CDCl3) δ5.88–5.72(m,1H),5.38–5.12(m,3H),4.95–4.83(m,1H),3.81–3.70(m,1H),3.25 –2.85(m,4H),2.47–2.23(m,4H),2.07–1.15(m,69H),0.92(t,J=6.4Hz,9H).MS-ESI(m / z):764.8(M+H) + .

[0242] Example 12: Synthesis of Compound 15

[0243]

[0244] The synthetic route is as follows:

[0245]

[0246] Specifically, under an ice-water bath and nitrogen atmosphere, a solution of compound E (107 ml, 107 mmol) in ether was added dropwise to a solution of compound K (5.00 g, 89.19 mmol) in THF (50 ml), and after the addition was complete, the mixture was stirred at room temperature for 2 hours and monitored by TLC. After the reaction was complete, a saturated ammonium chloride solution (80 ml) was added to the reaction solution, and the mixture was extracted with dichloromethane (80 ml × 2), washed with saturated brine (80 ml), dried over anhydrous sodium sulfate, concentrated, and chromatographed on a silica gel column (petroleum ether: ethyl acetate = 20: 1) to obtain compound L (6.10 g, yield 30.53%).

[0247] Compound G (3.25 g, 16.64 mmol), DCC (3.74 g, 18.15 mmol) and DMAP (0.09 g, 0.76 mmol) were added to a solution of compound L (3.00 g, 15.13 mmol) in dichloromethane (30 ml), and the mixture was stirred at room temperature for 3 hours. The reaction was complete after monitoring by TLC. The reaction solution was filtered, the filter cake was washed with DCM (20 ml), the filtrate was concentrated, and the compound M (5.00 g, yield: 88.03%) was obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1).

[0248] Compound N (921.8 mg, 2.46 mmol) was added to a solution of compound M (100.0 mg, 1.64 mmol) in EtOH (10 ml), and the mixture was heated to 70°C for 16 hours and monitored by TLC. After the reaction was complete, the reaction solution was concentrated and subjected to silica gel column chromatography (dichloromethane: methanol = 25: 1) to give compound 15 (70.0 mg, 6.60%). 1 H NMR (400MHz, CDCl3) δ5.91–5.72(m,2H),5.30–5.16(m,6H),4.11–3.96(m,2H),3.26–3.01(m,6 H), 2.39 (t, J=7.2Hz, 4H), 2.07–1.18 (m, 48H), 0.92 (t, J=6.4Hz, 6H); MS-ESI (m / z): 650.7 (M+H) + .

[0249] Example 13: Synthesis of Compound 16

[0250]

[0251] The preparation method is the same as compound 1, using ethanolamine, compound M, acrolein, octyl magnesium bromide, 8-bromooctanoic acid instead of compound A, compound B, compound D, compound E and compound G as raw materials to obtain compound 16. 1 H NMR (400MHz, CDCl3) δ5.83–5.70(m,2H),5.26–5.11(m,6H),3.68(t,J=5.2Hz,2H),2.76(t,J=5.1Hz,2H),2.63(q,J=5.2Hz,3H),2 .31(q,J=7.2Hz,4H),1.60(ddd,J=27.1,14.6,7.3Hz,12H),1.45–1.12(m,38H),0.88(t,J=6.6Hz,6H).MS-ESI(m / z):650.5(M+H) + .

[0252] Example 14: Synthesis of Compound 17

[0253]

[0254] The preparation method is the same as compound 1, using propanolamine, acrolein and octyl magnesium bromide instead of compound A, compound D and compound E as raw materials to obtain compound 17. 1H NMR (400MHz, CDCl3) δ5.81(ddd,J=17.0,10.5,6.3Hz,1H),5.30–5.15(m,3H),4.90(p,J=6.3Hz,1H),3.84(t,J=5.2Hz,2H),2.80(d,J=6.0Hz,2H),2.59 (s,3H),2.34(dt,J=18.9,7.4Hz,4H),1.79(p,J=5.4Hz,2H),1.73–1.51(m, 14H),1.42–1.24(m,46H),0.92(t,J=6.8Hz,9H).MS-ESI(m / z):764.7(M+H) + .

