Lipid Compounds and Lipid Nanoparticle Compositions

Abstract

The present application discloses a compound of formula (I) and salts and stereoisomers thereof, wherein each variable is as defined herein. The present application also discloses nanoparticle compositions comprising the compound or a salt or stereoisomer thereof, and uses of the nanoparticle compositions for delivering active agents. JPEG2026502084000140.jpg51128

Description

[Technical Field]

[0001] This application relates to the field of biotechnology, and in particular to lipid compounds and lipid nanoparticle compositions. In particular, this application relates to methods for delivering active agents, such as nucleic acids, and lipid compounds and lipid nanoparticle compositions for delivering the same. [Background technology]

[0002] Efficient targeted delivery of biologically active substances (e.g., small molecule drugs, proteins, and nucleic acids) remains a continuing medical challenge. The key to successful gene therapy is the safe and effective in vivo delivery of therapeutic agents to target cells via vectors. Nucleic acids are relatively unstable and have poor cellular permeability, making their delivery into cells challenging. Therefore, there is a need to develop methods and compositions that facilitate the delivery of therapeutic and / or prophylactic agents, such as nucleic acids, into cells. Gene therapy vectors are classified into viral and non-viral vectors. Viral vectors have been used as highly efficient delivery systems for target gene transfection and therapeutic purposes. However, they suffer from issues such as the presence of immunogenic viral proteins, limited target gene payload, and high cost. Lipid nanoparticles (LNPs), a non-viral vector, have been widely explored due to their advantages, such as good in vitro stability, in vivo degradability, safety, and reliability, and have been widely applied in gene therapy research for congenital and acquired gene deficiencies.

[0003] Lipid-containing nanoparticles (lipid nanoparticles), liposomes, and lipid complexes have been demonstrated to be effective transport vectors for the delivery of biologically active substances, such as small molecule drugs, proteins, and nucleic acids, into and / or within cells. LNPs are small vesicles formed from one or more lipid components that are effective in compacting and delivering various nucleic acid molecules, from DNA and RNA to chromosomes and even cells. LNPs are advantageous for large-scale production due to their defined assembly scheme and properties, such as the ability to easily modify them for targeting with ligands.

[0004] LNPs typically contain one or more cationic lipids and / or amino (pH-ionizable) lipids, phospholipids, structural lipids (such as cholesterol), and / or polyethylene glycol-containing lipids (PEG-lipids). Cationic lipids and / or pH-responsive lipids (including, for example, amine-containing lipids) can be readily protonated.

[0005] Lipid nanoparticle (LNP) formulations represent a major advancement in the field of nucleic acid delivery. Onpattro™ is an early example of a lipid nanoparticle product approved for clinical use. Onpattro™ is a lipid nanoparticle-based small interfering RNA (siRNA) pharmaceutical formulation for the treatment of polyneuropathy caused by hereditary transthyretin amyloidosis. The success of this LNP delivery system paves the way for the clinical development of a lead LNP-based mRNA COVID-19 vaccine.

[0006] The Onpattro™ LNP formulation consists of four major lipid components: the pH-responsive amino lipid MC3, distearoylphosphatidylcholine (DSPC), cholesterol, and polyethylene glycol-conjugated lipid (PEG-lipid) in molar amounts of 50 / 10 / 38.5 / 1.5, respectively. Onpattro™ is still considered the gold standard for comparison in LNP research.

[0007] Although LNP-mediated nucleic acid delivery is well-established, it is well known that the Onpattro™ formulation accumulates primarily in liver tissue. The clinical utility of this delivery system would be greatly expanded if LNPs could also accumulate in organs and tissues other than the liver. Summary of the Invention

[0008] Based on the above, the present application discloses lipid compounds and lipid nanoparticle compositions comprising such compounds, which have advantages such as high encapsulation efficiency, low toxicity, high expressibility, suitability for vaccine or mRNA delivery, extrahepatic delivery, etc.

[0009] Specifically, the present application includes the following embodiments (Part I): 1. Formula (I),

[0010] [ka] [In the formula, R1 is

[0011] [ka] (In the formula, R 1a and R 1b are independently selected from H or C1-C6 alkyl, and R2 and R3 are independently selected from H or C1-C6 alkyl, or R1 and R2 together with the carbon atoms to which they are attached form a substituted or unsubstituted 5- to 7-membered nitrogen-containing heterocycle, and R3 is selected from H or C1-C6 alkyl, or R1 and R3 together with the carbon atoms to which they are attached form a ring A which is a substituted or unsubstituted 5-7 membered carbocyclic or heterocyclic ring, and R2 is selected from H or C1-C6 alkyl; R4, R5, and R6 are independently selected from C1-C14 alkyl; X and Y are independently selected from O or S; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; m and n are independently selected from 0, 1, or 2, with the proviso that m and n are not both 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; q is selected from 0, 1, 2, 3, 4, 5 or 6, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 2. The compound of formula (I) is represented by formula (IA):

[0012] [ka]

[0023] Item 1. The compound according to item 1, wherein q is selected from 1, 2, 3, or 4, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 3. The compound according to item 1 or 2, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, wherein R4 is C9, C10, C11, or C12 straight-chain alkyl; and R5 and R6 are each independently selected from C6, C7, C8, or C9 straight-chain alkyl. 4. The compound according to any one of items 1 to 3, wherein o is 5 or 6, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 5. The compound according to any one of items 1 to 4, wherein p is 7 or 8, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 6. The compound according to any one of items 1 to 5, wherein X and Y are both O, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 7. n is 1 and m is 0; or n is 1 and m is 1; or n is 0 and m is 1; For example, n is 1 and m is 0; for example, n is 1 and m is 1; the compound according to any one of items 1 to 6, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 8. The compound of formula (I) has the following structure:

[0013] [ka] Item 8. The compound according to any one of items 1 to 7, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, selected from: 9. The compound of formula (I) has the following structure:

[0014] [ka] Item 9. The compound according to any one of items 1 to 8, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, selected from: 10. The compound of formula (I) has the following structure:

[0015] [ka] Item 10. The compound according to any one of items 1 to 9, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, selected from: 11. C1-C6 alkyl is independently selected from -CH3, -CH2CH3, or -CH(CH3)2; For example, the compound according to any one of items 1 to 10, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, wherein C1-C6 alkyl is independently selected from -CH3. 12. R 1a and R 1b are independently selected from -CH3; R2 and R3 are independently selected from H; R4 is a C10, C11, or C12 straight chain alkyl; R5 and R6 are the same and are selected from C6, C7 and C8 straight chain alkyl; X and Y are independently selected from O; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; m is 0 and n is 1; o is 5 or 6; p is 7 or 8, for example, p is 7; q is 2, 3 or 4, for example, q is 2 or 3, for example, q is 2; For example, p is 7 and q is 2 or 3; For example, the compound according to any one of items 1 to 11, wherein p is 7 and q is 2, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 13. The compound according to any one of items 1 to 12, wherein X1 and X2 are both C=O, and Y1 and Y2 are both O, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 14. The compound according to any one of items 1 to 12, wherein X1 is O, Y1 is C=O, X2 is C=O, and Y2 is O, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 15. The compound according to any one of items 1 to 12, wherein X1 is C=O, Y1 is O, X2 is O, and Y2 is C=O; or X1 and X2 are both O, and Y1 and Y2 are both C=O, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 16. The compound according to any one of items 1 to 15, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, wherein R4 is C10-C11 straight chain alkyl. 17. The compound according to any one of paragraphs 1 to 16, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, wherein R5 and R6 are both C8 straight chain alkyl. 18. o is 5, R4 is a C11 straight chain alkyl, p is 7, and R5 and R6 are both C8 straight chain alkyl, or Item 18. The compound according to any one of items 1 to 17, wherein o is 6, R4 is C10 straight chain alkyl, p is 7, and R5 and R6 are both C8 straight chain alkyl, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof. 19.

[0016] [ka]

[0017] [ka]

[0018] [ka] Item 1, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, selected from: 20.

[0019] [ka]

[0020] [ka] Item 1, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, selected from: 21. A process for preparing a compound according to any one of items 1 to 20, wherein compound (IA) is obtained from compound J as a starting material,

[0021] [ka] wherein Z1 is selected from OH, Z2 and Z3 are each independently selected from OH or SH, and Z4 is selected from OMs (Ms is methylsulfonyl); X, Y, X1, X2, Y1, Y2, R 1a , R 1b , R2, R3, R4, R5, R6, m, n, o, p and q are as defined in any one of items 1 to 20; Compound J is reacted with compound K in a benzene-based solution of pyridinium 4-methylbenzenesulfonate to provide compound L, and the benzene-based solvent used in the benzene solution is preferably toluene; The compound L is reacted in a solution of (CH3SO2)2O in a haloalkane to provide the compound M, the haloalkane being preferably DCM, dichloroethane or chloroform; The process wherein compound M is reacted in a polar aprotic solvent to form compound IA, the polar aprotic solvent preferably being THF. 22. Compound J is obtained from compound H and compound C as starting materials,

[0022] [ka] wherein X is selected from halogen; X1, X2, R4, R5, R6, o and p are as defined in any one of items 1 to 20; The compound H is reacted with the compound C in the presence of a carbonate, NBuI, and a polar aprotic solvent to provide the compound I, wherein the carbonate is preferably CsCO or KCO; the polar aprotic solvent is preferably DMF or DMA; Item 22. The preparation method according to item 21, wherein compound I is reacted at pH 1-5 to provide compound J, preferably the pH is adjusted by hydrochloric acid solution, and the reactant is a haloalkane, preferably DCM, dichloroethane or chloroform. 23. Compound F reacts with compound G to provide compound H;

[0023] [ka] wherein X is selected from halogen; X1, Y1, R4 and o are as defined in any one of items 1 to 20; Item 22. The process according to item 22, wherein compound F is reacted with compound G in the presence of a carbonate, NBuI and a polar aprotic solvent to provide compound H, wherein the carbonate is preferably CsCO or KCO; and the polar aprotic solvent is preferably DMF or DMA. 24. A nanoparticle composition comprising a lipid component containing the compound according to any one of items 1 to 20, or a salt (particularly a pharmaceutically acceptable salt) or stereoisomer thereof, preferably the nanoparticle composition being a lipid nanoparticle (LNP). 25. The lipid component further comprises a phospholipid, a structured lipid, and / or a PEG lipid; The phospholipid preferably comprises the following compound: Dilauryl lecithin (DLPC), Dimyristoylphosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dioleoylphosphatidylcholine (DUPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 lyso-PC), 1,2-divinyl-sn-glycero-3-phosphocholine, 1,2-diarylacyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-diethenol-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexaenoate-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphate-(1-glycerol) sodium salt (DOPG) or sphingomyelin; For example, the phospholipid is DOPE or DSPC; the structural lipid is preferably selected from one or more of cholesterol, coprosterol, sitosterol, ergosterol and stigmasterol; for example, the structural lipid is cholesterol; and / or Item 25. The nanoparticle composition according to Item 24, wherein the PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol. 26. The nanoparticle composition according to item 24 or 25, wherein the lipid component further comprises a cationic lipid and / or a pH-responsive lipid. 27. The method further comprises a therapeutic and / or prophylactic agent, wherein the therapeutic and / or prophylactic agent is selected from a vaccine, or a compound capable of inducing an immune response, and / or a nucleic acid; the nucleic acid is preferably RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA, or mRNA; For example, the nanoparticle composition according to any one of Items 24 to 26, wherein the therapeutic agent and / or the prophylactic agent is mRNA. 28. the encapsulation efficiency of the therapeutic agent and / or the prophylactic agent is 50% or more; or 80% or more; or 90% or more; and / or the nanoparticle composition has an average particle size of 50 nm to 110 nm; and / or Item 28. The nanoparticle composition according to any one of Items 24 to 27, wherein the nanoparticle composition has a polydispersity index of 0.04 to 0.20. 29. Use of the compound according to any one of items 1 to 20, or a salt thereof (particularly a pharmaceutically acceptable salt) or a stereoisomer thereof in the preparation of a lipid nanoparticle composition. 30. A pharmaceutical composition comprising the nanoparticle composition according to any one of items 24 to 28 and a pharmaceutically acceptable excipient. 31. A method for delivering a therapeutic agent and / or prophylactic agent to mammalian cells, comprising administering to a subject the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, wherein the administration comprises contacting the cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic agent and / or prophylactic agent to the cells; for example, the mammalian cells are in a mammal; for example, the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 32. A method for producing a polypeptide of interest in a mammalian cell, comprising contacting the cell with the nanoparticle composition of any one of claims 24 to 28 or the pharmaceutical composition of claim 30 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, and the mRNA is capable of being translated in the cell to produce the polypeptide of interest; for example, the mammalian cell is in a mammal; for example, the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 33. A method for treating or preventing a disease or condition in a mammal such as a human, comprising administering to the mammal a therapeutically or prophylactically effective amount of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30; for example, the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity; for example, the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 34. A method for specifically delivering a therapeutic agent and / or prophylactic agent to an organ of a mammal, comprising administering to the mammal the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, wherein the administration comprises contacting the organ of the mammal with the nanoparticle composition or the pharmaceutical composition, thereby delivering a therapeutic agent and / or prophylactic agent to the organ, wherein the mammal is, for example, a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation; for example, the mammal is pretreated 24 hours or less before the contacting or administering step; for example, about 1 hour before the contacting or administering step. 35. A method for delivering a therapeutic agent and / or a prophylactic agent to a patient, the method comprising administering to a patient in need thereof the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30. 36. A method for introducing nucleic acid into cells, comprising contacting the cells with the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30. 37. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, for use as a medicine. 38. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in delivering a therapeutic and / or prophylactic agent to mammalian cells, wherein, for example, the mammalian cells are in a mammal; for example, the mammal is a human; and the nanoparticle composition or pharmaceutical composition is administered, for example, intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 39. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, which is used to produce a polypeptide of interest in a mammalian cell, wherein the composition comprises a therapeutic and / or prophylactic agent that is an mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest; for example, the mammalian cell is in a mammal; for example, the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 40. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in treating or preventing a disease or condition in a mammal such as a human. 41. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in treating or preventing a disease or condition in a mammal such as a human, wherein the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity; for example, the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, and metabolic diseases; for example, the nanoparticle composition or pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 42. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in the specific delivery of a therapeutic and / or prophylactic agent to an organ of a mammal, wherein the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 43. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in the specific delivery of a therapeutic and / or prophylactic agent to an organ of a mammal, for example, wherein the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation, and the mammal is pretreated 24 hours or less before the contacting or administering step; for example, about 1 hour before the contacting or administering step. 44. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, for delivering a therapeutic agent and / or a prophylactic agent to a patient. 45. The nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30, for introducing a nucleic acid into a cell. 46. ​​Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the manufacture of a medicament for delivering a therapeutic and / or prophylactic agent to mammalian cells, wherein, for example, the mammalian cells are in a mammal; for example, the mammal is a human; and for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 47. Use of the nanoparticle composition of any one of paragraphs 24 to 28 or the pharmaceutical composition of paragraph 30 in the manufacture of a medicament for producing a polypeptide of interest in a mammalian cell, wherein the medicament comprises a therapeutic and / or prophylactic agent that is an mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest; for example, the mammalian cell is in a mammal; for example, the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 48. Use of the nanoparticle composition according to any one of paragraphs 24 to 28 or the pharmaceutical composition according to paragraph 30 in the manufacture of a medicament for the treatment or prevention of a disease or condition in a mammal, such as a human. 49. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the manufacture of a medicament for the treatment or prevention of a disease or condition in a mammal, for example, a human, wherein the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity; for example, the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, or metabolic diseases; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 50. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the manufacture of a medicament for specific delivery of a therapeutic and / or prophylactic agent to an organ of a mammal, wherein the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 51. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the manufacture of a medicament for specific delivery of a therapeutic and / or prophylactic agent to an organ of a mammal, for example, wherein the mammal is a human; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation, and the mammal is pretreated 24 hours or less before the contacting or administering step; for example, about 1 hour before the contacting or administering step. 52. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the manufacture of a medicament for delivering a therapeutic and / or prophylactic agent to a patient. 53. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 in the preparation of a product for introducing nucleic acid into cells. 54. Use of the nanoparticle composition according to any one of items 24 to 28 or the pharmaceutical composition according to item 30 for use in treating or preventing a disease or condition in a mammal such as a human; for example, the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity; for example, the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, or metabolic diseases; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

[0024] The present application further includes the following embodiments (Part II): 1. Formula (I),

[0025] [ka] [Wherein R1 is

[0026] [ka] (In the formula, R 1a , R 1b are independently selected from H and C1-C6 alkyl, and R2, R3 are independently selected from H or C1-C6 alkyl, or R1 and R2 together with the carbon atoms to which they are attached form a substituted or unsubstituted 5- to 7-membered nitrogen-containing heterocycle, and R3 is selected from H or C1-C6 alkyl, or R1 and R3 together with the carbon atoms to which they are attached form a ring A which is a substituted or unsubstituted 5-7 membered carbocyclic or heterocyclic ring, and R2 is selected from H or C1-C6 alkyl; R4, R5, and R6 are independently selected from C1-C14 alkyl; X and Y are independently selected from O or S; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; m and n are independently selected from 0, 1, or 2, with the proviso that m and n are not both 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and q is selected from 0, 1, 2, 3, 4, 5, or 6, or a salt or isomer thereof. 2. The compound of formula (I) is represented by formula (IA):

[0027] [ka] Item 2. The compound according to item 1, wherein q is selected from 0, 1, 2, 3, and 4. 3. The compound of formula (I) is represented by formula (IB):

