Lipid Compounds and Lipid Nanoparticles for Delivery

Abstract

The present application discloses a compound, the structural formula of which is shown in formula (I), as well as its salts and isomers. The present application also discloses a nanoparticle composition comprising the above-mentioned compound or its salt or its isomer. The nanoparticles of the present application can efficiently deliver the active ingredient into cells to achieve therapeutic or prophylactic purposes.

Description

Technical Field

[0001] This application relates to the field of biotechnology, and specifically to lipid compounds and lipid nanoparticle compositions for delivery. Background Art

[0002] The effective 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 lies in whether therapeutic drugs can be safely and effectively delivered into target cells through a 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 is difficult. Therefore, methods and compositions need to be developed to facilitate the delivery of therapeutic and / or prophylactic drugs such as nucleic acids to cells. Gene therapy vectors are divided into viral vectors and non-viral vectors. Although viral vectors are efficient delivery systems for achieving transfection of target genes and therapeutic purposes, due to problems such as immunogenic viral proteins, limited loading capacity of target genes, and high costs, lipid nanoparticles (LNPs), as non-viral vectors, have received extensive attention due to their good in vitro stability, biodegradability in vivo, safety and reliability, etc., 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 proven to be effective transport carriers for bioactive substances such as small molecule drugs, proteins, and nucleic acids to enter cells and / or intracellularly. 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 schemes 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, which can be easily protonated. Various such lipid-containing nanoparticle compositions have been demonstrated in the prior art. For example, patent document WO2017049245A2 discloses a compound and composition for intracellular delivery of a therapeutic agent. Summary of the Invention

[0005] Based on this, this application discloses a lipid compound and a lipid nanoparticle composition containing the compound, and the lipid has advantages such as high encapsulation rate, high expression, and good targeting.

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

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

[0008]

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

[0010] X1 and X2 are independently selected from C=O or O, and Y1, Y2 are independently selected from C=O or O, provided that X1 and Y1, X2 and Y2 are not simultaneously C=O or O;

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

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

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

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

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

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

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

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

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

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

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

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

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

[0024] 14. The compound or its salt or its isomer according to any one of items 1 - 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 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); both X1 and X2 are C=O, both Y1 and Y2 are O, or X1 is C=O, Y1 is O, X2 is O, Y2 is C=O, 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.

[0025] More 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 a C8 or C10 straight-chain alkyl group; R3 and R4 are each independently a C8 straight-chain alkyl group; both X1 and X2 are C=O, both Y1 and Y2 are O.

[0026] 15. The compound or its salt or its isomer according to item 1, 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 its salt or its isomer according to any one of items 1 - 15.

[0032] 17. The nanoparticle composition according to item 16, wherein the lipid component further comprises phospholipids.

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

[0034] Dilauroyl phosphatidylcholine (DLPC),

[0035] Dimyristoyl phosphatidylcholine (DMPC),

[0036] Dioleoyl phosphatidylcholine (DOPC),

[0037] Dipalmitoyl phosphatidylcholine (DPPC),

[0038] Distearoyl phosphatidylcholine (DSPC),

[0039] Dioleoyl phosphatidylcholine (DUPC),

[0040] Palmitoyl oleoyl phosphatidylcholine (POPC),

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

[0042] 1 - Oleoyl - 2 - cholesteryl dimethyl succinate - sn - glycero - 3 - phosphocholine (OChemsPC),

[0043] 1 - Hexadecyl - sn - glycero - 3 - phosphocholine (C16 Lyso PC),

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

[0045] 1,2 - Diarachidonoyl - sn - glycero - 3 - phosphocholine,

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

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

[0048] 1,2 - Divinylol - sn - glycero - 3 - phosphoethanolamine,

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

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

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

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

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

[0054] 20. The nanoparticle composition according to item 17, wherein the phospholipid is DSPC.

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

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

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

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

[0059] 25. The nanoparticle composition according to any one of item 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. The nanoparticle composition according to any one of items 16-25, wherein the lipid component further comprises a cationic and / or ionizable lipid.

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

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

[0064] 29. The nanoparticle composition according to 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 according to any one of items 16 - 28, wherein the dispersity index of the nanoparticle composition is 0.04 - 0.20.

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

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

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

[0069] 34. The method according to item 33, wherein the mammalian cells are in a mammal.

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

[0071] 36. The 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 of producing a polypeptide of interest in mammalian cells, the method comprising contacting the cells with the nanoparticle composition according to any one of items 16 - 30 or the pharmaceutical composition according to item 32 to deliver a therapeutic agent and / or a prophylactic agent to the cells, wherein the therapeutic agent and / or the prophylactic agent is mRNA, the mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cells to produce the polypeptide of interest.

[0073] 38. The method according to item 37, wherein the mammalian cells are in a mammal.

[0074] 39. The method according to any one of items 37 or 38, wherein the mammalian cells are human.

[0075] 40. The method according to 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 disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition according to any one of items 16 - 30 or the pharmaceutical composition according to item 31.