[0255] Example 15: Synthesis of Compound 18

[0256]

[0257] The preparation method is the same as compound 1, using ethanolamine, acrolein and octylmagnesium bromide instead of compound A, compound D and compound E as raw materials to obtain compound 18. 1 H NMR(400MHz, CDCl3)δ5.77(ddd,J=17.0,10.5,6.3Hz,1H),5.26–5.12(m,3H), 4.86(p,J=6.3Hz,1H),3.53(s,1H),2.58(t,J=5.4Hz,2H),2.46(dq,J=8.3,4.9 Hz,4H),2.29(dt,J=15.3,7.5Hz,4H),1.62(dq,J=15.8,7.7Hz,6H),1.54–1.4 2(m,8H),1.38–1.18(m,46H),0.88(t,J=6.7Hz,9H).MS-ESI(m / z):764.7(M+H) + .

[0258] Example 16: Synthesis of Compound 19

[0259]

[0260] The preparation method is the same as compound 1, using ethanolamine, acrolein, octyl magnesium bromide and bromoacetic acid instead of compound A, compound D, compound E and compound G as raw materials to obtain compound 19. 1H NMR (400MHz, CDCl3) δ5.80 (ddd, J=17.2, 10.5, 6.7Hz, 1H), 5.35–5.16 (m, 3H), 4.90(p,J=6.2Hz,1H),3.58(t,J=5.2Hz,2H),3.40(s,2H),2.81(t,J=5.2Hz,2 H),2.70–2.61(m,2H),2.31(t,J=7.5Hz,2H),1.64(d,J=7.0Hz,2H),1.57–1.4 6(m,6H),1.37–1.26(m,44H),0.92(t,J=6.7Hz,9H).MS-ESI(m / z):652.6(M+H) + .

[0261] Example 17: Synthesis of Compound 20

[0262]

[0263] The preparation method is the same as compound 1, using ethanolamine, acrolein, octyl magnesium bromide and bromobutyric acid instead of compound A, compound D, compound E and compound G as raw materials to obtain compound 20. 1 H NMR (400MHz, CDCl3) δ5.80 (ddd, J=17.2, 10.5, 6.7Hz, 1H), 5.35–5.16 (m, 3H), 4.90(p,J=6.2Hz,1H),3.58(t,J=5.2Hz,2H),3.40(s,2H),2.81(t,J=5.2Hz,2 H),2.70–2.61(m,2H),2.31(t,J=7.5Hz,2H),1.64(d,J=7.0Hz,2H),1.57–1.4 6(m,6H),1.37–1.26(m,44H),0.92(t,J=6.7Hz,9H).MS-ESI(m / z):680.6(M+H) + .

[0264] Example 18: Synthesis of Compound 21

[0265]

[0266] The preparation method is the same as compound 15, using octylmagnesium bromide and 8-bromoheptanoic acid instead of compound E and compound G as raw materials to obtain compound 21. 1H NMR (400MHz, CDCl3) δ5.76(ddd,J=17.1,10.5,6.4Hz,2H),5.24–5.18(m,4H),5.15(dd,J=10.5,1.3Hz,2H),3.96(t,J=4.7Hz,2H),3.10(d,J=5.1Hz,2H) ,3.01(t,J=8.3Hz,4H),2.30(t,J=7.4Hz,4H),1.80(t,J=8.5Hz,4H),1.61( s,5H),1.50–1.07(m,40H),0.87(t,J=6.7Hz,6H).MS-ESI(m / z):650.5(M+H) + .

[0267] Example 19: Synthesis of Compound 23

[0268]

[0269] The preparation method is the same as compound 1, and compound 23 can be prepared by using ethanolamine, acrolein and bromobutyric acid instead of compound A, compound D and compound G as raw materials. 1 H NMR(400MHz, CDCl3)δ5.31–5.16(m,3H),2.59–2.49(m,4H),2.34(dt,J=24.5,7.4Hz,4H), 1.68–1.46(m,10H),1.36–1.26(m,46H),0.92(t,J=6.7Hz,9H).MS-ESI(m / z):708.7(M+H) + .