[0028] [ka] wherein R7 is selected from H or C1-C6 alkyl; Item 3. The compound according to item 1, wherein r is selected from 0, 1, 2, or 3. 4. The compound of formula (I) is represented by formula (IC) or (ID),

[0029] [ka] wherein ring A is substituted with one or more R8, and R8 is

[0030] [ka] (In the formula, R 8a and R 8bare independently selected from H or C1-C6 alkyl, and s is selected from 0, 1, 2, 3, 4, 5, or 6. 5. The ring A is

[0031] [ka] Item 5. The compound according to item 4, selected from: 6. The compound according to any one of items 1 to 5, wherein R4 is C10-C11 alkyl. 7. The compound according to item 6, wherein R4 is a C10-C11 linear alkyl. 8. The compound according to any one of items 1 to 7, wherein R5 and R6 are C8 alkyl. 9. The compound according to item 8, wherein R5 and R6 are C8 straight chain alkyl. 10. The compound according to any one of items 1 to 9, wherein o is 5 or 6. 11. The compound according to any one of items 1 to 10, wherein p is 7 or 8. 12. o is 5, R4 is C11 straight chain alkyl, p is 7, R5 and R6 are C8 straight chain alkyl, or Item 12. The compound according to any one of items 1 to 11, wherein o is 6, R4 is C10 straight chain alkyl, p is 7, and R5 and R6 are C8 straight chain alkyl. 13. The compound according to any one of items 1 to 12, wherein X1 and X2 are C=O, and Y1 and Y2 are O. 14. The compound according to any one of items 1 to 12, wherein X1 is O, Y1 is C=O, X2 is C=O, and Y2 is O. 15. The compound according to any one of items 1 to 12, wherein X1 is C=O, Y1 is O, X2 is O, and Y2 is C=O. 16. The compound according to any one of items 1 to 12, wherein X1 and X2 are O, and Y1 and Y2 are C=O. 17. The compound according to any one of items 1 to 16, wherein X and Y are both O. 18. The compound according to any one of items 1 to 16, wherein n is 0 and m is 1, or n is 1 and m is 0, or n is 1 and m is 1. 19. The compound according to any one of items 1 to 18, wherein the C1-C6 alkyl is selected from -CH3, -CH2CH3, or -CH(CH3)2. 20. The compound of formula (I)

[0032] [ka]

[0033] [ka]

[0034] [ka] The compound according to item 1, selected from: 21. A method for preparing a compound according to any one of items 1 to 20, wherein compound (IA) is obtained from compound J as a starting material,

[0035] [ka] wherein Z1 is selected from OH, Z2 and Z3 are each independently selected from OH or SH, and Z4 is selected from OMs; X and Y are independently selected from O or S; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; R 1a and R 1b are independently selected from H or C1-C6 alkyl; R2 and R3 are independently selected from H or C1-C6 alkyl; R4, R5, and R6 are independently selected from C1-C14 alkyl; m and n are independently selected from 0, 1, and 2, with the proviso that m and n are not both 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; q is selected from 0, 1, 2, 3, 4, 5 or 6; The compound J is reacted with the compound K in a benzene-based solution of Tso-Py to provide the compound L, and the benzene-based solvent used is preferably toluene; The compound L is reacted in a solution of (CH3SO2)2O in a haloalkane to provide the compound M, the haloalkane being preferably DCM, dichloroethane or chloroform; The method wherein compound M is reacted in a polar aprotic solvent to produce compound IA, the polar aprotic solvent being preferably THF. 22. Compound J is obtained from compound H and compound C as starting materials,

[0036] [ka] X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; X is selected from halogens; R4, R5, and R6 are independently selected from C1-C14 alkyl; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; The compound H is reacted with the compound C in the presence of a carbonate, NBuI, and a polar aprotic solvent to provide the compound I, wherein the carbonate is preferably CsCO and KCO; the polar aprotic solvent is preferably DMF or DMA; Item 22. The preparation method according to item 21, wherein compound I is reacted at pH 1-5 to provide compound J, preferably the pH is adjusted by hydrochloric acid solution, and the reaction solvent is a haloalkane, preferably DCM, dichloroethane or chloroform. 23. Compound F reacts with compound G to provide compound H;

[0037] [ka] wherein X1 and Y1 are independently selected from C=O or O; X is selected from halogens; R4 is selected from C1-C14 alkyl; o is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; Item 22. The preparation method according to item 22, wherein compound F is reacted with compound G in the presence of a carbonate, NBuI and a polar aprotic solvent to provide compound H, wherein the carbonate is preferably CsCO, KCO; and the polar aprotic solvent is preferably DMF or DMA. 24. A nanoparticle composition comprising a lipid component containing the compound according to any one of items 1 to 20. 25. The nanoparticle composition of claim 24, wherein the lipid component further comprises a phospholipid. 26. The phospholipid is selected from the group consisting of the following compounds: Dilauryl lecithin (DLPC), Dimyristoylphosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dioleoylphosphatidylcholine (DUPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 lyso-PC), 1,2-divinyl-sn-glycero-3-phosphocholine, 1,2-diarylacyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-diethenol-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexa-enoate-sn-glycero-3-phosphoethanolamine, Item 26. The nanoparticle composition according to item 25, wherein the nanoparticle is selected from one or more of 1,2-dioleoyl-sn-glycero-3-phosphate-(1-glycerol) sodium salt (DOPG) or sphingomyelin. 27. The nanoparticle composition according to paragraph 25, wherein the phospholipid is DOPE. 28. The nanoparticle composition according to item 25, wherein the phospholipid is DSPC. 29. The nanoparticle composition according to any one of items 24 to 28, wherein the lipid component further comprises a structured lipid. 30. The nanoparticle composition of paragraph 29, wherein the structural lipid is selected from one or more of cholesterol, coprosterol, sitosterol, ergosterol, and stigmasterol. 31. The nanoparticle composition according to paragraph 29, wherein the structural lipid is cholesterol. 32. The nanoparticle composition according to any one of items 24 to 31, wherein the lipid component further comprises a PEG lipid. 33. The nanoparticle composition of paragraph 32, 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. 34. The nanoparticle composition according to any one of items 24 to 33, wherein the lipid component further comprises a cationic lipid and / or a pH-responsive lipid. 35. The vaccine, or a compound capable of inducing an immune response, further comprising a therapeutic and / or prophylactic agent selected from a nucleic acid; Preferably, the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA, and mRNA. 36. The nanoparticle composition according to any one of items 24 to 35, wherein the encapsulation efficiency of the therapeutic agent and / or the prophylactic agent is 50% or more; or 80% or more; or 90% or more. 37. The nanoparticle composition according to any one of items 24 to 36, wherein the nanoparticle composition has an average particle size of 50 nm to 110 nm. 38. The nanoparticle composition according to any one of items 24 to 36, wherein the nanoparticle composition has a polydispersity index of 0.04 to 0.20. 39. Use of a compound according to any one of paragraphs 1 to 20 in the preparation of a lipid nanoparticle composition. 40. A pharmaceutical composition comprising the nanoparticle composition according to any one of items 24 to 38 and a pharmaceutically acceptable carrier. 41. A method for delivering a therapeutic agent and / or prophylactic agent to mammalian cells, comprising administering to a subject the nanoparticle composition according to any one of items 24 to 38 or the pharmaceutical composition according to item 40, wherein the administration comprises contacting cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic agent and / or prophylactic agent to the cells. 42. The method of paragraph 41, wherein the mammalian cell is in a mammal. 43. The method of paragraph 41 or 42, wherein the mammal is a human. 44. The method of any one of paragraphs 41 to 43, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 45. A method for producing a polypeptide of interest in mammalian cells, comprising contacting the cells with the nanoparticle composition of any one of claims 24 to 38 or the pharmaceutical composition of claim 40 to deliver a therapeutic and / or prophylactic agent to the cells, wherein the therapeutic and / or prophylactic agent is mRNA encoding the polypeptide of interest, and the mRNA can be translated in the cells to produce the polypeptide of interest. 46. ​​The method of paragraph 45, wherein the mammalian cell is in a mammal. 47. The method of paragraph 45 or 46, wherein the mammal is a human. 48. The method according to any one of paragraphs 45 to 47, wherein the nanoparticle composition or pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 49. A method for treating a disease or condition in a mammal, comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition described in any one of items 24 to 38 or the pharmaceutical composition described in item 40. 50. The method of paragraph 49, wherein the disease or condition is characterized by dysfunction or abnormal activity of a protein or polypeptide. 51. The method of paragraph 49 or 50, wherein the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases or metabolic diseases. 52. The method of any one of paragraphs 49 to 51, wherein the mammal is a human. 53. The method according to any one of paragraphs 49 to 52, wherein the nanoparticle composition or pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 54. A method for specifically delivering a therapeutic agent and / or prophylactic agent to an organ of a mammal, comprising administering to the mammal the nanoparticle composition described in any one of items 24 to 38 or the pharmaceutical composition described in item 40, wherein the administration comprises contacting the organ of the mammal with the nanoparticle composition or the pharmaceutical composition, thereby delivering the therapeutic agent and / or prophylactic agent to the organ. 55. The method of paragraph 54, wherein the mammal is a human. 56. The method of paragraph 54 or 55, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation. 57. The method of any one of paragraphs 54 to 56, wherein the mammal is pretreated 24 hours or less prior to the contacting or administering step. 58. The method of any one of paragraphs 54 to 57, wherein the mammal is pretreated about 1 hour prior to the contacting or administering step. [Effects of the Invention]

[0038] The compounds of the present application can be used to prepare lipid nanoparticles, and nanoparticle compositions containing the compounds of the present application can encapsulate and deliver therapeutic / prophylactic agents, safely deliver the therapeutic / prophylactic agents to target sites, achieve high expression, and exert the effects of the therapeutic / prophylactic agents.

[0039] The lipid nanoparticles prepared in this application have the advantages of small average particle size, high encapsulation efficiency, and low toxicity, and are expected to be widely applied in the field of drug delivery.

[0040] This study reports the first synthetic method for cationic lipids with asymmetric tails and containing acetal structures. First, two different bromo-substituted tail compounds were synthesized. Subsequently, nucleophilic substitution reactions with tosyl isocyanide were carried out sequentially under different alkaline conditions to obtain the tosyl isocyanide-substituted double-long-chain compound. Finally, acid hydrolysis afforded the double-tail carbon compound as a key intermediate. DETAILED DESCRIPTION OF THE INVENTION

[0041] In the following description of exemplary embodiments of the present application, various details of the embodiments of the present application are included to facilitate understanding, but these details should be construed as merely exemplary. Therefore, those skilled in the art will 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. In addition, in the following description, for the sake of clarity and conciseness, descriptions of well-known functions and structures will be omitted.

[0042] Terms and Definitions As used herein, the term "alkyl" refers to an optionally substituted group containing one or more carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more carbon atoms). The term "C1-C14 alkyl" refers to an optionally substituted, straight or branched chain saturated hydrocarbon containing 1 to 14 carbon atoms. Unless otherwise indicated, alkyl groups described herein refer to both unsubstituted and substituted alkyl groups.

[0043] As used herein, the term "carbocycle" or "carbocyclyl" refers to an optionally substituted monocyclic or polycyclic ring system containing one or more carbon rings. The ring can be 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13-, 14-, 15-, 16-, 17-, 18-, 19-, or 20-membered. The term "5- to 7-membered ring" refers to a monocyclic, fused, or spirocarbocycle having 5 to 7 carbon atoms. A carbocycle may contain one or more carbon-carbon double or triple bonds and may be non-aromatic or aromatic (e.g., cycloalkyl or aryl). Examples of carbocycles include cyclopentyl, cyclohexyl, phenyl, naphthyl, and 1,2-dihydronaphthyl.

[0044] As used herein, "cycloalkyl" refers to a non-aromatic carbocyclic ring, which may or may not contain double or triple bonds. Unless otherwise indicated, carbocyclic rings described herein refer to unsubstituted and substituted carbocyclic groups, i.e., optionally substituted carbocyclic rings.

[0045] As used herein, the term "heterocycle" or "heterocyclyl" refers to an optionally substituted monocyclic or polycyclic ring system containing one or more rings, at least one of which contains at least one heteroatom (e.g., 1, 2, 3, 4, or 5 heteroatoms, which may be, for example, nitrogen, oxygen, or sulfur atoms). The ring can be 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered. The heterocycle can contain one or more double or triple bonds and can be non-aromatic or aromatic (e.g., heterocycloalkyl or heteroaryl). Examples of heterocycles include imidazolyl, imidazolinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyryl, pyrrolyl group, pyrrolyl, pyrrolyl, furanyl, thienyl, pyrrolyl, pyrrolyl, pyrrolyl, furyl, thienyl, and phenyl.

[0046] As used herein, the term "heterocycloalkyl" refers to a non-aromatic heterocycle, which may or may not contain double or triple bonds. Unless otherwise indicated, heterocycle as used herein refers to unsubstituted and substituted heterocyclic groups, i.e., optionally substituted heterocycles.

[0047] As used herein, "aryl" refers to an optionally substituted carbocyclic group containing one or more aromatic rings. Examples of aryl groups include phenyl and naphthyl.

[0048] As used herein, "heteroaryl" refers to an optionally substituted heterocyclic group containing one or more aromatic rings. Examples of heteroaryl include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, and thiazolyl. Both aryl and heteroaryl can be optionally substituted. Unless otherwise specified, aryl or heteroaryl as used herein refers to both unsubstituted and substituted groups, i.e., optionally substituted aryl or heteroaryl.

[0049] Unless otherwise specified, alkyl groups can be optionally substituted. Optional substituents include halogen atoms (e.g., chloro, bromo, fluoro, or iodo), carboxylic acids (e.g., —C(O)OH), alcohols (e.g., hydroxyl, —OH), esters (e.g., —C(O)OR or —OC(O)R), aldehydes (e.g., —C(O)H), carbonyl groups (e.g., represented by —C(O)R or C═O), acyl halides (e.g., —C(O)X, [wherein , X is a halide selected from bromide, fluoride, chloride, and iodide], carbonate (e.g., —OC(O)OR), alkoxy (e.g., —OR), acetal, phosphate, thiol (e.g., —SH), sulfoxide (e.g., —S(O)R), sulfite group (e.g., —S(O)OH), sulfonate group (e.g., —S(O)OH), thiol (e.g., —C(S)H), sulfate In some embodiments, the substituent may be selected from, but not limited to, alkyl, sulfonyl (e.g., -S(O)-), amido (e.g., -C(O)NR or -N(R)C(O)R), azido (e.g., -N), nitro (e.g., -NO), cyano (e.g., -CN), isocyano (e.g., -NC), acyloxy (e.g., -OC(O)R), amino (e.g., -NR, -NRH, or -NH), carbamoyl (e.g., -OC(O)NR, -OC(O)NRH, or -OC(O)NH), sulfonamido, alkyl, alkenyl, and cyclic groups (e.g., carbocyclyl or heterocyclyl). In any one of the foregoing, R is alkyl or alkenyl as defined herein. In some embodiments, the substituent itself may be further substituted, for example, with 1, 2, 3, 4, 5, or 6 substituents as defined herein. For example, C 1~6 Alkyl can be further substituted with 1, 2, 3, 4, 5, or 6 substituents described herein.

[0050] As used herein, the term "compound" is meant to include all isomers and isotopes of the depicted structure. "Isotopes" refer to atoms with the same atomic number but different mass numbers due to the different number of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium. Furthermore, the compounds, salts, or complexes of the present application can be used to form solvates and hydrates by combining with solvents or water molecules and preparing them by conventional methods.

[0051] As used herein, the term "contact" refers to establishing a physical connection between two or more entities. For example, contacting a mammalian cell with a nanoparticle composition means that the mammalian cell and the nanoparticle share a physical connection. Methods for contacting cells with external entities both in vivo and ex vivo are well known in the field of biology. For example, a nanoparticle composition can contact mammalian cells in a mammal by various routes of administration (e.g., intravenous, intramuscular, intradermal, and subcutaneous), and various amounts of the nanoparticle composition can be involved. In addition, a nanoparticle composition can contact two or more mammalian cells.

[0052] As used herein, the term "delivery" refers to providing an entity to a target site. For example, delivering a therapeutic and / or prophylactic agent to a subject can include administering to the subject (e.g., by intravenous, intramuscular, intradermal, or subcutaneous routes) a nanoparticle composition containing the therapeutic and / or prophylactic agent. Administering a nanoparticle composition to a mammal or mammalian cells can include contacting one or more cells with the nanoparticle composition.

[0053] As used herein, the term "enhancing delivery" refers to the delivery of a greater (e.g., at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold) amount of therapeutic and / or prophylactic agent by a nanoparticle to a target tissue of interest (e.g., the liver of a mammal) compared to the level of therapeutic and / or prophylactic agent delivered to the target tissue (e.g., MC3, KC2, or DLinDMA) by a control nanoparticle. The comparison can be made by comparing the amount of protein produced in the tissue to the weight of the tissue, comparing the amount of therapeutic and / or prophylactic agent in the tissue to the weight of the tissue, comparing the amount of protein produced in the tissue to the total amount of protein in the tissue, or comparing the amount of therapeutic and / or prophylactic agent in the tissue to the total amount of therapeutic and / or prophylactic agent in the tissue. It should be understood that enhanced delivery of nanoparticles to a target tissue need not be determined in the subject being treated but can be determined in alternatives, such as an animal model (e.g., a rat model). In certain embodiments, nanoparticle compositions comprising a compound of Formula (I), (IA), (IB), (IC), (ID) have substantially the same level of delivery enhancement regardless of the route of administration.