[0077] 42. The method according to item 41, wherein the disease or disorder is characterized by a dysfunctional or abnormal protein or polypeptide activity.

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

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

[0080] 45. The method according to 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 agent and / or a prophylactic agent to an organ of a mammal, the method comprising administering to the mammal the nanoparticle composition according to any one of items 16 - 30 or the pharmaceutical composition according to item 32, the administration comprising bringing the mammalian organ into contact with the nanoparticle composition, thereby delivering the therapeutic agent and / or the prophylactic agent to the organ.

[0082] 47. The method according to item 46, wherein the mammal is a human.

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

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

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

[0086] Technical effect

[0087] The compounds of the present application can be used for the preparation of lipid nanoparticles. The nanoparticle composition containing the compounds provided by the present application can achieve the encapsulation and delivery of a therapeutic agent / prophylactic agent, safely deliver the therapeutic agent / prophylactic agent to the target position, achieve high expression, and exert the effect of the therapeutic agent / prophylactic agent.

[0088] The lipid nanoparticles prepared in this application have a small average particle size, a high encapsulation rate, and good targeting properties, 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 embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art.

[0090] Figure 1 Shows the normalized results of Luciferase fluorescence intensity shown by different LNP formulations 6 hours after intravenous injection.

[0091] Figure 2 Shows the Luciferase fluorescence intensity shown by different LNP formulations 6 hours after intramuscular injection. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0092] The following describes exemplary embodiments of the present application, including various details of the embodiments of the present application to facilitate understanding. It should be considered merely exemplary. Therefore, those of ordinary skill in the art should recognize 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 below omits the description of well-known functions and structures.

[0093] TERMS AND DEFINITIONS

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

[0095]

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

[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 simultaneously C=O or O;

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

[0099] In a preferred embodiment, in the compound of formula (I) above, 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 groups. For example, R2 is selected from C2-C11 straight-chain alkyl groups, R2 is selected from C3-C11 straight-chain alkyl groups, R2 is selected from C4-C11 straight-chain alkyl groups, R2 is selected from C5-C11 straight-chain alkyl groups, R2 is selected from C6-C11 straight-chain alkyl groups, R2 is selected from C7-C11 straight-chain alkyl groups, R2 is selected from C8-C11 straight-chain alkyl groups, R2 is selected from C9-C11 straight-chain alkyl groups, R2 is selected from C10-C11 straight-chain alkyl groups. For example, R2 is C1 straight-chain alkyl group, R2 is C2 straight-chain alkyl group, R2 is C3 straight-chain alkyl group, R2 is C4 straight-chain alkyl group, R2 is C5 straight-chain alkyl group, R2 is C6 straight-chain alkyl group, R2 is C7 straight-chain alkyl group, R2 is C8 straight-chain alkyl group, R2 is C9 straight-chain alkyl group, R2 is C10 straight-chain alkyl group, R2 is C11 straight-chain alkyl group, R2 is C12 straight-chain alkyl group, R2 is C13 straight-chain alkyl group, R2 is C14 straight-chain alkyl group, R2 is C15 straight-chain alkyl group. In a further preferred embodiment, R2 is C10 straight-chain alkyl group. In a further preferred embodiment, R2 is C8 straight-chain alkyl group.

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

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

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

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

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

[0106] In a preferred embodiment, in the compound of formula (I) above, both X1 and X2 are C=O, both Y1 and Y2 are 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=O 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] More 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 a C8 or C10 straight-chain alkyl group; R3 and R4 are each independently a C8 straight-chain alkyl group; X1 and X2 are both C=O, Y1 and Y2 are both O.

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

[0110]

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

[0112]

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

[0114]

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

[0116]

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

[0118]

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

[0120]

[0121] In a preferred embodiment, the compound of the 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 the compound of formula (I) provided by the present application, or a salt or an isomer thereof.

[0164] In some embodiments, the average particle size of the nanoparticle composition is 50 nm - 110 nm.

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

[0166] The nanoparticle composition described in this application contains a lipid component, and the lipid component contains 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 contain a variety of other components. For example, in addition to the compound according to formula (I), the lipid component of the nanoparticle composition may further include one or more other lipids.

[0167] The lipid component of the nanoparticle composition may include one or more PEGs or PEG-modified lipids. Such substances may alternatively be referred to as polyethylene glycolated 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, the PEG lipid may be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or PEG-DSPE lipid.

[0168] The lipid component of the nanoparticle composition may include one or more structural lipids. The structural lipids may be selected from, but are not limited to, cholesterol, coprostanol, sitosterol, ergosterol, stigmasterol, and mixtures thereof, but are not limited thereto. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid includes cholesterol and corticosteroids (such as prednisolone, dexamethasone, prednisone, and hydrocortisone), or combinations thereof.