[0270] Example 20: Synthesis of Compound 24

[0271]

[0272] The preparation method is the same as compound 1, and compound 24 can be prepared by using acrolein, octylmagnesium bromide and bromoacetic acid instead of compound D, compound E and compound G as raw materials. 1 H NMR (400MHz, CDCl3) δ5.38–5.14(m,3H),3.68(t,J=6.6Hz,2H),3.36(s,2H),2.65–2.56(m,4H),2.31(t,J= 7.5Hz,2H),1.71–1.44(m,16H),1.32(d,J=15.5Hz,44H),0.92(t,J=6.7Hz,9H).MS-ESI(m / z):708.7(M+H) + .

[0273] Example 21: Synthesis of Compound 25

[0274]

[0275] The preparation method is the same as compound 1, using acrolein, octyl magnesium bromide and bromobutyric acid instead of compound D, compound E and compound G as raw materials to obtain compound 25. 1 H NMR (400MHz, CDCl3) δ5.31–5.15(m,3H),2.56–2.41(m,6H),2.35(dt,J=27.7,7.4Hz,4H), 1.66–1.48(m,12H),1.42–1.25(m,48H),0.92(t,J=6.7Hz,9H).MS-ESI(m / z):736.7(M+H) + .

[0276] Example 22: Synthesis of Compound 26

[0277]

[0278] The preparation method is the same as compound 1, using acrolein and bromoacetic acid instead of compound D and compound G as raw materials to obtain compound 26. 1 H NMR (400MHz, CDCl3) δ5.81(ddd,J=17.1,10.5,6.6Hz,1H),5.36–5.15(m,3H),4.90(p,J=6.3Hz,1H),3.68(t,J=6.6Hz,2H),3.36(s, 2H),2.67–2.54(m,4H),2.31(t,J=7.5Hz,2H),1.69–1.46(m,15H),1.39–1.27(m,49H),0.97–0.86(m,9H).MS-ESI(m / z):736.7(M+H) + .

[0279] Example 23: Synthesis of Compound 27

[0280]

[0281] The preparation method is the same as compound 1, using acrolein and bromobutyric acid instead of compound D and compound G as raw materials to obtain compound 27. 1H NMR (400MHz, CDCl3) δ5.31–5.14(m,3H),3.67(t,J=6.6Hz,2H),2.51–2.39(m,6H),2.33(dt,J=24.3,7 .5Hz,4H),1.79(p,J=7.4Hz,2H),1.67–1.27(m,64H),0.91(t,J=6.7Hz,9H).MS-ESI(m / z):764.7(M+H) + .

[0282] Test example

[0283] Experimental Example 1: Lipid Nanoparticle (LNP) Encapsulation

[0284] Disperse the Luciferase mRNA stock solution in 20mM acetic acid solution (pH5.3) to a final concentration of 200μg / mL (aqueous phase). Mix into a lipid mixture (oil phase) according to the molar ratio of Example Compound: Cholesterol: DSPC: DMG-PEG2000 = 50:38.5:10:1.5. Control the flow rate of the aqueous phase and the oil phase to mix the mRNA with the lipid mixture through T mixing to obtain LNP-encapsulated mRNA. Dilute the encapsulated LNP with buffer, then concentrate it by ultrafiltration, and replace the diluent. Finally, concentrate the LNP to 100μg / mL of mRNA, and adjust the pH of the LNP to about 7-8. Finally, use Ribogreen kit and 10% Triton as a demulsifier to detect the total content and free content of mRNA in LNP, and calculate the encapsulation rate of LNP. The final LNP product was diluted with diluent, 1 ml was added to the particle size pool, and placed on the Malvern ZetaSizer instrument to detect the particle size of the LNP. The results are shown in Table 1.