[0054] As used herein, the terms "specific delivery" or "specifically delivering" refer to the delivery of a greater (e.g., at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold) amount of therapeutic and / or prophylactic agent by a nanoparticle to a target tissue (e.g., the liver, spleen, stomach, intestinal tract of a mammal) compared to a non-target tissue. The level of nanoparticle delivery to a particular tissue can be measured by comparing the weight of protein produced in the tissue to the weight of the tissue, comparing the amount of therapeutic and / or prophylactic agent in the tissue to the weight of the tissue, comparing the weight of protein produced in the tissue to the weight of total protein in the tissue, or comparing the amount of therapeutic and / or prophylactic agent in the tissue to the total amount of therapeutic and / or prophylactic agent in the tissue.

[0055] As used herein, "encapsulation efficiency" refers to the amount of therapeutic and / or prophylactic agent that becomes part of the nanoparticle composition relative to the total amount of therapeutic and / or prophylactic agent used in preparing the nanoparticle composition. For example, if 97 mg of therapeutic and / or prophylactic agent are encapsulated in the nanoparticle composition out of a total of 100 mg of therapeutic and / or prophylactic agent initially provided to the composition, the encapsulation efficiency may be 97%. As used herein, "encapsulation" may refer to complete, substantial, or partial encapsulation, closure, enclosing, or enveloping.

[0056] As used herein, "expression" of a nucleic acid sequence refers to translation of mRNA into a polypeptide or protein and / or post-translational modification of a polypeptide or protein.

[0057] As used herein, the term "in vitro" refers to events that do not occur within a living organism (e.g., an animal, plant, or microorganism), but rather to events that occur in an artificial environment, such as in a test tube or reaction vessel, in a cell culture, in a culture dish, etc.

[0058] As used herein, the term "in vivo" refers to events that take place within an organism (e.g., an animal, plant, or microorganism, or cells or tissues thereof).

[0059] As used herein, the term "ex vivo" refers to an event that occurs outside of an organism (e.g., an animal, plant, or microorganism, or cells or tissues thereof). An ex vivo event may occur in an environment that is minimally altered from the natural (e.g., in vivo) environment.

[0060] As used herein, the term "isomer" refers to a geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer, or diastereoisomer of a compound. A compound 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 for resolving them into their component enantiomers or stereoisomers are well known.

[0061] As used herein, a "lipid component" is a component of a nanoparticle composition that includes one or more lipids. For example, the lipid component can include one or more cationic / pH-responsive lipids, PEGylated lipids, structured lipids, or other lipids such as phospholipids.

[0062] As used herein, a "linker" refers to a moiety that connects two moieties, such as the linkage between two nucleosides of a cap. The linker may contain one or more groups, including, but not limited to, a phosphate group (e.g., phosphate, borophosphate, thiophosphate, selenophosphate, and phosphonate), alkyl, amide, or glycerol. For example, the two nucleosides of a cap analog may be linked at their 5' positions by a triphosphate group or by a chain containing two phosphate moieties and a borophosphate moiety.

[0063] As used herein, "administration method" can include intravenous administration, intramuscular administration, intradermal administration, subcutaneous administration, or other methods of delivering a composition to a subject. Any one of the administration methods can be selected for targeted delivery (e.g., specific delivery) to a particular area or system of the body.

[0064] As used herein, "modified" refers to something that does not occur in nature. For example, RNA can be modified RNA. That is, RNA can contain one or more non-naturally occurring nucleobases, nucleosides, nucleotides, or linkers. "Modified" materials are also referred to herein as "engineered" materials. Substances can be chemically, structurally, or functionally modified or altered. For example, modified nucleobase species can contain one or more non-naturally occurring substitutions.

[0065] As used herein, a "nanoparticle composition" refers to a composition containing one or more lipids. The particle size of a nanoparticle composition is typically on the order of a few microns or less and may contain a lipid bilayer. Nanoparticle compositions include lipid nanoparticles (LNPs), liposomes (e.g., lipid vesicles), and lipid complexes. For example, a nanoparticle composition may be a liposome having a lipid bilayer with a diameter of 500 nm or less.

[0066] As used herein, "naturally occurring" means existing in nature without artificial assistance.

[0067] As used herein, "patient" refers to a subject who may seek or need treatment, who needs treatment, who is receiving treatment, who is about to receive treatment, or who is receiving care for a particular disorder by a trained professional.

[0068] As used herein, "PEG lipid" or "PEGylated lipid" refers to a lipid that includes a polyethylene glycol moiety.

[0069] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0070] As used herein, the phrase "pharmaceutically acceptable excipient" refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending, complexing, or dissolving an active compound), which is substantially non-toxic and non-inflammatory to a patient. Excipients can include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), wetting agents, emulsifiers, fillers (diluents), film-forming or coating agents, flavors, fragrances, glidants (flow improvers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water for hydration. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E (α-tocopherol), vitamin C, xylitol, and others disclosed herein.

[0071] In this application, the structural formula of a compound shows a specific isomer for convenience, but it should be understood that this application includes all isomers, such as geometric isomers, optical isomers based on asymmetric carbons, stereoisomers, tautomers, etc., and not all isomers may have the same activity level. In addition, the compound represented by the structural formula of the compound described in this application may exist as a crystalline polymorph. It should be noted that any crystalline form, a mixture of crystalline forms, or an anhydrate or hydrate thereof is included within the scope of this application.

[0072] The terms "crystalline polymorph," "polymorph," or "crystalline form" refer to a crystalline structure in which a compound (or a salt or solvate thereof) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability, and solubility. Recrystallization solvent, crystallization rate, storage temperature, and other factors may cause one form to predominate. Crystalline polymorphs of a compound can be prepared by crystallization under different conditions.

[0073] The nanoparticle compositions of the present application may further include one or more salts of the compounds. The salts may be pharmaceutically acceptable salts. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety into its salt form (e.g., by reacting the free base with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; base or organic salts of acidic residues such as carboxylic acids. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxyethanesulfonate, lactate, lauric acid, and the like. Examples of the salts include phosphate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, hexadecanoate, pectinate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, tosylate, undecanoate, and valerate.

[0074] Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., and non-toxic ammonium, quaternary ammonium, and amine cations include, but are not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Pharmaceutically acceptable salts of the present application include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Typically, these salts can be prepared by reacting the free acidic or basic form of these compounds with a chemically calculated amount of the appropriate base or acid in water or an organic solvent, or a mixture of both. Typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

[0075] As used herein, a "phospholipid" is a lipid containing a phosphate moiety and one or more carbon chains (e.g., unsaturated fatty acid chains). A phospholipid may contain one or more multiple (e.g., double or triple) bonds (e.g., one or more unsaturated bonds). Certain phospholipids may promote fusion with membranes. For example, cationic phospholipids may interact with one or more negatively charged phospholipids in a membrane (e.g., a cell membrane or intracellular membrane). The fusion of a phospholipid with a membrane allows one or more components of a lipid-containing composition to pass through the membrane, thereby, for example, delivering one or more components to a cell.

[0076] As used herein, "polydispersity index" is a ratio that describes the uniformity of the particle size distribution of a system. A relatively small value, e.g., less than 0.3, indicates a narrow particle size distribution.

[0077] As used herein, the term "polypeptide" or "polypeptide of interest" refers to a polymer of amino acid residues, typically joined by peptide bonds, which may be naturally occurring (e.g., isolated or purified) or synthetically produced.

[0078] As used herein, "RNA" refers to a ribonucleic acid, which may or may not occur naturally. For example, the RNA may contain modified and / or non-naturally occurring components, such as one or more nucleobases, nucleosides, nucleotides, or linkers. The RNA may contain a cap structure, a chain-terminating nucleoside, a stem-loop, a polyA sequence, and / or a polyadenylation signal. The RNA may have a nucleotide sequence encoding a polypeptide of interest. For example, the RNA may be messenger RNA (mRNA). Translation of an mRNA encoding a specific polypeptide (e.g., in vivo translation of the mRNA in a mammalian cell) may result in the encoded polypeptide. The RNA may be selected from the non-limiting group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (airRNA), microRNA (miRNA), double-stranded RNA (dsRNA), short hairpin RNA (shRNA), mRNA, and mixtures thereof.

[0079] As used herein, a "single unit dose" is a dose of any therapeutic agent administered in one dose / one time / single route / single point of contact, i.e., a single administration.

[0080] As used herein, "split dose" refers to a single unit dose or a dose in which the total daily dose is divided into two or more doses.

[0081] As used herein, a "total daily dose" is the amount administered or prescribed over a 24-hour period. This may be administered in a single unit dose.

[0082] As used herein, "particle size" or "average particle size" in the context of a nanoparticle composition refers to the average diameter of the nanoparticle composition.

[0083] As used herein, the term "subject" or "patient" refers to any living organism to which a composition according to the present application may be administered, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and / or plants.

[0084] As used herein, "target cell" refers to one or more cells of interest. The cells may be found in vitro, in vivo, in situ, or in the tissue or organ of an organism. The organism may be an animal, preferably a mammal, more preferably a human, and most preferably a patient.

[0085] As used herein, "target tissue" refers to one or more tissue types of interest where therapeutic and / or prophylactic delivery results in a desired biological and / or pharmacological effect. Examples of target tissues include specific tissues, organs, and systems or groups thereof. In particular applications, the target tissue may be the kidney (e.g., intracoronary or intrafemoral), or the kidney, lung, spleen, or vascular endothelium (e.g., by intratumoral injection). "Non-target tissue" refers to any tissue type where expression of the encoded protein does not result in a desired biological and / or pharmacological effect. In particular applications, non-target tissues may include the liver and spleen.

[0086] The terms "therapeutic agent" or "prophylactic agent" refer to any agent that has a therapeutic, diagnostic, and / or prophylactic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject. Therapeutic agents are also called "active agents" or "active ingredients." Such substances include, but are not limited to, cytotoxins, radioactive ions, chemotherapeutic agents, small molecule drugs, proteins, and nucleic acids.

[0087] As used herein, the term "therapeutically effective amount" refers to a sufficient quantity of an agent (e.g., nucleic acid, drug, composition, therapeutic agent, diagnostic agent, prophylactic agent, etc.) to be delivered for administration to a subject suffering from or susceptible to an infection, disease, disorder, and / or condition for the purposes of treating, ameliorating, diagnosing, preventing the symptoms of, and / or delaying the onset of, the infection, disease, disorder, and / or condition.

[0088] As used herein, "transfection" refers to the introduction of a species (e.g., RNA) into a cell. Transfection can occur, for example, in vitro, ex vivo, or in vivo.

[0089] As used herein, the term "treatment" refers to the partial or complete response, alleviation, improvement, remission, delay in onset, inhibition of progression, reduction in severity, and / or reduction in the incidence of one or more symptoms or signs of a particular infection, disease, disorder, and / or condition. For example, "treating" cancer may refer to inhibiting tumor survival, growth, and / or spread. To reduce risk, subjects who do not exhibit the disease, disorder, and / or condition, a pathologically advanced condition associated with the disease, disorder, and / or condition, and / or who exhibit only early signs of the disease, disorder, and / or condition, a pathologically advanced condition associated with the disease, disorder, and / or condition, can be treated.

[0090] The present application discloses a compound according to formula (I), or a salt or isomer thereof:

[0091] [ka] wherein X and Y are independently selected from O or S; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; R1 is

[0092] [ka] (In the formula, R 1a and R 1b are independently selected from H or C1-C6 alkyl), and R2, R3 are independently selected from H or C1-C6 alkyl, or R1 and R2 together with the carbon atoms to which they are attached form a substituted or unsubstituted 5- to 7-membered nitrogen-containing heterocycle, and R3 is selected from H or C1-C6 alkyl, or R1 and R3 together with the carbon atoms to which they are attached form a ring A which is a substituted or unsubstituted carbocyclic or heterocyclic ring, and R2 is selected from H or C1-C6 alkyl; R4, R5, and R6 are independently selected from C1-C14 alkyl; m and n are independently selected from 0, 1, and 2, with the proviso that m and n are not both 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; q is selected from 0, 1, 2, 3, 4, 5 or 6.

[0093] In a preferred embodiment, R1 is

[0094] [ka] and in a specific embodiment, the compound of formula (I) is as shown in formula (IA):

[0095] [ka] wherein q is preferably selected from 0, 1, 2, 3 or 4. More preferably, q is selected from 2 or 3. More preferably, q is selected from 2.

[0096] In a preferred embodiment, the compound of formula (I) is of formula (IA):

[0097] [ka] wherein q is selected from 2 or 3; R 1a and R 1b is selected from -CH3, -CH2CH3 or -CH(CH3)2; R2 is H and R3 is H; n is 1 and m is 0, X and Y are both O, o is 5 or 6, p is 7, X1 and X2 are both C=O and Y1 and Y2 are O, or X1 is O, Y1 is C=O, X2 is C=O and Y2 is O; R4 is a C10 straight chain alkyl or a C11 straight chain alkyl; R5 and R6 are C8 straight chain alkyls. More preferably, R 1a and R 1b are both selected from -CH3.

[0098] In a preferred embodiment, R1 and R2 together with the carbon atom to which they are attached form a substituted or unsubstituted 5- to 7-membered nitrogen-containing heterocycle, and R3 is selected from H or C1-C6 alkyl, and in a specific embodiment, the compound of formula (I) is as shown in formula (IB):

[0099] [ka] wherein R7 is selected from H or C1-C6 alkyl; and r is selected from 0, 1, 2, or 3.

[0100] In a preferred embodiment, R1 and R3, together with the carbon atoms to which they are attached, form a ring A, which is a substituted or unsubstituted 5- to 7-membered carbocyclic or heterocyclic ring; R2 is selected from H or C1-C6 alkyl; in a specific embodiment, the compound of formula (I) is as shown in formula (IC):

[0101] [ka] wherein ring A is substituted by one or more R8, and R8 is

[0102] [ka] (In the formula, R 8a and R 8b are independently selected from H or C1-C6 alkyl, and s is selected from 0, 1, 2, 3, 4, 5, or 6).

[0103] In a specific embodiment, the compound of formula (I) is as shown in formula (ID):

[0104] [ka] wherein ring A is substituted by one or more R8, and R8 is

[0105] [ka] (In the formula, R 8a and R 8b are independently selected from H or C1-C6 alkyl, and s is selected from 0, 1, 2, 3, 4, 5, or 6).

[0106] In a specific embodiment, the ring A is

[0107] [ka] and the compound of formula (IC) is (IC-1),

[0108] [ka] As shown in; The compound of formula (ID) is (ID-1),

[0109] [ka] As shown in; In a specific embodiment, the ring A is

[0110] [ka] and the compound of formula (IC) is (IC-2),

[0111] [ka] As shown in; The compound of formula (ID) is (ID-2),

[0112] [ka] As shown in; In a specific embodiment, the ring A is

[0113] [ka] and the compound of formula (IC) is (IC-3),

[0114] [ka] As shown in; The compound of formula (ID) is (ID-3),

[0115] [ka] As shown in; In a specific embodiment, the ring A is

[0116] [ka] and the compound of formula (IC) is (IC-3),

[0117] [ka] The compound of formula (IC) is as shown in (IC-4),

[0118] [ka] or the compound of formula (IC) is as shown in (IC-5),

[0119] [ka] As shown in; The compound of formula (ID) is (ID-3),

[0120] [ka] and the compound of formula (ID) is (ID-4),

[0121] [ka] or the compound of formula (ID) is as shown in (ID-5),

[0122] [ka] As shown in

[0123] In a preferred embodiment, in the above formulas (I), (IA), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (ID), (ID-1), (ID-2), (ID-3), (ID-4), and (ID-5), R4 is selected from C1 to C14 alkyl, for example, R4 is selected from C2 to C14 alkyl, R4 is selected from C3 to C14 alkyl, R4 is selected from C4 to C14 alkyl, R4 is selected from C5 to C14 alkyl, and R4 is selected from C6 to C14 alkyl; R4 is selected from C7 to C14 alkyl, R4 is selected from C8 to C14 alkyl, R4 is selected from C9 to C14 alkyl, R4 is selected from C10 to C14 alkyl, C10 to C13 alkyl, C10 to C12 alkyl, or C10 to C11 alkyl, for example, R4 is C1 alkyl, R4 is C2 alkyl, R4 is C3 alkyl, R4 is C4 alkyl, R4 is C5 alkyl, R4 is C6 alkyl, R4 is C7 alkyl, R4 is C8 alkyl, R4 is C9 alkyl, R4 is C10 alkyl, R4 is C11 alkyl, R4 is C12 alkyl, R4 is C13 alkyl, or R4 is C14 alkyl. In a more preferred embodiment, R4 is C10 alkyl; in a more preferred embodiment, R4 is C11 alkyl.

[0124] In a preferred embodiment, R4 is selected from C1 to C14 straight chain alkyl, for example, R4 is selected from C2 to C14 straight chain alkyl, R4 is selected from C3 to C14 straight chain alkyl, R4 is selected from C4 to C14 straight chain alkyl, R4 is selected from C5 to C14 straight chain alkyl, R4 is selected from C6 to C14 straight chain alkyl, R4 is selected from C7 to C14 straight chain alkyl, R4 is selected from C8 to C14 straight chain alkyl, R4 is selected from C9 to C14 straight chain alkyl, R4 is selected from C10 to C14 straight chain alkyl, C10 to C13 straight chain alkyl, C1 It is selected from C10 to C12 straight-chain alkyl or C10 to C11 straight-chain alkyl, for example, R4 is C1 alkyl, R4 is C2 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, or R4 is C14 straight-chain alkyl. In a more preferred embodiment, R4 is C10 straight-chain alkyl; in a more preferred embodiment, R4 is C11 straight-chain alkyl.