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

[0170] dilauroyl phosphatidylcholine (DLPC),

[0171] dimyristoyl phosphatidylcholine (DMPC),

[0172] dioleoyl phosphatidylcholine (DOPC),

[0173] dipalmitoyl phosphatidylcholine (DPPC),

[0174] distearoyl phosphatidylcholine (DSPC),

[0175] dioleoyl phosphatidylcholine (DUPC),

[0176] Palmitoyl oleoyl phosphatidylcholine (POPC),

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

[0178] 1 - Oleoyl - 2 - cholesteryl dimethyl succinate - sn - glycero - 3 - phosphocholine (OChemsPC),

[0179] 1 - Hexadecyl - sn - glycero - 3 - phosphocholine (C16 Lyso PC),

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

[0181] 1,2 - Diarachidonoyl - sn - glycero - 3 - phosphocholine,

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

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

[0184] 1,2 - Divinylacetyl - sn - glycero - 3 - phosphoethanolamine,

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

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

[0187] 1,2 - Dithiohexadecatrienoic 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 comprise one or more therapeutic and / or prophylactic agents selected from vaccines or compounds capable of eliciting 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, said methods comprising administering to a mammal and / or contacting mammalian cells with a therapeutic and / or prophylactic nanoparticle composition.

[0192] In certain embodiments, the therapeutic and / or prophylactic 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 be of any size and can have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA can have a therapeutic effect when expressed in a cell.

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

[0194] The nanoparticle composition can be characterized by a variety of methods. For example, microscopy (e.g., transmission electron microscopy or scanning electron microscopy) can be used to examine the morphology and size distribution of the nanoparticle composition. Dynamic light scattering or potentiometry (e.g., potentiometric titration) can be used to measure the ζ potential. Dynamic light scattering can also be used to determine the 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 represent the uniformity of the nanoparticle composition, e.g., 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 present application further provides a method for specifically delivering a therapeutic and / or prophylactic agent to a mammalian organ, the method comprising administering to the mammal a nanoparticle composition of any of the foregoing, the administration comprising contacting the mammalian organ with the nanoparticle composition, whereby the therapeutic and / or prophylactic agent is delivered to the organ. Therapeutic and / or prophylactic 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 where the therapeutic and / or prophylactic is mRNA, when the cell contacts the nanoparticle composition, the translatable mRNA can be translated in the cell to produce the target polypeptide. However, substantially untranslatable mRNAs can also be delivered to cells. Substantially untranslatable mRNAs can be used as vaccines and / or can sequester the translation components of the cell to reduce the expression of other species in the cell.

[0198] In some embodiments, the nanoparticle composition can target a specific type or class of cells (e.g., cells of its specific organ or system). For example, a nanoparticle composition comprising a target therapy and / or prophylaxis can be specifically delivered to the liver, kidney, spleen, femur or lung of a mammal. Specific delivery to a particular class of cells, organs or their systems or groups means that a higher proportion of the nanoparticle composition comprising the therapeutic and / or prophylactic agent is delivered to the target destination (e.g., tissue) relative to other destinations, including the therapeutic and / or prophylaxis, e.g., when the nanoparticle composition is administered to the 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 blood vessels (e.g., intracoronary or intr femoral), or the 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. A compound can contain one or more chiral centers and / or double bonds and thus can exist in the form of stereoisomers, e.g., 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. Mixtures of enantiomers and stereoisomers of compounds and methods for separating them into their component enantiomers or stereoisomers are well known.

[0200] Examples

[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), and the reaction was stirred at room temperature for 3 hours while being monitored by TLC. After the reaction was complete, the reaction solution was concentrated, and compound C (5.03 g, 91.98%) was obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1).

[0206] Under an ice-water bath and under a nitrogen atmosphere, an ether solution of compound E (55 ml, 55 mmol) was added dropwise to a solution of compound D (2.00 g, 45.40 mmol) in THF (80 ml). After the addition was complete, the reaction was stirred at room temperature for 2 hours while being monitored by TLC. After the reaction was complete, 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 compound F (2.74 g, yield: 32.4%) was obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 30:1).

[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 reaction was stirred at room temperature for 3 hours while being monitored by TLC until the reaction was complete. The reaction solution was filtered, the filter cake was washed with DCM (5 ml), the filtrate was concentrated, and 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 reaction was heated to 70 °C for 20 hours while being monitored by TLC until the reaction was complete. The reaction solution was filtered, the filtrate was concentrated, and compound 1 (260.0 mg, yield: 55.3%) was obtained by silica gel column chromatography (MeOH: DCM = 20:1). 11H NMR (400 MHz, CDCl3) δ 4.95–4.80 (m, 2H), 3.69 (t, J = 6.4 Hz, 2H), 2.36–2.29 (m, 6H), 1.72–1.46 (m, 21H), 1.45–1.22 (m, 58H), 0.92 (t, J = 8.0 Hz, 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 that of Compound 1. Using propionaldehyde instead of Compound D as the raw material, Compound 2 can be obtained. 1 1H NMR (400 MHz, CDCl3) δ 4.95–4.80 (m, 2H), 3.68 (t, J = 6.4 Hz, 2H), 2.63–2.49 (m, 6H), 2.32 (q, J = 7.8 Hz, 4H), 1.73–1.47 (m, 20H), 1.43–1.27 (m, 52H), 0.91 (t, J = 8.0 Hz, 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 that of Compound 1. Using n-butanal instead of Compound D as the raw material, Compound 3 can be obtained. 1 1H NMR (400 MHz, CDCl3) δ 4.95–4.80 (m, 2H), 3.70 (t, J = 6.4 Hz, 2H), 2.34 (t, J = 7.8 Hz, 4H), 1.69–1.58 (m, 6H), 1.58–1.50 (m, 10H), 1.50–1.22 (m, 62H), 0.94 (t, J = 8.0 Hz, 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 that of Compound 1. Using n-pentanal instead of Compound D as the raw material, Compound 4 can be obtained. 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 that of Compound 1. Using acrolein instead of Compound D as the raw material, Compound 5 can be obtained. 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,6H),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 synthesis route is as follows