[0285] Particle size, PDI and encapsulation efficiency are important quality attributes of lipid nanoparticles. As shown in the table below, the tested compounds all have good encapsulation efficiency, particle size suitable for mRNA delivery and narrow PDI.

[0286] Table 1: LNP characterization data of example compounds

[0287] serial number Particle size (nm) PDI Encapsulation rate (%) Compound 1 70 0.072119 97.8 Compound 2 73.73 0.06921 98.6 Compound 3 67.94 0.07084 98.3 Compound 4 69.12 0.08419 96.2 Compound 5 76.7 0.1258 97.7 Compound 6 66.07 0.1005 97.1 Compound 7 61.93 0.1183 98.2 Compound 8 71.91 0.1078 98.54 Compound 9 79.52 0.1492 95.96 Compound 10 73.04 0.0923 98.62 Compound 11 87.65 0.1521 96.68 Compound 16 114 0.07024 66.81 Compound 17 78.28 0.09819 95.64 Compound 18 77.56 0.14530 97.83 Compound 19 98 0.1269 61.6 Compound 20 104.2 0.07026 74.9 Compound 21 100.1 0.11240 66.10 Compound 23 74.54 0.09163 96.7 Compound 24 99.8 0.1314 61.8 Compound 25 80.24 0.06869 96.3 Compound 26 100.5 0.1306 63.7 Compound 27 83.07 0.04973 97.2

[0288] Test Example 2: Intravenous injection delivery effect test

[0289] 1. Encapsulate the mRNA expressing Luciferase into the LNP formulations of compound MC3, compound 1, compound 2, compound 4, compound 5, compound 6, compounds 8-11, compounds 16-18, and compound 21. The LNP formulation preparation method and the mRNA encapsulation method are as described in Experimental Example 1;

[0290] 2. The loaded LNP formulation was injected intravenously into Balb / c mice at a dose of 1 mg / kg, with 5 mice in each group.

[0291] 3. Detect the Luciferase fluorescence expression intensity of each mouse at 6 hours.

[0292] 4. Detection of fluorescence expression intensity: D-luciferin sodium salt (dose: 150 mg / kg) was injected intraperitoneally into each mouse 10 minutes before the test. The mice were then anesthetized with isoflurane and placed in an IVIS instrument, and bioluminescence was selected for detection.

[0293] 5. Determine the delivery effect of the compound based on the fluorescence intensity results.

[0294] The delivery and expression effects of the LNP of the compound of the present invention are as follows Figure 1 The data show that the expression of compound 1 and compound 2 groups was slightly higher than that of MC3 group, the expression of compound 4, compound 6, compound 16 and compound 21 groups was lower than that of MC3 group, while the expression of compound 5, compound 8-11 and compound 17-18 groups was significantly higher than that of MC3, and the expression was two to four times that of MC3 group.

[0295] Test Example 3: Intramuscular injection delivery effect test

[0296] 1. Encapsulate the mRNA expressing Luciferase into the LNP formulations of compound MC3, compound 8, compound 10, and compound 11. The LNP formulation preparation method and the mRNA encapsulation method are as described in Experimental Example 1;

[0297] 2. The loaded LNP formulation was injected intramuscularly into Balb / c mice, 50ul per mouse, the injection dose was 5μg / mouse, 5 mice per group.

[0298] 3. Detect the Luciferase fluorescence expression intensity of each mouse at 6 hours.

[0299] 4. Detection of fluorescence expression intensity: D-luciferin sodium salt (dose: 150 mg / kg) was injected intraperitoneally into each mouse 10 minutes before the test. The mice were then anesthetized with isoflurane and placed in the IVIS instrument. Bioluminescence was selected for detection. After the detection was completed, the fluorescence intensity of the muscle injection site was analyzed.

[0300] 5. Determine the delivery effect of the compound based on the fluorescence intensity results.

[0301] according to Figure 2The results show that compound 8, compound 10 and compound 11 can also achieve good expression effects through intramuscular injection, which are significantly better than the positive reference group MC3, among which the expression advantage of compound 8 group is the most obvious.