[0125] In a specific embodiment, the compound of formula (I) has the formula (IA-1):

[0126] [ka] As shown in

[0127] In a specific embodiment, the compound of formula (I) has the formula (IA-2):

[0128] [ka] As shown in

[0129] In a specific embodiment, the compound of formula (I) has the formula (IB-1):

[0130] [ka] As shown in

[0131] In a specific embodiment, the compound of formula (I) has the formula (IB-2):

[0132] [ka] As shown in

[0133] In a specific embodiment, the compound of formula (I) has the formula (IC-6):

[0134] [ka] As shown in

[0135] In a specific embodiment, the compound of formula (I) has the formula (IC-7):

[0136] [ka] As shown in

[0137] In a specific embodiment, the compound of formula (I) has the formula (ID-6):

[0138] [ka] As shown in

[0139] In a specific embodiment, the compound of formula (I) has the formula (ID-7):

[0140] [ka] As shown in

[0141] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), In (IC-7), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), and (ID-7), R5 is selected from C1 to C14 alkyl, for example, R5 is selected from C2 to C14 alkyl, R5 is selected from C3 to C14 alkyl, R5 is selected from C4 to C14 alkyl, and R5 is selected from C5 to C14 alkyl, R5 is R5 is selected from C6 to C14 alkyl, R5 is selected from C7 to C14 alkyl, R5 is selected from C8 to C14 alkyl, R5 is selected from C8 to C13 alkyl, R5 is selected from C8 to C12 alkyl, R5 is selected from C8 to C11 alkyl, R5 is selected from C8 to C10 alkyl, or R5 is selected from C8 to C9 alkyl, for example, R5 is C1 alkyl, R5 is C2 alkyl, R5 is C3 alkyl, R5 is C4 alkyl, R5 is C5 alkyl, R5 is C6 alkyl, R5 is C7 alkyl, R5 is C8 alkyl, R5 is C9 alkyl, R5 is C10 alkyl, R5 is C11 alkyl, R5 is C12 alkyl, R5 is C13 alkyl, or R5 is C14 alkyl. In a preferred embodiment, R5 is C8 alkyl.

[0142] In a preferred embodiment, R5 is selected from C1 to C14 straight chain alkyl, for example, R5 is selected from C2 to C14 straight chain alkyl, R5 is selected from C3 to C14 straight chain alkyl, R5 is selected from C4 to C14 straight chain alkyl, R5 is selected from C5 to C14 straight chain alkyl, R5 is selected from C6 to C14 straight chain alkyl, R5 is selected from C7 to C14 straight chain alkyl, R5 is selected from C8 to C14 straight chain alkyl, R5 is selected from C8 to C13 straight chain alkyl, R5 is selected from C8 to C12 straight chain alkyl, R5 is selected from C8 to C11 straight chain alkyl; R5 is selected from C8 to C10 straight-chain alkyl, or R5 is selected from C8 to C9 straight-chain alkyl, for example, R5 is C1 alkyl, R5 is C2 alkyl, R5 is C3 straight-chain alkyl, R5 is C4 straight-chain alkyl, R5 is C5 straight-chain alkyl, R5 is C6 straight-chain alkyl, R5 is C7 straight-chain alkyl, R5 is C8 straight-chain alkyl, R5 is C9 straight-chain alkyl, R5 is C10 straight-chain alkyl, R5 is C11 straight-chain alkyl, R5 is C12 straight-chain alkyl, R5 is C13 straight-chain alkyl, or R5 is C14 straight-chain alkyl. In a preferred embodiment, R5 is C8 straight-chain alkyl.

[0143] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), In (IC-7), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), and (ID-7), R6 is selected from C1 to C14 alkyl, for example, R6 is selected from C2 to C14 alkyl, R6 is selected from C3 to C14 alkyl, R6 is selected from C4 to C14 alkyl, and R6 is selected from C5 to C14 alkyl, R6 is R6 is selected from C6 to C14 alkyl, R6 is selected from C7 to C14 alkyl, R6 is selected from C8 to C14 alkyl, R6 is selected from C8 to C13 alkyl, R6 is selected from C8 to C12 alkyl, R6 is selected from C8 to C11 alkyl, R6 is selected from C8 to C10 alkyl, or R6 is selected from C8 to C9 alkyl, for example, R6 is C1 alkyl, R6 is C2 alkyl, R6 is C3 alkyl, R6 is C4 alkyl, R6 is C5 alkyl, R6 is C6 alkyl, R6 is C7 alkyl, R6 is C8 alkyl, R6 is C9 alkyl, R6 is C16 alkyl, R6 is C11 alkyl, R6 is C12 alkyl, R6 is C13 alkyl, or R6 is C14 alkyl. In a preferred embodiment, R6 is C8 alkyl.

[0144] In a preferred embodiment, R6 is selected from C1 to C14 straight chain alkyl, for example, R6 is selected from C2 to C14 straight chain alkyl, R6 is selected from C3 to C14 straight chain alkyl, R6 is selected from C4 to C14 straight chain alkyl, R6 is selected from C5 to C14 straight chain alkyl, R6 is selected from C6 to C14 straight chain alkyl, R6 is selected from C7 to C14 straight chain alkyl, R6 is selected from C8 to C14 straight chain alkyl, R6 is selected from C8 to C13 straight chain alkyl, R6 is selected from C8 to C12 straight chain alkyl, R6 is selected from C8 to C11 straight chain alkyl. and R6 is selected from C8 to C10 straight-chain alkyl, or R6 is selected from C8 to C9 straight-chain alkyl, for example, R6 is C1 alkyl, R6 is C2 alkyl, R6 is C3 straight-chain alkyl, R6 is C4 straight-chain alkyl, R6 is C5 straight-chain alkyl, R6 is C6 straight-chain alkyl, R6 is C7 straight-chain alkyl, R6 is C8 straight-chain alkyl, R6 is C9 straight-chain alkyl, R6 is C10 straight-chain alkyl, R6 is C11 straight-chain alkyl, R6 is C12 straight-chain alkyl, R6 is C13 straight-chain alkyl, or R6 is C14 straight-chain alkyl. In a preferred embodiment, R6 is a C8 straight-chain alkyl.

[0145] In a preferred embodiment, the R5 is C8 alkyl and the R6 is C8 alkyl.

[0146] In a preferred embodiment, R5 is a C8 straight chain alkyl and R6 is a C8 straight chain alkyl.

[0147] In a specific embodiment, the compound of formula (I) has the formula (IA-3):

[0148] [ka] As shown in

[0149] In a specific embodiment, the compound of formula (I) has the formula (IB-3):

[0150] [ka] As shown in

[0151] In a specific embodiment, the compound of formula (I) has the formula (IC-8):

[0152] [ka] As shown in

[0153] In a specific embodiment, the compound of formula (I) has the formula (ID-8):

[0154] [ka] As shown in

[0155] In a preferred embodiment, o is 5: In a specific embodiment, the compound of formula (I) has the formula (IA-4):

[0156] [ka] As shown in

[0157] In a specific embodiment, the compound of formula (I) has the formula (IB-4):

[0158] [ka] As shown in

[0159] In a specific embodiment, the compound of formula (I) has the formula (IC-9):

[0160] [ka] As shown in

[0161] In a specific embodiment, the compound of formula (I) has the formula (ID-9):

[0162] [ka] As shown in

[0163] In a preferred embodiment, o is 6: In a specific embodiment, the compound of formula (I) has the formula (IA-5):

[0164] [ka] As shown in

[0165] In a specific embodiment, the compound of formula (I) has the formula (IB-5):

[0166] [ka] As shown in

[0167] In a specific embodiment, the compound of formula (I) has the formula (IC-10):

[0168] [ka] As shown in

[0169] In a specific embodiment, the compound of formula (I) has the formula (ID-10):

[0170] [ka] As shown in

[0171] In a preferred embodiment, p is 7: In a specific embodiment, the compound of formula (I) has the formula (IA-6):

[0172] [ka] As shown in

[0173] In a specific embodiment, the compound of formula (I) has the formula (IB-6):

[0174] [ka] As shown in

[0175] In a specific embodiment, the compound of formula (I) has the formula (IC-11):

[0176] [ka] As shown in

[0177] In a specific embodiment, the compound of formula (I) has the formula (ID-11):

[0178] [ka] As shown in

[0179] In a preferred embodiment, the compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-1) ), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), X1 and X2 are C=O, and Y1 and Y2 are O.

[0180] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-1), (I In (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), X1 is O, Y1 is C=O, X2 is C=O, and Y2 is O.

[0181] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-1), (I In (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), X1 is C=O, Y1 is O, X2 is O, and Y2 is C=O.

[0182] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC In (ID-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), X1 and X2 are O, and Y1 and Y2 are C=O.

[0183] In a preferred embodiment, the compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (I In C), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), both X and Y are O.

[0184] In a preferred embodiment, the compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (I In C), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), n ​​is 0 and m is 1.

[0185] In a preferred embodiment, the compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (I In C), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), n ​​is 1 and m is 0.

[0186] In a preferred embodiment, the compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (I In C), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), n ​​is 1 and m is 1.

[0187] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (IC-12), (IC-13), (IC-14), (IC-15), (IC-16), (IC-17), (IC-18), (IC-19), (IC-20), (IC-21), (IC-22), (IC-23), (IC-24), (IC-25), (IC-26), (IC-27), (IC-28), (IC-29), (IC-29), (IC-21), (IC-22), (IC-23), (IC-24), (IC-25), (IC-26), (IC-27), (IC-28), (IC-29), (IC-29), (IC-29), (IC-29), (IC-29), (IC-29), (IC-30), (IC-31), (IC-32), (IC-33), (IC-34), (IC-35), (IC-36), (IC-37), (IC-38), (IC-39 ...9), (IC-39), (IC-39), (IC-39), ( In (ID-1), (ID-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), the o is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0188] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (IC-12), (IC-13), (IC-14), (IC-15), (IC-16), (IC-17), (IC-18), (IC-19), (IC-20), (IC-21), (IC-22), (IC-23), (IC-24), (IC-25), (IC-26), (IC-27), (IC-28), (IC-29), (IC-29), (IC-21), (IC-22), (IC-23), (IC-24), (IC-25), (IC-26), (IC-27), (IC-28), (IC-29), (IC-29), (IC-29), (IC-29), (IC-29), (IC-29), (IC-30), (IC-31), (IC-32), (IC-33), (IC-34), (IC-35), (IC-36), (IC-37), (IC-38), (IC-39 ...9), (IC-39), (IC-39), (IC-39), ( In (ID-1), (ID-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0189] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), In (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), q is 0, 1, 2, 3, 4, 5, or 6.

[0190] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), In (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), o is 5 or 6.

[0191] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), In (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), p is 7 or 8.

[0192] In a preferred embodiment, the compounds of the above formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), In (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11), q is 2 or 3.

[0193] In a preferred embodiment, the compound of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (I B-6), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), 2 is H.

[0194] In a preferred embodiment, the compound of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (I B-6), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), 3 is H.

[0195] In a preferred embodiment, the compound of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (I B-6), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), 1a and R 1b is -CH3.

[0196] In a preferred embodiment, the compound of formula (I) is compound 1

[0197] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 2

[0198] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 3

[0199] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 4

[0200] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 5

[0201] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 6

[0202] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 7

[0203] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 8

[0204] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 9

[0205] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 10

[0206] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 11

[0207] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 12

[0208] [ka] is; In a preferred embodiment, the compound of formula (I) is compound 13

[0209] [ka] is.

[0210] The present application also relates to optical isomers of the following compounds:

[0211] [Table 1-1] [Table 1-2]

[0212] The present application further provides a method for compound (IA), comprising:

[0213] [ka] wherein Z1 is selected from OH, Z2 and Z3 are each independently selected from OH or SH, and Z4 is selected from OMs; X and Y are independently selected from O or S; X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; R 1a , R 1b are independently selected from H or C1-C6 alkyl; R2, R3 are independently selected from H or C1-C6 alkyl; R4, R5, and R6 are independently selected from C1-C14 alkyl; m and n are independently selected from 0, 1, or 2, with the proviso that m and n are not both 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; q is selected from 0, 1, 2, 3, 4, 5 or 6; The compound J reacts with the compound K in a benzene-based solution of pyridinium 4-methylbenzenesulfonate (Tso-Py) to form the compound L, and the benzene-based solvent used in the benzene-based solution can be toluene, xylene, trimethylbenzene, tetramethylbenzene, etc., and in a preferred embodiment of the present application, the benzene-based solvent is toluene; The compound L is reacted in a halogenated alkane solution of (CHSO)O to provide the compound M, where the halogenated alkane is abbreviated as haloalkane or alkyl halide, and refers to an organic compound in which one or more hydrogen atoms in an alkane molecule are replaced by halogen atoms (fluorine, chlorine, bromine, iodine). The haloalkane in the present application is not limited, and in a preferred embodiment, the chlorinated alkane is preferably DCM, dichloroethane, or chloroform; Compound M reacts in a polar aprotic solvent to form compound IA, which can be dimethyl sulfoxide (DMSO), acetone, acetonitrile, dimethylformamide, dimethylacetamide, hexamethylphosphoramide, tetrahydrofuran (THF), N,N-dimethylformamide (DMF), dimethylacetamide (DMA), preferably tetrahydrofuran (THF).

[0214] In the above synthesis method, the source of compound J is not limited, and may be commercially available or internally synthesized. In a specific embodiment, compound J is synthesized by the following synthesis method.

[0215] [ka] wherein X1 and X2 are independently selected from C=O or O, and Y1 and Y2 are independently selected from C=O or O, with the proviso that X1 and Y1 are not both C=O or O, and X2 and Y2 are not both C=O or O; X is selected from halogens; R4, R5, and R6 are independently selected from C1-C14 alkyl; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; Compound H reacts with compound C in the presence of a carbonate, NBuI, and a polar aprotic solvent to produce compound I (Ts represents p-toluenesulfonyl); in a preferred embodiment, the carbonate is an alkali metal carbonate, preferably CsCO or KCO; The polar aprotic solvent may be dimethyl sulfoxide (DMSO), acetone, acetonitrile, dimethylformamide, dimethylacetamide, hexamethylphosphoramide, tetrahydrofuran (THF), N,N-dimethylformamide (DMF), dimethylacetamide (DMA), preferably DMF or DMA; The compound I reacts at pH 1-5 to give the compound J, preferably the pH is adjusted by hydrochloric acid solution, the concentration of hydrochloric acid is preferably 12M, and the reaction solvent is a haloalkane, preferably DCM, dichloroethane or chloroform.

[0216] In the above synthesis method, the source of compound H is not limited, and it may be commercially available or internally synthesized. In a specific embodiment, compound H is synthesized by the following synthesis method.

[0217] [ka] wherein X1 and Y1 are independently selected from C=O or O; X is selected from halogens; R4 is selected from C1-C14 alkyl; o is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; The compound F is reacted with the compound G in the presence of a carbonate, NBuI, and a polar aprotic solvent to provide the compound H; in a preferred embodiment, the carbonate is an alkali metal carbonate, preferably CsCO or KCO; The polar aprotic solvent may be dimethyl sulfoxide (DMSO), acetone, acetonitrile, dimethylformamide, dimethylacetamide, hexamethylphosphoramide, tetrahydrofuran (THF), N,N-dimethylformamide (DMF), dimethylacetamide (DMA), preferably DMF or DMA.

[0218] The present application relates to compounds of formula (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), Further provided are nanoparticle compositions comprising a lipid component, the nanoparticle compositions comprising the compounds (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), and (ID-11).

[0219] In some embodiments, the nanoparticle composition has a diameter of 50 nm to 110 nm, e.g., 51 nm, 52 nm, 53 nm, 54 nm, 55 nm, 56 nm, 57 nm, 58 nm, 59 nm, 60 nm, 61 nm, 62 nm, 63 nm, 64 nm, 65 nm, 66 nm, 67 nm, 68 nm, 69 nm, 70 nm, 71 nm, 72 nm, 73 nm, 74 nm, 75 nm, 76 nm, 77 nm, 78 nm, 79 nm, 80 nm, 81 nm, 82 nm, 83 nm, 84 nm, 85 nm, 86 nm, 87 nm, 88 nm, 89 nm, 90 nm, 91 nm, 92 nm, 93 nm, 94 nm, 95 nm, 96 nm, 97 nm, 98 nm, 99 nm, 100 nm, 101 nm, 102 nm, 103 nm, 104 nm, 105 nm, 106 nm, 107 nm, 108 nm, 109 nm, 110 nm, 111 nm, 112 nm, 113 nm, 114 nm, 115 nm, 116 nm, 117 nm, 118 nm, 119 nm, 120 nm, 121 nm, 122 nm, 123 nm, 124 nm, 125 nm, 126 nm, 127 nm, 128 nm, 129 nm, 130 nm, 131 nm, 132 nm, 133 nm, 134 nm, 135 nm, 136 nm, 137 nm, 138 nm, 13 nm, 79 nm, 80 nm, 81 nm, 82 nm, 83 nm, 84 nm, 85 nm, 86 nm, 87 nm, 88 nm, 89 nm, 90 nm, 91 nm, 92 nm, 93 nm, 94 nm, 95 nm, 96 nm, 97 nm, 98 nm, 99 nm, 100 nm, 101 nm, 102 nm, 103 nm, 104 nm, 105 nm, 106 nm, 107 nm, 108 nm or 109 nm.

[0220] The nanoparticle composition can include, for example, lipid nanoparticles (LNPs), liposomes, lipid vesicles, and lipid complexes. In some embodiments, the nanoparticle composition is a vesicle comprising one or more lipid bilayers. In some embodiments, the nanoparticle composition comprises two or more concentric bilayers separated by two or more aqueous compartments. The lipid bilayers can be functionalized and / or crosslinked to each other. The lipid bilayers can comprise one or more ligands, proteins, or channels.