[0224]

[0225] Specifically, to a dichloromethane (8 ml) solution of Compound G (1.52 g, 7.80 mmol), 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. The reaction was stirred at room temperature for 3 hours, and the reaction was monitored by TLC until completion. The reaction solution was filtered, and the filter cake was washed with DCM (20 ml). The filtrate was concentrated, and silica gel column chromatography (petroleum ether:ethyl acetate = 100:1) was used to obtain Compound J (2.07 g, yield: 61.23%).

[0226] To a solution of compound J (248.8 mg, 0.66 mmol) in CPME / CH3CN (6 ml, 1V / 1V) was added compound C (300.0 mg, 0.60 mmol), K2CO3 (249.8 mg, 1.80 mmol) and KI (100.0 mg, 0.60 mmol). The mixture was heated to 70 °C and reacted for 20 h. The reaction was monitored by TLC until completion. The reaction mixture was filtered, and the filtrate was concentrated. Compound 6 (0.39 g, yield: 76.10%) was obtained by silica gel column chromatography (MeOH:DCM = 20:1). 1 1H NMR (400 MHz, CDCl3) δ 4.95–4.80 (m, 2H), 3.72 (t, J = 6.8 Hz, 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 = 8.0 Hz, 12H); MS-ESI (m / z): 851.0 (M+H) + 。

[0227] Example 7: Synthesis of compound 7

[0228]

[0229] The preparation method was the same as that of compound 6. Compound 7 could be prepared using 8-bromooctanoic acid instead of compound G as the raw material. 1 1H NMR (400 MHz, CDCl3) δ 4.95–4.80 (m, 2H), 3.70 (t, J = 6.8 Hz, 2H), 3.08 - 3.03 (m, 6H), 2.32 (t, J = 8.0 Hz, 4H), 1.94–1.86 (m, 6H), 1.67–1.48 (m, 16H), 1.41–1.30 (m, 62H), 0.92 (t, J = 8.0 Hz, 12H). MS-ESI (m / z): 879.2 (M+H) + 。

[0230] Example 8: Synthesis of compound 8

[0231]

[0232] The preparation method was the same as that of compound 1. Compound 8 could be prepared using ethanolamine and acrolein instead of compound A and compound D as the raw materials. 11H NMR (400 MHz, CDCl3) δ 5.87–5.74 (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 = 8.0 Hz, 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 that of Compound 1. Using ethanolamine, acrolein, and 8-bromo octanoic acid instead of Compound A, Compound D, and Compound G as raw materials, Compound 9 can be prepared. 1 1H NMR (400 MHz, CDCl3) δ 6.00–5.64 (m, 1H), 5.43–5.06 (m, 4H), 4.22–4.11 (m, 1H), 3.44 (t, J = 8.0 Hz, 4H), 2.36 (t, J = 7.6 Hz, 4H), 1.94–1.85 (m, 4H), 1.74–1.21 (m, 64H), 0.92 (t, J = 6.8 Hz, 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 that of Compound 1. Using 3-amino-1-propanol and acrolein instead of Compound A and Compound D as raw materials, Compound 10 can be prepared. 1 1H NMR (400 MHz, CDCl3) δ 5.88–5.74 (m, 1H), 5.31–5.14 (m, 3H), 4.97–4.84 (m, 1H), 3.91 (t, J = 5.6 Hz, 2H), 3.37–2.96 (m, 6H), 2.47–2.26 (m, 4H), 2.15–1.19 (m, 64H), 0.92 (t, J = 6.4 Hz, 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 that of Compound 1. Compound 11 can be prepared by using 4-amino-1-butanol and acrolein instead of Compound A and Compound D as raw materials. 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 synthesis route is as follows:

[0245]

[0246] Specifically, under an ice-water bath and under the protection of a nitrogen atmosphere, an ether solution of Compound E (107 ml, 107 mmol) was added dropwise to a THF (50 ml) solution of Compound K (5.00 g, 89.19 mmol). After the addition was complete, the reaction was stirred at room temperature for 2 hours, and the reaction was monitored by TLC. After the reaction was complete, saturated ammonium chloride solution (80 mL) was added to the reaction solution, and it was extracted with dichloromethane (80 ml×2), washed with saturated brine (80 ml), dried over anhydrous sodium sulfate, concentrated, and silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) was used 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 dichloromethane (30 ml) solution of Compound L (3.00 g, 15.13 mmol), and the reaction was stirred at room temperature for 3 hours, and the reaction was monitored by TLC until it was complete. The reaction solution was filtered, and the filter cake was washed with DCM (20 ml). The filtrate was concentrated, and silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) was used to obtain Compound M (5.00 g, yield: 88.03%).