[0302] Although the embodiments of the present application are described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely illustrative and instructive, rather than restrictive. A person of ordinary skill in the art can make many forms under the guidance of this specification and without departing from the scope of protection of the claims of the present application, all of which belong to the protection of the present application.

Claims

1. A compound of formula (I), or a salt thereof or a stereoisomer thereof, Wherein R1 is vinyl, ethynyl or C1-C15 straight-chain alkyl, and R2, R3 and R4 are each independently C1-C15 straight-chain alkyl; 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 C=O or O at the same time; m, n, and o are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

2. The compound or salt thereof or stereoisomer thereof according to claim 1, wherein: R1 is vinyl or ethynyl.

3. The compound or salt thereof or stereoisomer thereof according to claim 1 or 2, wherein: R1 is vinyl.

4. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 3, wherein: R2 is a C7-C11 straight chain alkyl group.

5. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 4, wherein: R2 is a C8 or C10 straight chain alkyl group.

6. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 5, wherein: R3 and R4 are each independently a C6-C9 straight-chain alkyl group.

7. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 6, wherein: R3 and R4 are each independently a C8 straight chain alkyl group.

8. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 7, wherein: m is selected from 3, 4, 5, 6, 7, 8.

9. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 8, wherein: m is selected from 4, 5, 6, and 7. Preferably, m is 5 or 7.

10. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 9, wherein: n is selected from 3, 4, 5, 6, 7, 8.

11. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 10, wherein: n is selected from 4, 5, 6, and 7. Preferably, n is 7.

12. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 11, wherein: o is selected from 2, 3, 4, 5, 6.

13. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 12, wherein: X1 and X2 are both C=O, Y1 and Y2 are both O, or X1 is C=O, Y1 is O, X2 is O, Y2 is C=0, or X1 is O, Y1 is C=O, X2 is C=O, Y2 is O, or X1 is O, X2 is O, Y1 is C=O, Y2 is C=O.

14. The compound or salt thereof or stereoisomer thereof according to any one of claims 1 to 13, wherein: m is selected from 5, 6, 7; n is selected from 5, 6, 7; o is selected from 2, 3, 4, 5, 6; R1 is vinyl; R2 is C7-C11 straight-chain alkyl; R3 and R4 are each independently C6-C9 straight-chain alkyl; X1 and X2 are both C=O, Y1 and Y2 are both O or X1 is C=O, Y1 is O, X2 is O, Y2 is C=0 or X1 is O, Y1 is C=O, X2 is C=O, Y2 is O or X1 is O, X2 is O, Y1 is C=O, Y2 is C=O.

15. The compound or salt thereof or stereoisomer thereof according to claim 1, wherein: The compound of formula (I) is selected from Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 Compound 6 Compound 7 Compound 8 Compound 9 Compound 10 Compound 11 Compound 12 Compound 13 Compound 14 Compound 15 Compound 16 Compound 17 Compound 18 Compound 19 Compound 20 Compound 21 Compound 22 Compound 23 Compound 24 Compound 25 Compound 26 Compound 27 16. A nanoparticle composition comprising a lipid component, wherein the lipid component comprises the compound or salt thereof or a stereoisomer thereof according to any one of claims 1 to 15.

17. The nanoparticle composition of claim 16, wherein the lipid component further comprises a phospholipid.

18. The nanoparticle composition according to claim 17, wherein the phospholipid is selected from one or more of the following compounds: Dilauroyl phosphatidylcholine (DLPC), Dimyristoylphosphatidylcholine (DMPC), Dioleoylphosphatidylcholine (DOPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dioleoylphosphatidylcholine (DUPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesteryl dimethylsuccinate-sn-glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-Divinyl-sn-glycero-3-phosphocholine, 1,2-Diaryl acyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE), 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-Divinyl alcohol-sn-glycero-3-phosphoethanolamine, 1,2-Divinyl-sn-glycero-3-phosphoethanolamine, 1,2-Diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexaenoic acid-sn-glycero-3-phosphoethanolamine, 1,2-Dioleoyl-sn-glycero-3-phospho-(1-glycerol) sodium salt (DOPG) or sphingomyelin.