[0221] The nanoparticle compositions of the present application include a lipid component comprising at least one compound according to Formula (I), (IA), (IIA), (IB), (IC), or (ID). For example, the lipid component of the nanoparticle composition can include one or more of Compounds 1-32. The nanoparticle composition can also include various other components. For example, in addition to a compound according to Formula (I), (IA), (IIA), (IB), (IC), or (ID), the lipid component of the nanoparticle composition can also include one or more other lipids.

[0222] The lipid component of the nanoparticle composition may include one or more PEG-lipids or PEG-modified lipids. Alternatively, such substances may be referred to as PEGylated lipids. PEG-lipids are lipids modified with polyethylene glycol. The PEG-lipid 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.

[0223] The lipid component of the nanoparticle composition may comprise one or more structured lipids. The structured lipids may be selected from, but are not limited to, cholesterol, coprosterol, sitosterol, ergosterol, stigmasterol, and mixtures thereof. In some embodiments, the structured lipid is cholesterol. In some embodiments, the structured lipid comprises cholesterol and a corticosteroid (e.g., prednisolone, dexamethasone, prednisone, and hydrocortisone), or a combination thereof.

[0224] The lipid component of the nanoparticle composition may comprise one or more phospholipids, and the phospholipids used in the nanoparticle compositions and methods may be: Dilauryl lecithin (DLPC), Dimyristoylphosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dioleoylphosphatidylcholine (DUPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 lyso-PC), 1,2-divinyl-sn-glycero-3-phosphocholine, 1,2-diarylacyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-diethenol-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexaenoate-sn-glycero-3-phosphoethanolamine, It may be selected from 1,2-dioleoyl-sn-glycero-3-phosphate-(1-glycerol) sodium salt (DOPG) or sphingomyelin.

[0225] 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.

[0226] In some embodiments, nanoparticle compositions comprising one or more lipids described herein may further comprise one or more adjuvants, such as glucopyranosyl lipid adjuvant (GLA), CpG oligodeoxynucleotide (e.g., class A or class B), poly(I:C), aluminum hydroxide, and Pam3CSK4.

[0227] The nanoparticle composition can comprise one or more therapeutic and / or prophylactic agents.

[0228] 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, comprising administering to the mammal and / or contacting mammalian cells with a composition comprising therapeutic and / or prophylactic nanoparticles.

[0229] Therapeutic and / or prophylactic agents include biologically active substances and may alternatively be referred to as "active agents." Therapeutic and / or prophylactic agents may be substances that, when delivered to a cell or organ, effect a desired change in the cell, organ, or other body tissue or system. Such substances may be used to treat one or more diseases, disorders, or conditions. In some embodiments, therapeutic and / or prophylactic agents are small molecule drugs that may be used to treat specific diseases, disorders, or conditions. Examples of drugs that can be used in the nanoparticle compositions include antitumor agents (e.g., vincristine, adriamycin, mitoxantrone, camptothecin, cisplatin, bleomycin, cyclophosphamide, methotrexate, and streptozotocin), anti-tumor agents (e.g., actinomycin D, vincristine), vinblastine, cystine arabinoside, anthracyclines, alkylating agents, platinum compounds, antimetabolites and nucleoside analogs (e.g., methotrexate, purine, and pyrimidine analogs), anti-infectives, local anesthetics (e.g., dibucaine and chlorpromazine), beta-adrenergic blockers (e.g., propranolol, timolol, and labetalol), antihypertensives (e.g., benzodiazepines, benzocaine ... Examples of antihistamines include, but are not limited to, anti-inflammatory drugs (e.g., clonidine and hydralazine), antidepressants (e.g., imipramine, amitriptyline, and doxepin), antiepileptic drugs (e.g., phenytoin), antihistamines (e.g., diphenhydramine, chlorphenamine, and promethazine), antibiotics / antibacterial agents (e.g., gentamicin, ciprofloxacin, and cefoxitin), antifungal agents (e.g., miconazole, terconazole, econazole, isoconazole, butoconazole, clotrimazole, itraconazole, nystatin, and naftifine), anthelmintics, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, antiglaucoma agents, vitamins, anesthetics, and contrast media.

[0230] In some embodiments, the therapeutic and / or prophylactic agent is a cytotoxin, a radioactive ion, a chemotherapeutic agent, a vaccine, a compound that elicits an immune response, and / or another therapeutic and / or prophylactic agent. Cytotoxins or cytotoxic agents include any agent that may be harmful to cells. Examples include, but are not limited to, paclitaxel, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, adriamycin, daunorubicin, dihydroxyanthraquinone, cinmetasone, 1-nortestosterone, Aspergillus oryzae, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids, such as maytansinol, rachelmycin (CC-1065), and analogs or homologs thereof. Radioactive ions include, but are not limited to, iodine (e.g., iodine-125 or iodine-131), strontium-89, phosphorus, palladium, cesium, iridium, phosphate, cobalt, yttrium-90, and SA-153. Vaccines include compounds and preparations that can provide immunity against one or more conditions associated with infectious diseases (e.g., influenza, measles, human papillomavirus (HPV), rabies, meningitis, whooping cough, tetanus, plague, hepatitis, and tuberculosis), and include mRNA encoding antigens and / or epitopes from infectious diseases. Vaccines also include compounds and preparations that induce an immune response against cancer cells, and may include mRNA encoding tumor cell-derived antigens, epitopes, and / or neoepitopes. Compounds that induce an immune response may include vaccines, corticosteroids (e.g., dexamethasone), and other species.

[0231] Other therapeutic and / or prophylactic agents include antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarboxylase), alkylating agents (e.g., methyclothiazide, thiotepa-chloramphenicol, rachelmycin (CC-1065), melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomatase, thiazol-2-one ... Anti-inflammatory drugs include, but are not limited to, cis-dichlorodiamineplatinum(II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and antimitotic agents (e.g., vincristine, vinblastine, paclitaxel, and zeatin glycosides).

[0232] In some embodiments, the vaccine and / or compound capable of eliciting an immune response is selected from the group consisting of: (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), , (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), for example, a composition containing one or more of compounds 1 to 32 is administered intramuscularly. Other therapeutic and / or preventative measures include antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, dacarbazine), alkylating agents (e.g., methoxyethylamine, metsulfovax, chlorobutyric acid, rachelmycin (CC-1065), melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mycobacterial steroids, methicillin-resistant Staphylococcus aureus (MSA), ... Anti-inflammatory drugs include, but are not limited to, cis-dichlorodiamineplatinum(II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and adriamycin), antibiotics (e.g., actinomycin, mithramycin, adriamycin, actinomycin), anthramycin (AMC)), and antimitotic agents (e.g., vincristine, vinblastine, paclitaxel, and maytansinoids).

[0233] In other embodiments, the therapeutic and / or prophylactic agent is a protein. Therapeutic proteins that can be used in the nanoparticle compositions described herein include, but are not limited to, gentamicin, amikacin, insulin, erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), VIR factors, luteinizing hormone-releasing hormone (LHRH) analogs, interferon, eparin, hepatitis B surface antigen, typhoid vaccine, and cholera vaccine.

[0234] In some embodiments, the therapeutic agent is a polynucleotide or nucleic acid (e.g., ribonucleic acid or deoxyribonucleic acid). The term "polynucleotide," in its broadest sense, includes any compound and / or substance that is or may be attached to an oligonucleotide chain. Exemplary polynucleotides for use in accordance with the present invention include, but are not limited to, one or more of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), including messenger mRNA (mRNA), hybrids thereof, RNAi-inducing agents, RNase inhibitors, siRNA, shRNA, miRNA, antisense RNA, ribozymes, catalytic DNA, RNA that induces triple helix formation, aptamers, vectors, and the like. In some embodiments, the therapeutic and / or prophylactic agent is RNA. The RNA that can be used in the compositions and methods described herein can be selected from, but is not limited to, shortmers, antigametophytic, antisense, ribozymes, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), double-stranded RNA (dsRNA), short hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and mixtures thereof. In certain embodiments, the RNA is mRNA.

[0235] In certain embodiments, the therapeutic and / or prophylactic agent is an mRNA. The mRNA can encode any polypeptide of interest, including naturally occurring, non-naturally occurring, or modified polypeptides. The polypeptide encoded by the mRNA can be of any size and have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA can exert a therapeutic effect when expressed in a cell.

[0236] In other embodiments, the therapeutic and / or prophylactic agent is an siRNA. The siRNA can selectively knock down or downregulate the expression of a gene of interest. For example, the siRNA can be selected to silence a gene associated with a particular disease, disorder, or condition when a nanoparticle composition containing the siRNA is administered to a subject in need thereof. The siRNA can comprise a sequence complementary to an mRNA sequence encoding the gene or protein of interest. In some embodiments, the siRNA can be an immunomodulatory siRNA.

[0237] In some embodiments, the therapeutic and / or prophylactic agent is shRNA, or a vector or plasmid encoding shRNA.When an appropriate construct is delivered to the nucleus, shRNA can be produced in target cells.The constructs and mechanisms related to shRNA are well known in the relevant technical field.

[0238] The nanoparticle compositions of the present application can include one or more ingredients in addition to those described above. For example, the nanoparticle compositions can include one or more hydrophobic small molecules, such as vitamins (e.g., vitamin A or vitamin E) or sterols.

[0239] The nanoparticle compositions can also include one or more permeability enhancer molecules, carbohydrates, polymers, surface modifiers, or other components. The permeability enhancer molecules can be those described in U.S. Patent Application Publication No. 2005 / 0222064. The carbohydrates can include monosaccharides (e.g., glucose) and polysaccharides (e.g., glycogen and its derivatives and analogs).

[0240] The nanoparticle composition may further comprise one or more polymers, which may be biodegradable and / or biocompatible, selected from, but not limited to, polyamines, polyethers, polyamides, polyesters, polyurethanes, polyureas, polycarbonates, polystyrenes, polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyleneimines, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylesters. For example, polymers include polycaprolactone (PCL), ethylene vinyl acetate polymer (EVA), polylactic acid (PLA), poly-L-lactic acid (PLLA), polyglycolic acid (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide (PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-caprolactone-glycolide), poly(D,L-lactide-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), Polyalkyl cyanoacrylates, polyurethanes, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethylene glycol, poly-L-glutamic acid, polyhydroxy acids, polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene; polyalkylene glycols such as poly(ethylene glycol) (PEG); polyalkylene oxides (PEO); polyalkylene terephthalates such as polyethylene glycol terephthalate; polyvinyl alcohol (PVA);Polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as polyvinyl chloride (PVC), polyvinylpyrrolidone (PVP), polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitrocellulose, hydroxypropyl cellulose, carboxymethyl cellulose, acrylic polymers such as poly(methyl methacrylate) (PMMA), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and copolymers and mixtures thereof, polydioxanone and copolymers thereof, polyhydroxyalkanoates, polypropylene fumarate, polyoxymethylene, poloxamer, polyethylene diamine, poly(orthoesters), poly(butyric acid), poly(valeric acid), poly(lactide-caprolactone), trimethylene carbonate, poly(N-acryloylmorpholine) (PAcM), poly(2-methyl-2-oxazoline (PMOX), poly(2-ethyl-2-oxazoline) (PEOZ), and polyglycerol;

[0241] The nanoparticle composition may further comprise one or more surface modifiers, including, but not limited to, anionic proteins (e.g., bovine serum albumin), surfactants (cationic surfactants, e.g., dimethyloctacosylammonium bromide), sugars or sugar derivatives (e.g., cyclodextrin), nucleic acids, polymers (e.g., heparin, polyethylene glycol, and poloxamer), mucolytic agents (e.g., acetylcysteine, artemisia, bromelain, papain, turfgrass, caprolactone bromide, carbocysteine, eprinodone, mesna, ambroxol, sobrerol, domiodol, letosteine, stepronin, tiopronin, gelsolin, nucleosin β4, dornase alfa, neltenexin, and erdosteine), and DNase (e.g., rhDNase). The surface modifier can be provided within the nanoparticle and / or on the surface of the nanoparticle composition (e.g., by coating, adsorption, covalent bonding, or other methods).

[0242] The nanoparticle composition may further comprise one or more functionalized lipids. For example, the lipid may be functionalized with an alkynyl group that can undergo cycloaddition reaction when exposed to azide under appropriate reaction conditions. In particular, lipid bilayers may be functionalized with one or more groups in this manner to facilitate membrane permeation, cell recognition, or imaging (imaging). The surface of the nanoparticle composition may also be conjugated with one or more useful antibodies. Functional groups and conjugates useful for targeted cell delivery, imaging, and membrane permeation are well known in the art.

[0243] In addition to the above components, the nanoparticle composition may contain any substance useful in pharmaceutical compositions. For example, the nanoparticle composition may contain one or more pharmaceutically acceptable excipients or auxiliary ingredients, such as, but not limited to, one or more solvents, dispersion media, diluents, dispersion aids, suspending agents, granulation aids, disintegrants, fillers, glidants, liquid carriers, binders, surfactants, isotonicity agents, thickeners or emulsifiers, buffers, lubricants (lubricants), oils, preservatives, and other materials. Excipients such as waxes, butters, colorants, coating agents, flavors, and flavoring agents may also be included. Pharmaceutically acceptable excipients are well known in the art.

[0244] Examples of diluents may include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and / or combinations thereof. Granulating and dispersing agents may be selected from potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate, sodium lauryl sulfate, quaternary ammonium compounds and / or combinations thereof.

[0245] Surfactants and / or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, chondroitin, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and VEEGUM® [magnesium aluminum silicate]), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol), triacetin monostearate, ethylene glycol distearate, glycerol monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene ethers), sorbitan esters (e.g., PEG-100 ester ... Polyoxyethylene sorbitan monolaurate [TWEEN® 20], polyoxyethylene sorbitan [TWEEN® 60], polyoxyethylene sorbitan monooleate [TWEEN® 80], sorbitan monopalmitate [SPAN® 40], sorbitan monostearate [SPAN® 60], sorbitan tristearate [SPAN® 65], glycerol monooleate, sorbitan monooleate [SPAN® 80], polyoxyethylene esters (e.g., polyoxyethylene ethylene monostearate [MYRJ® 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate and SOLUTOL®), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., CREMOPHOR®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether [BRIJ® 30]), poly(vinylpyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate,These include, but are not limited to, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium dodecyl sulfate, PLURONIC® F68, POLOXAMER® 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and / or combinations thereof.

[0246] Binders may be starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol), natural and synthetic gums (e.g., acacia, sodium alginate, Irish moss extract, panwar gum, ghatti gum, isabel shell mucilage, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate (VEEGUM®), and larch arabinogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts; silicic acid, polymethacrylates, waxes, water, alcohol, combinations thereof, or any other suitable binder.

[0247] Examples of preservatives may include, but are not limited to, antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcoholic preservatives, acidic preservatives, and / or other preservatives. Examples of antioxidants include, but are not limited to, α-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and / or sodium sulfite. Examples of chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and / or trisodium edetate. Examples of antibacterial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bromophenol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerol, hexyloxybutanol, imidurea, phenethyl alcohol, phenylmercuric nitrate, propylene glycol, and / or thimerosal. Examples of antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and / or sorbic acid. Examples of alcoholic preservatives include, but are not limited to, ethanol, polyethylene glycol, benzyl alcohol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and / or phenylethyl alcohol. Examples of acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroascorbic acid, ascorbic acid, sorbic acid, and / or phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopheryl acetate, deoxyoxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, GLYDANTPLUS®, PHENONIP®, methyl p-hydroxybenzoate, GERMALL® 115, GERMABEN® II, NEOLONE™, KATHON™, and / or EUXYL®.

[0248] Examples of buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, d-gluconic acid, calcium glycerophosphate, calcium lactate, calcium lactobionate, propionic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydrogen phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate mixtures, tromethamine, sulfamate buffer, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and / or combinations thereof. Lubricants are selected from magnesium stearate, calcium stearate, stearic acid, silicon dioxide, talc, malt, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, and magnesium laurate, sulfates, sodium lauryl sulfate, and combinations thereof.

[0249] Examples of oils include almond, apricot kernel, avocado, babassu, bergamot, blackcurrant seed, borage, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba oil, lavender, lavandin, lemon, macadamia nut, mallow, mango seed, mink skin, nutmeg, olive, orange, palm, palm kernel, and peach floss. Ingredients include, but are not limited to, corn, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, essence, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, elderberry, vetiver, walnut, and wheat germ oils, as well as butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and / or combinations thereof.

[0250] The nanoparticle compositions described herein can include a lipid component and one or more other components, such as a therapeutic agent and / or a prophylactic agent. The nanoparticle compositions can be designed for one or more specific uses or targets. The components of the nanoparticle composition can be selected based on the specific use or target and / or based on the efficacy, toxicity, cost, ease of use, availability, or other characteristics of one or more components. Similarly, a particular formulation of the nanoparticle composition can be selected for a particular use or target based, for example, on the efficacy and toxicity of a particular combination of components.