[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 and reacted for 16 h. The reaction was monitored by TLC. After completion of the reaction, the reaction solution was concentrated, and compound 15 (70.0 mg, 6.60%) was obtained by silica gel column chromatography (dichloromethane:methanol = 25:1). 1 H NMR (400 MHz, CDCl3) δ 5.91–5.72 (m, 2H), 5.30–5.16 (m, 6H), 4.11–3.96 (m, 2H), 3.26–3.01 (m, 6H), 2.39 (t, J = 7.2 Hz, 4H), 2.07–1.18 (m, 48H), 0.92 (t, J = 6.4 Hz, 6H); MS-ESI (m / z): 650.7 (M+H) + 。

[0249] Example 13: Synthesis of compound 16

[0250]

[0251] Compound 16 was prepared in the same manner as compound 1, using ethanolamine, compound M, acrolein, octylmagnesium bromide, and 8-bromooctanoic acid instead of compound A, compound B, compound D, compound E, and compound G as starting materials. 1 H NMR (400 MHz, CDCl3) δ 5.83–5.70 (m, 2H), 5.26–5.11 (m, 6H), 3.68 (t, J = 5.2 Hz, 2H), 2.76 (t, J = 5.1 Hz, 2H), 2.63 (q, J = 5.2 Hz, 3H), 2.31 (q, J = 7.2 Hz, 4H), 1.60 (ddd, J = 27.1, 14.6, 7.3 Hz, 12H), 1.45–1.12 (m, 38H), 0.88 (t, J = 6.6 Hz, 6H). MS-ESI (m / z): 650.5 (M+H) + 。

[0252] Example 14: Synthesis of compound 17

[0253]

[0254] Compound 17 was prepared in the same manner as compound 1, using propanolamine, acrolein, and octylmagnesium bromide instead of compound A, compound D, and compound E as starting materials. 11H NMR (400 MHz, CDCl3) δ 5.81 (ddd, J = 17.0, 10.5, 6.3 Hz, 1H), 5.30–5.15 (m, 3H), 4.90 (p, J = 6.3 Hz, 1H), 3.84 (t, J = 5.2 Hz, 2H), 2.80 (d, J = 6.0 Hz, 2H), 2.59 (s, 3H), 2.34 (dt, J = 18.9, 7.4 Hz, 4H), 1.79 (p, J = 5.4 Hz, 2H), 1.73–1.51 (m, 14H), 1.42–1.24 (m, 46H), 0.92 (t, J = 6.8 Hz, 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 that of Compound 1. Using ethanolamine, acrolein, and octylmagnesium bromide instead of Compound A, Compound D, and Compound E as raw materials, Compound 18 can be prepared. 1 1H NMR (400 MHz, CDCl3) δ 5.77 (ddd, J = 17.0, 10.5, 6.3 Hz, 1H), 5.26–5.12 (m, 3H), 4.86 (p, J = 6.3 Hz, 1H), 3.53 (s, 1H), 2.58 (t, J = 5.4 Hz, 2H), 2.46 (dq, J = 8.3, 4.9 Hz, 4H), 2.29 (dt, J = 15.3, 7.5 Hz, 4H), 1.62 (dq, J = 15.8, 7.7 Hz, 6H), 1.54–1.42 (m, 8H), 1.38–1.18 (m, 46H), 0.88 (t, J = 6.7 Hz, 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 that of Compound 1. Using ethanolamine, acrolein, octylmagnesium bromide, and bromoacetic acid instead of Compound A, Compound D, Compound E, and Compound G as raw materials, Compound 19 can be prepared. 11H NMR (400 MHz, CDCl3) δ 5.80 (ddd, J = 17.2, 10.5, 6.7 Hz, 1H), 5.35–5.16 (m, 3H), 4.90 (p, J = 6.2 Hz, 1H), 3.58 (t, J = 5.2 Hz, 2H), 3.40 (s, 2H), 2.81 (t, J = 5.2 Hz, 2H), 2.70–2.61 (m, 2H), 2.31 (t, J = 7.5 Hz, 2H), 1.64 (d, J = 7.0 Hz, 2H), 1.57–1.46 (m, 6H), 1.37–1.26 (m, 44H), 0.92 (t, J = 6.7 Hz, 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 that of Compound 1. Using ethanolamine, acrolein, octylmagnesium bromide, and bromobutyric acid instead of Compound A, Compound D, Compound E, and Compound G as raw materials, Compound 20 can be prepared. 1 1H NMR (400 MHz, CDCl3) δ 5.80 (ddd, J = 17.2, 10.5, 6.7 Hz, 1H), 5.35–5.16 (m, 3H), 4.90 (p, J = 6.2 Hz, 1H), 3.58 (t, J = 5.2 Hz, 2H), 3.40 (s, 2H), 2.81 (t, J = 5.2 Hz, 2H), 2.70–2.61 (m, 2H), 2.31 (t, J = 7.5 Hz, 2H), 1.64 (d, J = 7.0 Hz, 2H), 1.57–1.46 (m, 6H), 1.37–1.26 (m, 44H), 0.92 (t, J = 6.7 Hz, 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 that of Compound 15. Using octylmagnesium bromide and 8-bromoheptanoic acid instead of Compound E and Compound G as raw materials, Compound 21 can be prepared. 11H NMR (400 MHz, CDCl3) δ 5.76 (ddd, J = 17.1, 10.5, 6.4 Hz, 2H), 5.24–5.18 (m, 4H), 5.15 (dd, J = 10.5, 1.3 Hz, 2H), 3.96 (t, J = 4.7 Hz, 2H), 3.10 (d, J = 5.1 Hz, 2H), 3.01 (t, J = 8.3 Hz, 4H), 2.30 (t, J = 7.4 Hz, 4H), 1.80 (t, J = 8.5 Hz, 4H), 1.61 (s, 5H), 1.50–1.07 (m, 40H), 0.87 (t, J = 6.7 Hz, 6H). MS-ESI (m / z): 650.5 (M+H) + 。