19. The nanoparticle composition of claim 17, wherein the phospholipid is DOPE.

20. The nanoparticle composition of claim 17, wherein the phospholipid is DSPC.

21. The nanoparticle composition of any one of claims 16-20, wherein the lipid component further comprises a structural lipid.

22. The nanoparticle composition of claim 21, wherein the structural lipid is selected from one or more of cholesterol, coprostanol, sitosterol, ergosterol, and stigmasterol.

23. The nanoparticle composition of any one of claims 21, wherein the structural lipid is cholesterol.

24. The nanoparticle composition of any one of claims 16-23, wherein the lipid component further comprises a PEG lipid.

25. The nanoparticle composition of any one of claim 24, 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.

26. The nanoparticle composition of any one of claims 16-25, wherein the lipid component further comprises a cationic and / or ionizable lipid.

27. The nanoparticle composition according to any one of claims 16 to 26, further comprising a therapeutic and / or prophylactic agent selected from a vaccine or a compound capable of eliciting an immune response, a nucleic acid, Preferably, the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

28. The nanoparticle composition of any one of claims 16-27, wherein the encapsulation efficiency of the therapeutic agent and / or prophylactic agent is ≥50%; or ≥80%; or ≥90%.

29. The nanoparticle composition according to any one of claims 16-28, wherein the average particle size of the nanoparticle composition is 50 nm-110 nm.

30. The nanoparticle composition of any one of claims 16-28, wherein the nanoparticle composition has a dispersibility index of 0.04-0.

20.

31. Use of the compound of any one of claims 1-15 in the preparation of a lipid nanoparticle composition.

32. A pharmaceutical composition comprising the nanoparticle composition of any one of claims 16-30 and a pharmaceutically acceptable carrier.

33. A method of delivering a therapeutic and / or prophylactic agent to a mammalian cell, the method comprising administering to a subject the nanoparticle composition of any one of claims 16-30 or the pharmaceutical composition of claim 31, wherein the administration comprises contacting a cell with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and / or prophylactic agent to the cell.

34. The method of claim 33, wherein the mammalian cell is in a mammal.

35. The method of claim 33 or 34, wherein the mammal is a human.

36. The method of any one of claims 33-35, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

37. A method for producing a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the nanoparticle composition of any one of claims 16-30 or the pharmaceutical composition of claim 32 to deliver a therapeutic and / or prophylactic agent to the cell, wherein the therapeutic and / or prophylactic agent is mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest.

38. The method of claim 37, wherein the mammalian cell is in a mammal.

39. The method of any one of claims 37 or 38, wherein the mammalian cell is human.

40. The method of any one of claims 37-39, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

41. A method of treating a disease or condition in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition of any one of claims 16-30 or the pharmaceutical composition of claim 31.

42. The method of claim 41, wherein the disease or disorder is characterized by dysfunctional or aberrant protein or polypeptide activity.

43. The method of claim 41 or 42, wherein the disease or condition is selected from an infectious disease, cancer and proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a renal vascular disease, or a metabolic disease.

44. The method of any one of claims 41-43, wherein the mammal is a human.

45. The method of any one of claims 41-44, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

46. ​​A method for specifically delivering a therapeutic and / or prophylactic agent to a mammalian organ, the method comprising administering to the mammal the nanoparticle composition of any one of claims 16-30 or the pharmaceutical composition of claim 32, wherein the administration comprises contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the organ.

47. The method of claim 46, wherein the mammal is a human.

48. The method of claim 46 or 47, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

49. The method of any one of claims 46-48, wherein the mammal is pretreated 24 hours or less prior to the contacting or administering step.

50. The method of any one of claims 46-49, wherein the mammal is pretreated about one hour prior to the contacting or administering step.

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