[0251] The lipid component of the nanoparticle composition may be, for example, any of the following: (I), (IA), (IA-1), (IA-2), (IA-3), (IA-4), (IA-5), (IA-6), (IIA), (IIA-A), (IIA-B), (IIA-C), (IIA-D), (IIA-E), (IIA-F), (IIA-G), (IIA-H), (IB), (IB-1), (IB-2), (IB-3), (IB-4), (IB-5), (IB-6), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IB-5), (IB-6), (IC), (IC-1), (IC-2), (IC-3), (IC-4), (IC-5), (IC-6), (IC ... 2), (IC-3), (IC-4), (IC-5), (IC-6), (IC-7), (IC-8), (IC-9), (IC-10), (IC-11), (ID), (ID-1), (ID-2), (ID-3), (ID-4), (ID-5), (ID-6), (ID-7), (ID-8), (ID-9), (ID-10), (ID-11), phospholipids (e.g., unsaturated lipids such as DOPE or DSPC), PEG lipids, and structured lipids.

[0252] Nanoparticle compositions can be designed for one or more specific applications or targets. For example, nanoparticle compositions can be designed to deliver therapeutic and / or prophylactic agents, such as RNA, to specific cells, tissues, organs, or systems, or groups thereof, within a mammalian body. The physicochemical properties of the nanoparticle composition can be modified to enhance selectivity for specific bodily targets. For example, particle size can be adjusted based on the window sizes of different organs. The therapeutic and / or prophylactic agents included in the nanoparticle composition can also be selected based on one or more desired delivery targets. For example, therapeutic and / or prophylactic agents can be selected for a specific indication, condition, disease, or disorder, and / or for delivery (e.g., localized or specific delivery) to specific cells, tissues, organs, or systems, or groups thereof. In certain embodiments, nanoparticle compositions can include mRNA encoding a polypeptide of interest, which can be translated intracellularly to produce the polypeptide of interest. Such compositions can be designed for specific delivery to a specific organ. In some embodiments, the composition can be designed for specific delivery to the mammalian liver.

[0253] The amount of therapeutic and / or prophylactic agent in a nanoparticle composition can vary depending on the size, composition, desired target and / or use, or other characteristics of the nanoparticle composition, and the nature of the therapeutic and / or prophylactic agent. For example, the amount of RNA that can be used in a nanoparticle composition can vary depending on the size, sequence, and other characteristics of the RNA. The relative amounts of therapeutic and / or prophylactic agent and other components (e.g., lipids) in the nanoparticle composition can also vary.

[0254] The properties of nanoparticle compositions can vary depending on their components. For example, a nanoparticle composition containing cholesterol as a structural lipid can have different properties from a nanoparticle composition containing a different structural lipid. Similarly, the properties of nanoparticle compositions can vary depending on the absolute or relative amounts of their components. For example, a nanoparticle composition containing a relatively high molar fraction of phospholipids can have different properties from a nanoparticle composition containing a relatively low molar fraction of phospholipids. The properties can also vary depending on the preparation method and conditions of the nanoparticle composition.

[0255] Nanoparticle compositions 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 nanoparticle compositions. Dynamic light scattering or potentiometry (e.g., potentiometric titration) can be used to measure the zeta potential. Dynamic light scattering can also be used to determine particle size.

[0256] The nanoparticle composition has an average particle size of 50 nm to 110 nm.

[0257] The nanoparticle composition can be relatively homogeneous. The polydispersity index can be used to indicate the homogeneity of the nanoparticle composition, e.g., the particle size distribution of the nanoparticle composition. A small polydispersity index (e.g., less than 0.3) generally indicates a narrow particle size distribution. The nanoparticle composition has a polydispersity index of 0.04 to 0.20. For example, the polydispersity index can be 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, or 0.19.

[0258] The encapsulation efficiency of a therapeutic and / or prophylactic agent represents the amount of therapeutic and / or prophylactic agent encapsulated in or otherwise associated with a nanoparticle composition after preparation, relative to the amount initially provided. It is expected that the encapsulation efficiency will be as high as possible (e.g., approaching 100%). For example, the encapsulation efficiency can be measured by comparing the amount of therapeutic and / or prophylactic agent in a solution containing the nanoparticle composition before and after dissolving the nanoparticle composition with one or more organic solvents or detergents. Fluorescence can be used to measure the amount of free therapeutic and / or prophylactic agent (e.g., RNA) in solution. For the nanoparticle compositions described herein, the encapsulation efficiency of a therapeutic and / or prophylactic agent can be at least 50%, e.g., 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the encapsulation efficiency may be at least 80%. In certain embodiments, the encapsulation efficiency may be at least 90%.

[0259] The nanoparticle composition can optionally include one or more coatings. For example, the nanoparticle composition can be formulated into a coated capsule, film, or tablet. The capsule, film, or tablet containing the composition described herein can be of any useful size, tensile strength, hardness, or density.

[0260] The nanoparticle compositions of the present application can be formulated, in whole or in part, as pharmaceutical compositions. Pharmaceutical compositions can include one or more nanoparticle components. For example, a pharmaceutical composition can include one or more nanoparticle compositions containing one or more different therapeutic and / or prophylactic agents. Pharmaceutical compositions can further include one or more pharmaceutically acceptable excipients or auxiliary ingredients, such as those described herein, so long as any conventional excipient or auxiliary ingredient is not incompatible with one or more of the components of the nanoparticle composition. If an excipient or auxiliary ingredient is incompatible with a component of the nanoparticle composition, its combination with that component may result in undesirable biological or other adverse effects.

[0261] In some embodiments, one or more excipients or auxiliary ingredients may comprise more than 50% of the total mass or volume of a pharmaceutical composition containing a nanoparticle composition. For example, one or more excipients or auxiliary ingredients may comprise 50%, 60%, 70%, 80%, 90%, or more of a pharmaceutically customary amount. In some embodiments, a pharmaceutically acceptable excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipient is approved for human and veterinary use.

[0262] The relative amounts of one or more nanoparticle compositions, one or more pharmaceutically acceptable excipients, and / or any other ingredients in a pharmaceutical composition according to the present application will vary depending on the identity, size, and / or disease state of the subject being treated, as well as the route of administration of the composition. By way of example, a pharmaceutical composition may contain one or more nanoparticle compositions in the range of 0.1% to 100% (w / w).

[0263] Nanoparticle compositions and / or pharmaceutical compositions comprising one or more of such nanoparticle compositions can be administered to any patient or subject, including those who can benefit from a therapeutic effect provided by delivery of a therapeutic and / or prophylactic agent to one or more specific cells, tissues, organs, or systems or groups thereof (e.g., the renal system). While the description of nanoparticle compositions and pharmaceutical compositions comprising nanoparticle compositions provided herein primarily relates to compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to other mammals. Modifications to compositions suitable for administration to humans are well known for making compositions suitable for administration to a variety of animals, and such modifications can often be designed and / or performed by skilled veterinary pharmacologists through no more than routine (if any) experimentation, including but not limited to humans, other primates, and other artificially created animals, including commercially relevant mammals such as cows, pigs, horses, sheep, cats, dogs, rats, and / or mice.

[0264] Pharmaceutical compositions containing one or more nanoparticle compositions can be prepared by any method known or hereafter developed in the art of pharmacology. Generally, such preparative methods involve combining an active ingredient with an excipient and / or one or more other accessory ingredients, and then dividing, shaping, and / or packaging the product into desired single- or multi-dose units, as desired or necessary.

[0265] Pharmaceutical compositions according to the present application can be manufactured, packaged, and / or sold in bulk, in single unit doses, and / or in multiple single unit doses. As used herein, a "unit dose" is a precise amount of pharmaceutical composition containing a predetermined amount of active ingredient (e.g., nanoparticle composition). The amount of active ingredient is approximately equal to the dose of the active ingredient that would be administered to a subject and / or a convenient fraction of that dose, e.g., one-half or one-third of that dose.

[0266] Pharmaceutical compositions can be prepared in various forms suitable for various routes and methods of administration. For example, pharmaceutical compositions can be prepared in liquid dosage forms (e.g., emulsions, microemulsions, nanoemulsions, solutions, suspensions, syrups, and pharmaceutical preparations), injectable dosage forms, solid dosage forms (e.g., capsules, tablets, lozenges, etc.), liquids, powders, and granules), preparations for topical and / or transdermal administration (e.g., ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and patches), suspensions, powders, and other forms. Liquid dosage forms for oral and parenteral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, nanoemulsions, solutions, suspensions, syrups, and / or drugs.

[0267] In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol, and mixtures thereof. In addition to inert diluents, oral compositions may also contain other therapeutic and / or prophylactic agents, other medicinal agents, such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents, and / or fragrances. In certain embodiments for parenteral administration, the composition is mixed with a solubilizing agent (e.g., alcohol, oil, modified oil, glycol, polysorbate, cyclodextrin, polymer, and / or combination thereof).

[0268] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersants, wetting agents, and / or suspending agents. Sterile injectable preparations can be sterile injectable solutions, suspensions, and / or emulsions in non-toxic parenterally acceptable diluents and / or solvents, for example, solutions in 1,3-butanediol. Acceptable vehicles and solvents, including water, can be used. Sterile, fixed oils are conventionally used as solvents or suspending media. For this purpose, any mild, fixed oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids, such as oleic acid, can be used in the preparation of injectables.

[0269] Injectable preparations can be sterilized, for example, by filtration through a bacteria-retaining filter, and / or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0270] In order to prolong the effect of an active ingredient, it is often desirable to slow the absorption of the active ingredient from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. In this case, the rate of absorption of the drug depends upon its rate of dissolution, which, in turn, depends upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form can be accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers (such as polylactide-polyethylene glycol). The rate of drug release can be controlled depending on the drug-to-polymer ratio and the properties of the particular polymer used. Examples of other biodegradable polymers include poly(ortho-esters) and poly(anhydrides). Stock injectable formulations are prepared by incorporating the drug in liposomes or microemulsions that are compatible with body tissues.

[0271] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the composition with a suitable non-irritating excipient such as cocoa butter, polyethylene glycol or a suppository wax which is solid at ordinary temperatures but liquid at body temperature and which melts in the rectum or vaginal cavity and releases the active ingredient.

[0272] Solid dosage forms for oral administration include capsules, tablets, pills, films, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient, such as sodium citrate or dicalcium phosphate and / or fillers or extenders (e.g., starches, lactose, sucrose, glucose, mannitol, and silicic acid), binders (e.g., carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia), humectants (e.g., glycerol), disintegrating agents (e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate), delaying agents (e.g., paraffin), absorption accelerators (e.g., quaternary ammonium compounds), wetting agents (e.g., cetyl alcohol and glyceryl monostearate), absorbents (e.g., kaolin and bentonite, silicates), and lubricants (e.g., talc), calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0273] Similar types of solid compositions can be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation field.These may optionally contain opacifying agents and may have a composition that releases the active ingredient only, or preferably in a certain part of the intestinal tract, optionally delayed release.Examples of embedding compositions that can be used include polymeric substances and waxes.Similar types of solid compositions can be used as fillers in soft and hard-filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycol.

[0274] Dosage forms for topical and / or transdermal administration of the composition may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and / or patches. Typically, the active ingredient is mixed under sterile conditions with a pharmaceutically acceptable excipient and / or any preservatives and / or buffers that may be required. Furthermore, the present application contemplates the use of transdermal patches, which generally offer the added advantage of controlled delivery of the compound into the body. Such dosage forms can be prepared, for example, by dissolving and / or dispersing the compound in a suitable medium. Alternatively or additionally, the rate can be controlled by providing a rate-controlling membrane and / or by dispersing the compound in a polymer matrix and / or gel.

[0275] Formulations suitable for topical administration include, but are not limited to, liquid and / or semi-liquid formulations, such as liniments, lotions, oil-in-water and / or water-in-oil emulsions, such as creams, ointments and / or pastes, and / or solutions and / or suspensions. However, the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further include one or more of the other ingredients described herein.

[0276] Pharmaceutical compositions can be prepared, packaged, and / or sold as formulations suitable for pulmonary administration via the buccal cavity. Such formulations can contain dry particles of the active ingredient. Such compositions are conveniently in dry powder form for administration using a device containing a dry powder reservoir into which a stream of propellant can be directed to disperse the powder, and / or using a self-propelling solvent / powder dispensing container (such as a device containing the active ingredient dissolved and / or suspended in a low-boiling propellant in a sealed container). Powder compositions may contain a solid fine powder diluent, such as sugar, and are conveniently provided in a unit dose form.

[0277] Low-boiling propellants typically include liquid propellants having a boiling point below 65°F at atmospheric pressure. Typically, the propellant may comprise 50% to 99.9% (w / w) of the composition, and the active ingredient may comprise 0.1% to 20% (w / w) of the composition. The propellant may further comprise other ingredients, such as a liquid nonionic and / or solid anionic surfactant and / or a solid diluent (the particle size of which may be on the same order as the particles containing the active ingredient).

[0278] Pharmaceutical compositions formulated for pulmonary delivery can provide the active ingredient in the form of droplets of a solution and / or suspension. Such formulations can be prepared, packaged, and / or sold as aqueous and / or dilute alcoholic solutions and / or suspensions (optionally sterile) containing the active ingredient, and can be conveniently administered using any spray and / or nebulizing device. Such formulations may further contain one or more other ingredients, including, but not limited to, flavorings such as sodium saccharin, volatile oils, buffers, surfactants, and / or preservatives such as methyl hydroxybenzoate. The average diameter of the droplets provided by this route of administration can be in the range of 1 nm to 200 nm.

[0279] The formulations described herein useful for pulmonary delivery can also be used for intranasal delivery of pharmaceutical compositions. Another formulation suitable for intranasal administration is a coarse powder containing the active ingredient and having an average particle size of 0.2 to 500 μm. Such a formulation is administered in snuff form by rapid inhalation through the nasal passages from a powder container held close to the nose.

[0280] Formulations suitable for nasal administration may contain, for example, 0.1% (w / w) to 100% (w / w) of the active ingredient, and may also contain one or more other active ingredients described herein. Pharmaceutical compositions may also be prepared, packaged, and / or sold in a formulation suitable for buccal administration. Such formulations may be, for example, in the form of tablets and / or lozenges prepared using conventional methods and may contain, for example, 0.1% to 20% (w / w) of the active ingredient, with the remainder comprising an orally dissolvable and / or degradable composition, and optionally one or more other ingredients described herein. Alternatively, formulations suitable for oral administration may comprise a powder and / or aerosolized and / or atomized solution and / or suspension containing the active ingredient. Such powdered, aerosolized, and / or aerosolized formulations, when dispersed, have an average particle size and / or droplet size in the range of 0.1 nm to 200 nm and may further comprise one or more other ingredients described herein.

[0281] Pharmaceutical compositions can be prepared, packaged, and / or sold in a formulation suitable for ocular administration. Such formulations can be, for example, in the form of drops, comprising, for example, a 0.1 / 1.0% (w / w) solution and / or suspension of the active ingredient in an aqueous or oily liquid vehicle. Such drops can further comprise one or more buffering agents, salts, and / or other additive ingredients described herein. Other formulations useful for ocular administration include formulations comprising the active ingredient in microcrystalline and / or liposomal form. Ear drops and / or eye drops are contemplated as being within the scope of the present disclosure.

[0282] Method for producing polypeptides in cells The present application further provides a method for producing a polypeptide of interest in mammalian cells. The method for producing a polypeptide includes contacting the cells with a nanoparticle composition containing mRNA encoding the polypeptide of interest. After contacting the cells with the nanoparticle composition, the mRNA can be taken up into the cells and translated within the cells to produce the polypeptide of interest.

[0283] Generally, the step of contacting mammalian cells with a nanoparticle composition containing mRNA encoding a polypeptide of interest can be performed in vivo, ex vivo, in culture, or in vitro. The amount of nanoparticle composition contacted with cells and / or the amount of mRNA therein can vary depending on the type of cell or tissue contacted, the mode of administration, the physicochemical properties (e.g., size, charge, and chemical composition) of the nanoparticle composition and mRNA, and other factors. Generally, an effective amount of the nanoparticle composition can efficiently produce the polypeptide in the cells. Measures of efficiency can include polypeptide translation (indicated by polypeptide expression), mRNA degradation levels, and immune response indicators.

[0284] The step of contacting a cell with a nanoparticle composition containing mRNA may involve or cause transfection. Phospholipids contained in the lipid component of the nanoparticle composition can facilitate and / or increase the efficiency of transfection, for example, by interacting with and / or fusing with cellular or intracellular membranes. Transfection allows for translation of the mRNA within the cell.

[0285] In some embodiments, the nanoparticle compositions described herein can be used in therapy. For example, the mRNA contained in the nanoparticle composition can encode a therapeutic polypeptide (e.g., in a translatable region) and produce the therapeutic polypeptide upon contact with and / or entry into a cell (e.g., transfection). In other embodiments, the mRNA contained in the nanoparticle composition can encode a polypeptide that can improve or increase immunity in a subject. For example, the mRNA can encode granulocyte colony-stimulating factor or trastuzumab.

[0286] In certain embodiments, the mRNA contained in the nanoparticle composition may encode a recombinant polypeptide that can replace one or more polypeptides that may be substantially absent in cells contacted with the nanoparticle composition. The one or more substantially absent polypeptides may be absent due to genetic mutations in the encoding gene or its regulatory pathway. Alternatively, the recombinant polypeptide produced by translation of the mRNA may antagonize the activity of an endogenous protein present within the cell, present on the cell surface, or secreted from the cell. Antagonistic recombinant polypeptides may be desirable to counteract adverse effects caused by the activity of endogenous proteins, such as altered activity or localization caused by mutations. Alternatively, the recombinant polypeptide produced by translation of the mRNA may indirectly or directly antagonize the activity of a biological moiety present within the cell, present on the cell surface, or secreted from the cell. Antagonistic biological moieties include, but are not limited to, lipids (e.g., cholesterol), lipoproteins (e.g., low-density lipoprotein), nucleic acids, carbohydrates, and small molecule toxins. The recombinant polypeptide produced by translation of the mRNA can be engineered to be localized intracellularly (e.g., in a specific compartment such as the nucleus), or can be engineered to be secreted from the cell, or can be engineered to be transported to the plasma membrane of the cell.