[0267] Example 19: Synthesis of Compound 23

[0268]

[0269] The preparation method was the same as that of Compound 1. Compound 23 could be prepared by using ethanolamine, acrolein, and bromobutyric acid instead of Compound A, Compound D, and Compound G as raw materials. 1 1H NMR (400 MHz, CDCl3) δ 5.31–5.16 (m, 3H), 2.59–2.49 (m, 4H), 2.34 (dt, J = 24.5, 7.4 Hz, 4H), 1.68–1.46 (m, 10H), 1.36–1.26 (m, 46H), 0.92 (t, J = 6.7 Hz, 9H). MS-ESI (m / z): 708.7 (M+H) + 。

[0270] Example 20: Synthesis of Compound 24

[0271]

[0272] The preparation method was the same as that of Compound 1. Compound 24 could be prepared by using acrolein, octylmagnesium bromide, and bromoacetic acid instead of Compound D, Compound E, and Compound G as raw materials. 1 1H NMR (400 MHz, CDCl3) δ 5.38–5.14 (m, 3H), 3.68 (t, J = 6.6 Hz, 2H), 3.36 (s, 2H), 2.65–2.56 (m, 4H), 2.31 (t, J = 7.5 Hz, 2H), 1.71–1.44 (m, 16H), 1.32 (d, J = 15.5 Hz, 44H), 0.92 (t, J = 6.7 Hz, 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 that of Compound 1. Compound 25 can be prepared by using acrolein, octylmagnesium bromide, and bromobutyric acid instead of Compound D, Compound E, and Compound G as raw materials. 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 that of Compound 1. Compound 26 can be prepared by using acrolein and bromoacetic acid instead of Compound D and Compound G as raw materials. 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 that of Compound 1. Compound 27 can be prepared by using acrolein and bromobutyric acid instead of Compound D and Compound G as raw materials. 11H NMR (400 MHz, CDCl3) δ 5.31–5.14 (m, 3H), 3.67 (t, J = 6.6 Hz, 2H), 2.51–2.39 (m, 6H), 2.33 (dt, J = 24.3, 7.5 Hz, 4H), 1.79 (p, J = 7.4 Hz, 2H), 1.67–1.27 (m, 64H), 0.91 (t, J = 6.7 Hz, 9H). MS-ESI (m / z): 764.7 (M+H) + 。

[0282] Test Example

[0283] Test Example 1: Encapsulation of Luciferase mRNA in Lipid Nanoparticles (LNP)

[0284] The stock solution of Luciferase mRNA was dispersed in 20 mM acetic acid solution (pH 5.3) to a final concentration of 200 μg / mL (aqueous phase). The lipids were mixed according to the molar ratio of the compound in the example: cholesterol: DSPC: DMG-PEG2000 = 50: 38.5: 10: 1.5 to form a lipid mixture (oil phase). By controlling the flow rates of the aqueous and oil phases through a T-mixing method, the mRNA was mixed with the lipid mixture to obtain LNP-encapsulated mRNA. The encapsulated LNP was diluted with buffer, then concentrated by ultrafiltration, and the diluent was replaced. Finally, the LNP was concentrated to a mRNA concentration of 100 μg / mL, and the pH of the LNP was adjusted to around 7 - 8. Finally, the total and free mRNA contents in the LNP were detected using a Ribogreen kit and 10% Triton as a demulsifier, and the encapsulation efficiency of the LNP was calculated. The final product of the LNP was diluted with the diluent, 1 ml was added to the particle size cell, and the particle size of the LNP was detected using a Malvern ZetaSizer instrument. The results are shown in Table 1.

[0285] Particle size, PDI, and encapsulation efficiency are all important quality attributes of lipid nanoparticles. As can be seen from the following table, the test compounds all have good encapsulation efficiency, a particle size suitable for mRNA delivery, and a narrow PDI.

[0286] Table 1: LNP Characterization Data of the Compounds in the Examples

[0287] Number Particle size (nm) PDI Entrapment efficiency (%) 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: Test of Intravenous Injection Delivery Effect

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

[0290] 2. Inject the loaded LNP formulation into Balb / c mice via intravenous injection 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. Fluorescence expression intensity detection: Inject sodium D-luciferin (dose: 150 mg / kg) into the peritoneal cavity of each mouse 10 minutes before detection. Then anesthetize the mouse with isoflurane and place it in the IVIS instrument, and select bioluminescence for detection.

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

[0294] The delivery and expression effects of the compound LNP of the present invention are as Figure 1 shown. Take the expression level of the MC3 group as the reference. The data shows that among each group, the expressions of the compound 1 group and the compound 2 group are slightly higher than that of the MC3 group, the expressions of the compound 4 group, the compound 6 group, the compound 16 group, and the compound 21 group are lower than that of the MC3 group, while the expressions of the compound 5 group, the compound 8-11 groups, and the compound 17-18 groups are significantly higher than that of the MC3 group, and the expression level is two to four times that of the MC3 group.

[0295] Experimental Example 3: Test of intramuscular injection delivery effect

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

[0297] 2. Inject the loaded LNP formulation into Balb / c mice via intramuscular injection, 50 μl for each mouse, at a dose of 5 μg / mouse, with 5 mice in each group.

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

[0299] 4. Fluorescence expression intensity detection: Inject sodium D-luciferin (dose: 150 mg / kg) into the peritoneal cavity of each mouse 10 minutes before detection. Then anesthetize the mouse with isoflurane and place it in the IVIS instrument, select bioluminescence for detection, and after the detection is completed, analyze the fluorescence intensity at the intramuscular injection site.

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

[0301] According to Figure 2As can be seen from the results, compounds 8, 10, and 11 can also achieve good expression effects through intramuscular injection, all of which are significantly better than the positive reference group MC3, and the expression advantage of the compound 8 group is the most obvious.

[0302] Although the above combination describes the implementation schemes of the present application, the present application is not limited to the above specific implementation schemes and application fields. The above specific implementation schemes are merely illustrative and guiding, rather than restrictive. Under the inspiration of this specification and without departing from the scope protected by the claims of the present application, those of ordinary skill in the art can also make many forms, all of which are within the scope of protection of the present application.

Claims

1. A compound of formula (I), or a salt or stereoisomer thereof, wherein R1 is vinyl, wherein, R2 is a straight-chain alkyl group having 7 to 11 carbon atoms, and R3 and R4 are each independently a straight-chain alkyl group having 6 to 9 carbon atoms; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, provided that X1 and Y1, X2 and Y2 are not simultaneously C=O or O; m is selected from 3, 4, 5, 6, 7, 8; n is selected from 3, 4, 5, 6, 7, 8; o is selected from 2, 3, 4, 5, 6.

2. The compound or its salt or its stereoisomer according to claim 1, wherein, R2 is a straight-chain alkyl group having 8 or 10 carbon atoms.

3. The compound or its salt or its stereoisomer according to claim 1, wherein, R3 and R4 are each independently a straight-chain alkyl group having 8 carbon atoms.

4. The compound or its salt or its stereoisomer according to claim 1, wherein, m is 5 or 7.

5. The compound or its salt or its stereoisomer according to claim 1, wherein, n is selected from 4, 5, 6, 7.

6. The compound or its salt or its stereoisomer according to claim 5, wherein, n is 7.

7. The compound or its salt or its stereoisomer according to claim 1, wherein, Both X1 and X2 are C=O, both Y1 and Y2 are O, or X1 is C=O, Y1 is O, X2 is O, Y2 is C=O, 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.

8. The compound or its salt or its stereoisomer according to claim 1, wherein, m is selected from 5, 6, 7; n is selected from 5, 6, 7; both X1 and X2 are C=O, both Y1 and Y2 are O, or X1 is C=O, Y1 is O, X2 is O, Y2 is C=O, 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.

9. The compound or its salt or its stereoisomer according to claim 1, wherein, The compound of formula (I) is selected from Compound 5 Compound 8 Compound 9 Compound 10 Compound 11 Compound 12 Compound 13 Compound 14 Compound 17 Compound 18 Compound 19 Compound 20 Compound 22 Compound 23 Compound 24 Compound 25 Compound 26 Compound 27 10. A nanoparticle composition comprising a lipid component, said lipid component comprising the compound or a salt or stereoisomer thereof according to any one of claims 1-9.