[0287] In some embodiments, contacting cells with a nanoparticle composition containing mRNA can reduce the cellular innate immune response to an exogenous nucleic acid. Cells can be contacted with a first nanoparticle composition containing a first amount of a first exogenous mRNA containing a translatable region, and the level of the cellular innate immune response to the first exogenous mRNA can be determined. Subsequently, the cells can be contacted with a second composition containing a second amount of the first exogenous mRNA, the second amount of the first exogenous mRNA being less than the first amount. Alternatively, the second composition can contain a first amount of a second exogenous mRNA that is different from the first exogenous mRNA. The step of contacting cells with the first and second compositions can be repeated one or more times. Furthermore, the efficiency of polypeptide production (e.g., translation) in the cells can optionally be determined, and the cells can be repeatedly re-contacted with the first and / or second compositions until a target protein production efficiency is achieved.

[0288] The present application further provides methods for treating a disease or disorder in a mammal, comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition described in any one of the preceding paragraphs, wherein the nanoparticle composition is particularly useful for treating diseases, disorders, or conditions characterized by lost or abnormal protein or polypeptide activity. For example, a nanoparticle composition containing mRNA encoding a missing or abnormal polypeptide can be administered or delivered to a cell. Subsequent translation of the mRNA can produce the polypeptide, thereby reducing or eliminating the problem caused by the lack or abnormality of the polypeptide's activity. Because translation can occur rapidly, these methods and compositions can be useful for treating acute diseases, disorders, or conditions (e.g., sepsis, stroke, and myocardial infarction). Therapeutic and / or prophylactic agents contained in the nanoparticle composition can also affect gene expression by altering the transcription rate of a given species.

[0289] Diseases, disorders and / or conditions characterized by dysfunctional or aberrant protein or polypeptide activity include, but are not limited to, rare diseases, infectious diseases (e.g., vaccines and therapeutics), cancer and proliferative diseases, genetic diseases (e.g., cystic fibrosis), autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renovascular diseases, and metabolic diseases.

[0290] The present application provides methods involving administering nanoparticle compositions comprising one or more therapeutic and / or prophylactic agents, as well as pharmaceutical compositions comprising nanoparticle compositions. The terms therapeutic and prophylactic are used interchangeably herein with respect to features and embodiments of the present application. Therapeutic compositions or imaging, diagnostic, or prophylactic compositions thereof can be administered to a subject using any reasonable amount and any route of administration effective for preventing, treating, diagnosing, or imaging a disease, disorder, and / or condition, and / or for any other purpose. The specific amount administered to a particular subject may vary depending on the subject's species, age, and general condition; the purpose of administration; the specific ingredients; the mode of administration, etc. The compositions according to the present application can be formulated in unit dosage form for ease of administration and uniformity of dosage. However, it will be understood that the total daily amount of the compositions of the present application will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective, prophylactically effective, or other appropriate dosage level (e.g., for imaging) for any particular patient will vary depending on a variety of factors, including the severity and identification (if any) of the disease being treated; the one or more therapeutic and / or prophylactic agents used; the particular components used; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and excretion rate of the particular pharmaceutical component used; the duration of treatment; drugs used in combination with or concurrently with the particular pharmaceutical component; and factors well known in the medical arts.

[0291] The present application further provides a method for specifically delivering a therapeutic and / or prophylactic agent to a mammalian organ, comprising administering to the mammal a nanoparticle composition described in any one of the preceding paragraphs, wherein the administration comprises contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the organ. The therapeutic and / or prophylactic agent, such as a protein, a cytotoxic agent, a radioactive ion, a chemotherapeutic agent, or a nucleic acid (e.g., RNA, e.g., mRNA), can be delivered to a cell or organ. If the therapeutic and / or prophylactic agent is mRNA, upon contact of the cell with the nanoparticle composition, translatable mRNA can be translated within the cell to produce a polypeptide of interest. However, substantially non-translatable mRNA can also be delivered to a cell. Substantially non-translatable mRNA can be used as a vaccine and / or can sequester cellular translation components to reduce the expression of other species within the cell.

[0292] In some embodiments, the nanoparticle composition can be targeted to a particular type or class of cells (e.g., cells of a particular organ or system). For example, a nanoparticle composition containing a therapeutic and / or prophylactic agent of interest can be specifically delivered to the liver, kidney, spleen, femur, gastrointestinal tract, or lung of a mammal. Specific delivery to a particular class of cells, organ, or system, or group thereof, means, for example, that when the nanoparticle composition is administered to a mammal, a higher proportion of the nanoparticle composition containing the therapeutic and / or prophylactic agent is delivered to the targeted destination (e.g., tissue) compared to other destinations. In some embodiments, the target tissue is selected from the liver, kidney, lung, spleen, femur, eye, gastrointestinal tissue (e.g., via intraocular, subretinal, or intravitreal injection), blood vessels (e.g., intracoronary or intrafemoral) or vascular endothelium of the kidney, and tumor tissue (e.g., via intratumoral injection).

[0293] As another example of targeted or specific delivery, mRNA encoding a cell surface protein-binding partner (e.g., an antibody or functional fragment thereof, a scaffold protein, or a peptide) or receptor can be included in the nanoparticle composition. The mRNA can alternatively be increased to direct the synthesis and extracellular localization of lipids, carbohydrates, or other biological moieties. Alternatively, other therapeutic and / or prophylactic agents or components of the nanoparticle composition (e.g., lipids or ligands) can be selected based on their affinity for a particular receptor (e.g., low-density lipoprotein receptor) so that the nanoparticle composition can more readily interact with a target cell population containing the receptor.

[0294] In some embodiments, the ligand may be a surface-bound antibody, which may allow for modulation of cell target specificity. This is particularly useful because it allows for the raising of highly specific antibodies at desired target sites against epitopes of interest. In one embodiment, multiple antibodies are expressed on the cell surface, and each antibody may have a different specificity for the desired target. Such an approach may improve the affinity and specificity of targeted interactions.

[0295] Target cells may include, but are not limited to, hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, lung cells, bone cells, stem cells, mesenchymal cells, neural cells, cardiac cells, adipocytes, vascular smooth muscle cells, cardiac muscle cells, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, and tumor cells.

[0296] In some embodiments, the nanoparticle composition may target liver cells.

[0297] In some embodiments, the nanoparticle composition can target extrahepatic cells.

[0298] In some preferred embodiments, the nanoparticle compositions may be targeted to the spleen, stomach, or intestinal tract.

[0299] The nanoparticle compositions of the present application can be administered by any route. In some embodiments, compositions, including prophylactic, diagnostic, or imaging compositions comprising one or more nanoparticle compositions of the present application, are administered by one or more of a variety of routes, including orally, intravenously, intramuscularly, intraarterially, intramedullary, intrathecally, subcutaneously, intracerebroventricularly, transdermally or intradermally, rectally, intravaginally, intraperitoneally, intraocularly, subretinally, intravitreally, topically (e.g., via powders, ointments, creams, gels, lotions, and / or drops), mucosally, nasally, bucally, enterally, vitreally, intratumorally, sublingually, intranasally; by intratracheal instillation, bronchial instillation, and / or inhalation; as an oral spray and / or powder, a nasal spray and / or aerosol, and / or via a portal vein catheter. In some embodiments, the compositions can be administered intravenously, intramuscularly, intradermally, intraarterially, intratumorally, subcutaneously, intraocularly, subretinally, intravitreally, or by inhalation. However, in view of possible advances in the science of drug delivery, the present application contemplates delivery or administration of the compositions described herein by any suitable route. Generally, the most appropriate route of administration will depend on various factors, including the properties of the nanoparticle composition comprising one or more therapeutic and / or prophylactic agents (e.g., stability in various body environments (e.g., blood and gastrointestinal tract)), the condition of the patient (e.g., whether the patient can tolerate a particular route), etc.

[0300] In certain embodiments, the therapeutic and / or prophylactic agent is administered to a mammal at a dose of 0.0001 mg / kg to 10 mg / kg, e.g., 0.0001 mg / kg to 10 mg / kg, 0.001 mg / kg to 10 mg / kg, 0.005 mg / kg to 10 mg / kg, 0.01 mg / kg to 10 mg / kg, 0.05 mg / kg to 10 mg / kg, 0.1 mg / kg to 10 mg / kg, 1 mg / kg to 10 mg / kg, 2 mg / kg to 10 mg / kg, 5 mg / kg to 10 mg / kg, 0.0001 mg / kg to 5 mg / kg, 0.001 mg / kg~5mg / kg, 0.005mg / kg~5mg / kg, 0.01mg / kg~5mg / kg, 0.05mg / kg~5mg / kg, 0.1mg / kg~5mg / kg, 1mg / kg~5mg / kg , 2mg / kg~5mg / kg, 0.0001mg / kg~2.5mg / kg, 0.001mg / kg~2.5mg / kg, 0.005mg / kg~2.5mg / kg, 0.01mg / kg~2.5mg / kg 0.05mg / kg~2.5mg / kg, 0.1mg / kg~2.5mg / kg, 1mg / kg~2.5mg / kg, 2mg / kg~2.5mg / kg, 0.0001 mg / kg~1mg / kg, 0.001mg / kg~1mg / kg, 0.005mg / kg~1mg / kg, 0.01mg / kg~1mg / kg, 0.05mg / kg~ 1mg / kg, 0.1mg / kg~1mg / kg, 0.0001mg / kg~0.25mg / kg, 0.001mg / kg~0.25mg / kg, 0.005mg / kg It can be ~0.25mg / kg, 0.01mg / kg~0.25mg / kg, 0.05mg / kg~0.25mg / kg or 0.1mg / kg~0.25mg / kg. In some embodiments, a therapeutic and / or prophylactic dose of a nanoparticle composition (e.g., mRNA) can be administered in the range of about 0.001 mg / kg to about 10 mg / kg. In other embodiments, a therapeutic and / or prophylactic dose of about 0.005 mg / kg to about 2.5 mg / kg can be administered. In certain embodiments, a dose of about 0.1 mg / kg to about 1 mg / kg can be administered. In other embodiments, a dose of about 0.05 mg / kg to about 0.25 mg / kg can be administered.To achieve a desired level of mRNA expression and / or therapeutic, diagnostic, preventive, or imaging effect, doses can be administered one or more times daily in the same or different amounts. The desired dose can be administered, for example, three times a day, twice a day, once a day, once every other day, once every three days, once a week, once every two weeks, once every three weeks, or once every four weeks. In certain embodiments, multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations) can be used to deliver the desired dose. In some embodiments, a single dose can be administered, for example, before or after surgery or in the case of an acute disease, disorder, or condition.

[0301] Nanoparticle compositions containing one or more therapeutic and / or prophylactic agents can be used in combination with one or more other therapeutic, prophylactic, diagnostic, or imaging agents. "In combination with" does not imply that the agents must be administered simultaneously and / or formulated for delivery together, although such delivery methods are within the scope of this application. For example, one or more nanoparticle compositions containing one or more different therapeutic and / or prophylactic agents can be administered in combination. The compositions can be administered simultaneously with, prior to, or after one or more other desired therapeutic or medical treatments. Typically, each agent is administered at a dose and / or schedule determined for that agent. In some embodiments, this application encompasses delivering the composition or its imaging, diagnostic, or prophylactic composition in combination with an agent that enhances its bioavailability, reduces and / or alters its metabolism, inhibits its excretion, and / or modulates contraction in vivo.

[0302] It is understood that therapeutic, prophylactic, diagnostic, or imaging active agents used in combination can be administered together in a single composition or can be administered separately in different compositions. In general, it is desirable to use agents used in combination at levels that do not exceed the levels at which they are used individually. In some embodiments, the levels used in combination may be lower than the levels at which they are used individually.

[0303] The particular combination of therapies (treatments or procedures) to use in a combination regimen will take into account the compatibility of the desired therapies and / or procedures, as well as the desired therapeutic effect to be achieved. It will also be understood that the therapies employed may achieve a desired effect for the same condition (e.g., a composition used to treat cancer may be administered simultaneously with a chemotherapeutic agent) or may achieve a different effect (e.g., control of any side effects, such as infusion-related reactions).

[0304] In some embodiments, methods of treating a subject in need of treatment or methods of delivering a therapeutic and / or prophylactic drug to a subject (e.g., a mammal) can include pretreating the subject with one or more agents prior to administering the nanoparticle composition. For example, the subject can be pretreated with an effective amount (e.g., 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, or any other effective amount) of dexamethasone, methotrexate, acetaminophen, an H1 receptor blocker, or an H2 receptor blocker. Pretreatment can occur within 24 hours (e.g., 24 hours, 20 hours, 16 hours, 12 hours, 8 hours, 4 hours, 2 hours, 1 hour, 50 minutes, 40 minutes, 30 minutes, 20 minutes, or 10 minutes) of administration of the nanoparticle composition, and can occur, for example, once, twice, or more times with increasing doses.

[0305] It is to be understood that the various embodiments described herein (including the examples below) and features in the various embodiments can be combined with one another in any combination, and the various embodiments resulting from these combinations are included within the scope of this application in the same way as if the embodiments resulting from these combinations were specifically and individually set forth herein, unless the context clearly dictates otherwise. [Brief explanation of the drawings]

[0306] [Figure 1] 1 shows the results of luciferase expression of LNPs prepared based on compounds of the present invention 24 hours after intramuscular injection. [Figure 2A] 1 shows the tissue and organ distribution of luciferase in LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Figure 2B] 1 shows the tissue and organ distribution of luciferase in LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Figure 2C] 1 shows the tissue and organ distribution of luciferase in LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Figure 2D] 1 shows the tissue and organ distribution of luciferase in LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Figure 3A] 1 shows the liver ALT / AST levels of LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Figure 3B] 1 shows the liver ALT / AST levels of LNPs prepared based on compounds of the present invention 24 hours after intravenous injection. [Example]

[0307] The following examples are illustrative of the present application and are not intended to limit the present application in any way. Unless otherwise indicated, all temperatures are in degrees Celsius and pressures are at or near atmospheric pressure. All reagents and starting materials used in this application are commercially available except as synthetic intermediates or where otherwise indicated.

[0308] Example 1 Synthesis of Compound 1

[0309] [ka]

[0310] The synthetic route is shown below:

[0311] [ka]

[0312] Specifically, compound B1 (1.9 g, 7.47 mmol), DCC (1.8 g, 8.96 mmol), and DMAP (437 mg, 3.58 mmol) were added to a solution of compound A1 (2.0 g, 8.96 mmol) in dichloromethane (30 mL). The reaction mixture was stirred at room temperature for 7 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to give compound C1 (2.9 g, 85.00%).

[0313] To a solution of compound E1 (5.0 g, 25.63 mmol) in dichloromethane (60 mL), compound D1 (3.7 g, 21.36 mmol), DCC (5.29 g, 25.63 mmol), and DMAP (1.2 g, 10.25 mmol) were added. The reaction mixture was stirred at room temperature for 7 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to give compound F1 (6.5 g, 88.00%).

[0314] To a solution of compound F1 (500 mg, 1.44 mmol) in DMF (15 mL), compound G (562 mg, 2.88 mmol), NBu4I (795 mg, 2.15 mmol), and K2CO3 (597 mg, 4.32 mmol) were added. The reaction mixture was stirred at room temperature for 12 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (10 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to give compound H1 (580 mg, 87% yield).

[0315] To a solution of compound H1 (500 mg, 1.08 mmol) in DMF (15 mL), compound C1 (745 mg, 1.62 mmol), NBu4I (598 mg, 1.62 mmol), and Cs2CO3 (1.4 g, 4.32 mmol) were added. The reaction mixture was stirred at room temperature for 15 h and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (10 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to give compound I1 (811 mg, 89% yield).

[0316] To a solution of compound I1 (10 g, 11.85 mmol) in dichloromethane (100 mL), concentrated hydrochloric acid (60 mL) was added, and the reaction mixture was stirred at room temperature for 3 hours and monitored by TLC. After completion of the reaction, the layers were separated. The organic layer was washed with saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate=30:1) to give compound J1 (6.5 g, 81.00%).

[0317] To a solution of compound J1 (1 g, 1.48 mmol) in toluene (25 mL), compound K1 (470 mg, 4.44 mmol) and pyridinium 4-methylbenzenesulfonate (744 mg, 2.96 mmol) were added and heated to reflux for 20 h using a Dean-Stark apparatus. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, extracted with ethyl acetate (10 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give compound L1 (648 mg, 57% yield).

[0318] To a solution of compound L1 (750 mg, 0.98 mmol) in ultra-dry dichloromethane (10 mL), methanesulfonic anhydride (340 mg, 1.96 mmol) and anhydrous triethylamine (0.45 mL) were added at 0°C and stirred for 12 hours. The reaction was monitored by TLC. After completion of the reaction, the reaction solution was cooled to room temperature, diluted with an equal amount of water, and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give compound M1 (800 mg, yield 97%).

[0319] Compound M1 (750 mg, 0.89 mmol) was added to a solution of dimethylamine (2 M in THF) (30 mL), sealed, and stirred at room temperature for 6 days. The reaction was monitored by TLC. After completion of the reaction, the mixture was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to give compound 1 (212 mg, 30% yield) as an oil. 1H NMR(600MHz,CDCl3)δ 4.87-4.82(m,1H),4.05-4.02(m,2H),3.90-3.85(m,2H),3.59-3.56(m,2H),2.31-2.23(m,4H),2.21-2.19( m,8H),1.94-1.87(brs,1H),1.68-1.56(m,10H),1.52-1.42(m,4H),1.36-1.21(m,52H),0.87-0.84(m,9H); 13 C NMR(151MHz,CDCl3)δ 173.98,173.77,100.76,74.22,64.55,62.87,62.81,59.34,59.31,45.97,3 5.37,35.28,34.86,34.49,34.46,34.28,32.47,32.39,32.03,31.99,29.99 29.72,29.65,29.46,29.36,28.80,26.06,25.44,25.29,25.16,23.49,23.26,23.16,22.92,22.79,14.22;MS-ESI(m / z):794.6(M+H) + .