11. The nanoparticle composition according to claim 10, wherein said lipid component further comprises a phospholipid.

12. The nanoparticle composition according to claim 11, wherein said phospholipid is selected from one or more of the following compounds: Dilauroyl phosphatidylcholine (DLPC), Dimyristoyl phosphatidylcholine (DMPC), Dioleoyl phosphatidylcholine (DOPC), Dipalmitoyl phosphatidylcholine (DPPC), Distearoyl phosphatidylcholine (DSPC), Dioleoyl phosphatidylcholine (DUPC), Palmitoyl oleoyl phosphatidylcholine (POPC), 1,2-Di-O-octadecyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-Oleoyl-2-cholesteryl dimethyl succinate-sn-glycero-3-phosphocholine (OChemsPC), 1-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-Divinylol-sn-glycero-3-phosphoethanolamine, 1,2-Divinyl-sn-glycero-3-phosphoethanolamine, 1,2-Diaryl-sn-glycero-3-phosphoethanolamine, 1,2-Dithiohexadecenoic acid-sn-glycero-3-phosphoethanolamine, Sodium 1,2-dioleoyl-sn-glycero-3-phosphate-(1-glycerol) (DOPG) or sphingomyelin.

13. The nanoparticle composition according to claim 11, wherein the phospholipid is DOPE.

14. The nanoparticle composition according to claim 11, wherein the phospholipid is DSPC.

15. The nanoparticle composition according to claim 11, wherein the lipid component further comprises a structural lipid.

16. The nanoparticle composition according to claim 15, wherein the structural lipid is selected from one or more of cholesterol, coprostanol, sitosterol, ergosterol, stigmasterol.

17. The nanoparticle composition according to claim 15, wherein the structural lipid is cholesterol.

18. The nanoparticle composition according to claim 15, wherein the lipid component further comprises a PEG lipid.

19. The nanoparticle composition according to claim 18, 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.

20. The nanoparticle composition according to claim 18, wherein the lipid component further comprises a cationic and / or ionizable lipid.

21. The nanoparticle composition according to claim 20, further comprising a therapeutic agent and / or a prophylactic agent selected from a vaccine or a compound capable of eliciting an immune response, nucleic acid.

22. The nanoparticle composition according to claim 21, wherein the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA, or mRNA.

23. The nanoparticle composition according to claim 21 or 22, wherein the encapsulation rate of the therapeutic agent and / or prophylactic agent is ≥ 50%; or ≥ 80%; or ≥ 90%.

24. The nanoparticle composition according to any one of claims 10-22, wherein the average particle size of the nanoparticle composition is 50 nm - 110 nm.

25. The nanoparticle composition according to any one of claims 10-22, wherein the dispersity index of the nanoparticle composition is 0.04 - 0.

20.

26. Use of the compound according to any one of claims 1-9 in the preparation of a lipid nanoparticle composition.

27. A pharmaceutical composition comprising the nanoparticle composition according to any one of claims 10-25 and a pharmaceutically acceptable carrier.

28. Use of the nanoparticle composition according to any one of claims 10-25 or the pharmaceutical composition according to claim 27 in the preparation of a drug for delivering a therapeutic agent and / or a prophylactic agent to mammalian cells, the administration of the drug comprising contacting the cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic agent and / or prophylactic agent to the cells.

29. Use according to claim 28, wherein the mammalian cell is in a mammal.

30. Method according to claim 28 or 29, wherein the mammal is a human.

31. Method according to claim 28 or 29, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally or by inhalation.

32. Use of the nanoparticle composition according to any one of claims 10 - 25 or the pharmaceutical composition according to claim 27 in the preparation of a medicament for producing a polypeptide of interest in mammalian cells, the administration of the medicament comprising contacting the cells with the nanoparticle composition or the pharmaceutical composition to deliver a therapeutic agent and / or a prophylactic agent to the cells, wherein the therapeutic agent and / or the prophylactic agent is mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cells to produce the polypeptide of interest.

33. Use according to claim 32, wherein the mammalian cell is in a mammal.

34. Use according to any one of claims 32 or 33, wherein the mammalian cell is a human.

35. Use according to any one of claims 32 or 33, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally or by inhalation.

36. Use of the nanoparticle composition according to any one of claims 10 - 25 or the pharmaceutical composition according to claim 27 in the preparation of a medicament for treating a disease or disorder in a mammal, the medicament being administered to the mammal in a therapeutically effective amount.

37. Use according to claim 36, wherein the disease or disorder is characterized by a dysfunctional or abnormal protein or polypeptide activity.

38. Use according to claim 36 or 37, wherein the disease or disorder is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases or metabolic diseases.

39. Use according to claim 36 or 37, wherein the mammal is a human.

40. Use according to claim 36 or 37, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally or by inhalation.

41. Use of the nanoparticle composition according to any one of claims 10 - 25 or the pharmaceutical composition according to claim 27 in the preparation of a medicament for specifically delivering a therapeutic agent and / or a prophylactic agent to a mammalian organ, the administration of the medicament comprising contacting the mammalian organ with the nanoparticle composition, whereby the therapeutic agent and / or the prophylactic agent is delivered to the organ.

42. Use according to claim 41, wherein the mammal is a human.

43. Use according to claim 41 or 42, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally or by inhalation.

44. Use according to claim 41 or 42, wherein the mammal is pretreated 24 hours or less before the contacting or administration step.

45. The use according to claim 41 or 42, wherein the mammal is pretreated about one hour before the contacting or administering step.

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