[0320] Example 2 Synthesis of Compound 2

[0321] [ka]

[0322] The preparation method was the same as that of Compound 1, except that 1,2,4-butanetriol was used as raw material instead of Compound K1 to produce Compound 2 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.82-4.78(m,1H),4.04-3.96(m,4H),3.42-3.39(m,1H),2.39-2.34(brs,1H),2.29-2.25(brs,1H),2.23-2.18 (m,10H),1.77-1.71(brs,1H),1.66-1.65(brs,1H),1.58-1.39(m,13H),1.33-1.16(m,53H),0.82-0.80(m,9H); 13C NMR(151MHz,CDCl3)δ 173.62,173.41,111.82,74.63,74.61,73.88,69.89,64.25,56.18,45.28,37.73,37.4 8,37.40,37.20,34.59,34.19,34.18,34.09,31.83,31.78,31.60,29.70,29.68,29.52, 29.50,29.45,29.34,29.32,29.25,29.20,29.15,29.09,29.07,28.59,25.86,25.23,25 .07,24.89,23.88,23.58,23.56,23.24,22.60,22.58,14.01;MS-ESI(m / z):794.6(M+H) + .

[0323] Example 2A Synthesis of Compound 2A

[0324] [ka]

[0325] The preparation method was the same as that of Compound 1, except that S-1,2,4-butanetriol was used as raw material instead of Compound K1 to produce Compound 2A as an oil. 1 H NMR(400MHz,CDCl3)δ 4.97-4.83(m,1H),4.17-4.05(m,4H),3.52(t,J=8.0Hz,1H),2.58-2.37(m,2H),2.35-2.23 (m,10H),1.91-1.70(m,2H),1.69-1.50(m,14H),1.48-1.24(m,52H),0.92(t,J=8.0Hz,9H); 13C NMR(100MHz,CDCl3)δ 174.04,173.69,112.10,74.59,74.12,69.94,64.34,56.22,45.21,37.79,37. 70,37.44,37.38,34.74,34.42,34.16,31.91,31.88,31.43,31.41,29.80,29. 57,29.55,29.52,29.49,29.32,29.28,29.25,29.19,28.63,28.61,25.95,25. 33,25.17,25.00,23.89,23.71,23.59,22.68,14.12;MS-ESI(m / z):795.0(M+H) + .

[0326] Example 3 Synthesis of Compound 3

[0327] [ka]

[0328] The preparation method was the same as that of Compound 1, except that 3-dimethylamino-1,2-propanediol and p-toluenesulfonic acid monohydrate were used as raw materials instead of Compound K1 and pyridinium 4-methylbenzenesulfonate to produce Compound 3 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.85-4.81(m,1H),4.20-4.16(m,1H),4.06-4.01(m,3H),3.49-3.46(m,1H),2.48-2.44(m,1H),2.35-2.32 (m,1H),2.27-2.22(m,10H),1.60-1.54(m,8H),1.48-1.47(m,4H),1.31-1.20(m,54H),0.86-0.83(m,9H); 13C NMR(151MHz,CDCl3)δ 173.90,173.69,112.58,74.40,74.36,74.13,69.06,64.48,62.62,62.59,46.22,37.80, 37.55,37.51,37.30,34.79,34.40,34.38,34.24,31.99,31.94,29.88,29.83,29.68,29. 66,29.61,29.58,29.52,29.47,29.41,29.38,29.33,29.31,29.27,29.25,28.74,26.01, 25.39,25.24,25.06,25.06,24.03,23.70,23.35,22.74,14.17;MS-ESI(m / z):780.6(M+H) + .

[0329] Example 4 Synthesis of Compound 4

[0330] [ka]

[0331] The preparation method was the same as that of Compound 1, except that undecanoic acid and 6-bromo-1-hexanol were used as raw materials instead of Compound D1 and Compound E1 to produce Compound 4 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.86-4.81(m,1H),4.04-4.01(m,2H),3.90-3.84(m,2H),3.59-3.55(m,2H),2.27-2.22(m,4H),2.20-2.17( m,8H),1.94-1.85(brs,1H),1.67-1.55(m,10H),1.48-1.45(m,4H),1.36-1.22(m,52H),0.86-0.83(m,9H); 13C NMR(151MHz,CDCl3)δ 174.01,173.70,100.73,74.14,64.40,64.36,62.83,62.75,59.31,59.28,45.98,35. 50,35.25,34.79,34.47,34.24,32.45,32.43,32.22,31.98,31.95,29.94,29.70,29. 64,29.61,29.59,29.55,29.42,29.39,29.35,29.32,29.29,29.25,28.73,26.05,25. 40,25.24,25.10,23.45,23.32,23.21,23.08,22.75,14.18;MS-ESI(m / z):794.6(M+H) + .

[0332] Example 5 Synthesis of Compound 5

[0333] [ka]

[0334] The preparation method was the same as that of Compound 1, except that undecanoic acid, 6-bromo-1-hexanol, and 1,2,4-butanetriol were used as raw materials instead of Compound D1, Compound E1, and Compound K1 to produce Compound 5 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.81(p,J=6.2Hz,1H),4.03(t,J=6.6Hz,1H),3.99(t,J=6.6Hz,3H),3.42(t,J=7.6Hz,1H),2.41-2.36(brs,1H),2.31-2.26(brs,1 H),2.23-2.20(m,10H),1.78-1.73(brs,1H),1.66-1.60(brs,1H),1.58-1.43(m,12H),1.31-1.20(m,54H),0.82(t,J=7.0Hz,9H); 13C NMR(151MHz,CDCl3)δ 173.92,173.60,112.00,74.75,74.73,74.05,70.00,64.31,56.29,45.41,37.83, 37.73,37.50,37.42,34.73,34.40,34.20,31.93,31.90,31.73,31.71,29.82,29. 80,29.58,29.56,29.53,29.50,29.33,29.30,29.26,29.20,28.67,28.65,25.96, 25.35,25.19,25.05,23.90,23.71,23.59,22.69,14.13;MS-ESI(m / z):794.6(M+H) + .

[0335] Example 5A Synthesis of Compound 5A

[0336] [ka]

[0337] The preparation method was the same as that of Compound 1, except that undecanoic acid, 6-bromo-1-hexanol, and S-1,2,4-butanetriol were used as raw materials instead of Compound D1, Compound E1, and Compound K1 to produce Compound 5A as an oil. 1 H NMR(400MHz,CDCl3)δ 4.97-4.83(m,1H),4.17-4.06(m,4H),3.51(t,J=4.0Hz,1H),2.51-2.45(m,1H),2.44-2.32(m,1H),2.31-2.23 (m,10H),1.87-1.80(m,1H),1.74-1.67(m,1H),1.66-1.50(m,14H),1.48-1.24(m,52H),0.92(t,J=8.0Hz,9H); 13C NMR(100Hz,CDCl3)δ 173.96,173.62,111.98,74.71,74.08,69.96,64.30,56.24,45.36,37.60,3 7.53,37.50,37.43,34.71,34.39,34.13,31.87,31.84,31.67,31.61,29.78, 29.70,29.51,29.48,29.30,29.28,29.24,29.18,28.64,28.61,25.94,25.32 ,25.17,25.03,23.89,23.70,23.59,22.67,14.11;MS-ESI(m / z):795.0(M+H) + .

[0338] Example 5B Synthesis of Compound 5B

[0339] [ka]

[0340] The preparation method was the same as that of Compound 1, except that undecanoic acid, 6-bromo-1-hexanol, and R-1,2,4-butanetriol were used as raw materials instead of Compound D1, Compound E1, and Compound K1 to produce Compound 5B as an oil. 1 H NMR(400MHz,CDCl3)δ 4.97-4.83(m,1H),4.17-4.05(m,4H),3.51(t,J=4.0Hz,1H),2.51-2.45(m,1H),2.44-2.32(m,1H),2.31-2.23 (m,10H),1.86-1.80(m,1H),1.74-1.67(m,1H),1.66-1.51(m,14H),1.48-1.24(m,52H),0.92(t,J=8.0Hz,9H); 13C NMR(100MHz,CDCl3)δ 173.97,173.62,111.98,74.71,74.08,69.96,64.30,56.24,45.36,37.60,3 7.43,37.50,37.44,34.71,34.39,34.13,31.87,31.84,31.67,31.61,29.79, 29.70,29.51,29.48,29.30,29.28,29.24,29.18,28.64,28.61,25.94,25.32 ,25.18,25.03,23.89,23.70,23.59,22.67,14.11;MS-ESI(m / z):795.0(M+H) + .

[0341] Example 6 Synthesis of Compound 6

[0342] [ka]

[0343] The preparation method was the same as that of Compound 1, except that undecanoic acid, 6-bromo-1-hexanol, and 3-dimethylamino-1,2-propanediol were used as raw materials instead of Compound D1, Compound E1, and Compound K1 to produce Compound 6 as an oil. 1 H NMR(500MHz,CDCl3)δ 4.84(p,J=6.2Hz,1H),4.19(p,J=6.5Hz,1H),4.06-4.01(m,3H),3.49(t,J=7.8Hz,1H),2.48(dd,J=12.6,6.8Hz,1H),2.35(d d,J=12.6,5.0Hz,1H),2.27-2.23(m,10H),1.62-1.53(m,10H),1.49-1.46(m,4H),1.33-1.24(m,52H),0.85(t,J=6.8Hz,9H); 13C NMR (126 MHz, CDCl3) δ 174.05,173.72,112.66,74.44,74.18,69.08,64.40,62.65,46.25,37.8 2,37.71,37.49,37.44,34.83,34.50,34.26,32.00,31.97,29.91,29.86, 29.63,29.61,29.40,29.37,29.33,29.27,28.73,26.04,25.42,25.26,25 .12,24.06,23.95,23.73,23.61,22.77,14.19;MS-ESI(m / z):780.6(M+H) + .

[0344] Example 7 Synthesis of Compound 7

[0345] [ka]

[0346] The preparation method was the same as that of Compound 1, except that 8-bromo-1-octanol, 2-octyl-decanoic acid, and 3-dimethylamino-1,2-propanediol were used as raw materials instead of Compound A1, Compound B1, and Compound K1 to produce Compound 7 as an oil. 1 H NMR(500MHz,CDCl3)δ 4.22-4.17(m,1H),4.06-4.01(m,5H),3.48(t,J=7.8Hz,1H),2.50-2.46(m,1H),2.39-2.36(m,1H),2 .28-2.24(m,9H),1.60-1.53(m,13H),1.42-1.37(m,2H),1.30-1.22(m,53H),0.85(t,J=6.8Hz,9H); 13C NMR(126MHz,CDCl3)δ 176.64,173.92,112.62,74.26,74.21,68.96,64.29,64.10,62.48,46.07, 45.85,37.75,37.60,37.42,37.32,34.40,32.56,31.90,31.87,29.91,29. 86,29.57,29.46,29.31,29.25,29.18,28.75,28.64,27.47,25.99,25.94, 25.03,23.98,23.85,23.64,23.51,22.67,14.10;MS-ESI(m / z):794.6(M+H) + .

[0347] Example 8 Preparation of Compound 8

[0348] [ka]

[0349] The preparation method was the same as that of Compound 1, except that 8-bromo-1-octanol, 2-octyl-decanoic acid and 1,2,4-butanetriol were used as raw materials instead of Compound A1, Compound B1 and Compound K1 to produce Compound 8 as an oil. 1 H NMR(500MHz,CDCl3)δ 4.07-4.00(m,6H),3.44(t,J=7.5Hz,1H),2.42-2.36(brs 1H),2.32-2.21(m,10H),1.81-1.74(brs,1H),1.68-1.51(m,14H),1.42-1.34(brs,2H),1.26-1.22(m,53H),0.84(t,J=6.9Hz,9H); 13C NMR (126 MHz, CDCl3) δ 176.69,173.97,112.05,74.81,70.06,64.36,64.16,56.36,45.92,45. 50,37.93,37.78,37.59,37.48,34.46,32.63,31.97,31.94,31.82,29. 63,29.53,29.37,29.32,29.24,28.81,28.71,27.54,26.06,26.02,25. 09,24.07,23.94,23.77,23.64,22.73,14.16;MS-ESI(m / z):808.6(M+H) + .

[0350] Example 9 Preparation of Compound 9

[0351] [ka]

[0352] The preparation method was the same as that of Compound 1, except that 8-bromo-1-octanol, 2-octyl-decanoic acid, undecanoic acid, and 6-bromo-1-hexanol were used as raw materials instead of Compounds A1, B1, D1, and E1 to produce Compound 9 as an oil. 1 H NMR(500MHz,CDCl3)δ 4.04-4.01(m,4H),3.89-3.85(m,2H),3.59-3.56(m,2H),2.31-2.18(m,11H),1.9 3-1.86(brs,1H),1.67-1.52(m,12H),1.43-1.23(m,56H),0.85(t,J=6.9Hz,9H); 13C NMR(126MHz,CDCl3)δ 176.71,174.00,100.75,64.40,64.36,64.18,62.83,62.76,59.34,59.31F,4 5.98,45.94,35.46,35.25,34.48,32.64,32.54,32.46,32.34,31.98,31.96, 30.06,29.71,29.65,29.55,29.39,29.34,29.26,28.83,28.74,27.56,26.08 ,25.11,23.46,23.32,23.23,23.09,22.75,14.18;MS-ESI(m / z):808.5(M+H) + .

[0353] Example 10 Preparation of Compound 10

[0354] [ka]

[0355] The preparation method was the same as that of Compound 1, except that 1,2,4-butanetriol and N,N-ethylmethylamine were used as raw materials instead of Compound K1 and dimethylamine to produce Compound 10 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.83-4.75(m,1H),4.05-3.93(m,4H),3.42-3.35(m,1H),2.46-2.30(m,4H),2.23-2.15(m,7H),1.79-1.70(m,1H),1.67- 1.59(m,1H),1.56-1.51(m,6H),1.43(d,J=7.2Hz,4H),1.29-1.14(m,56H),0.99(t,J=7.1Hz,3H),0.80(t,J=7.0Hz,9H); 13C NMR(151MHz,CDCl3)δ 173.72,173.51,111.83,74.85,74.81,73.96,69.99,64.33,53.70,51.43,41.49,37.80, 37.56,37.48,37.28,34.66,34.26,34.25,34.15,31.90,31.85,31.30,29.77,29.59,29. 57,29.51,29.49,29.41,29.38,29.32,29.29,29.24,29.22,29.16,29.13,28.65,25.92, 25.30,25.13,24.97,23.96,23.66,23.31,22.65,14.08,12.09;MS-ESI(m / z):808.7(M+H) + .

[0356] Example 11 Preparation of Compound 11

[0357] [ka]

[0358] The preparation method was the same as that of Compound 1, except that 1,2,4-butanetriol and diethylamine were used as raw materials instead of Compound K1 and dimethylamine to produce Compound 11 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.86-4.80(m,1H),4.08-4.00(m,4H),3.49-3.43(m,1H),2.77-2.62(m,7H),2.25(dt,J=10.5,7.5Hz,4H),1.85-1.75 (m,2H),1.63-1.56(m,6H),1.50-1.44(m,4H),1.37-1.19(m,56H),1.12(t,J=7.2Hz,6H),0.85(td,J=7.1,2.4Hz,9H); 13C NMR(151MHz,CDCl3)δ 173.95,173.73,112.22,74.59,74.55,74.17,69.94,64.52,49.36,46.95,37.84,37. 59,37.42,37.21,34.80,34.39,34.24,31.99,31.95,30.30,29.87,29.69,29.67,29.6 2,29.59,29.50,29.47,29.42,29.35,29.32,29.29,29.26,28.74,26.02,25.40,25.24 ,25.07,24.09,23.82,23.76,23.47,22.75,14.20,10.74;MS-ESI(m / z):822.9(M+H)+.

[0359] Example 12 Preparation of Compound 12

[0360] [ka]

[0361] For the preparation of compound 12, refer to the preparation of compound 2.

[0362] Example 13 Preparation of Compound 13

[0363] [ka]

[0364] The preparation method was the same as that of Compound 1, except that 1,2,5-pentanetriol was used as raw material instead of Compound K1 to produce Compound 13 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.81-4.74(m,1H),4.00-3.92(m,4H),3.38-3.33(m,1H),2.25-2.17(m,6H),2.15(s,6H) ,1.59-1.46(m,15H),1.46-1.37(m,5H),1.29-1.12(m,50H),0.79(td,J=7.1,2.3Hz,9H); 13C NMR(151MHz,CDCl3)δ 173.69,173.47,111.86,76.08,76.05,73.92,69.93,69.91,F64.29,59.48,53.38,45.24,37.8 0,37.56,37.47,37.25,34.64,34.24,34.22,34.13,31.88,31.83,31.20,29.76,29.57,29.55, 29.49,29.47,29.39,29.37,29.30,29.27,29.22,29.20,29.14,29.11,28.63,25.90,25.28,25 .12,24.95,23.95,23.89,23.64,23.62,23.29,22.64,22.62,14.05;MS-ESI(m / z):808.8(M+H) + .

[0365] Example 14 Preparation of Compound 14

[0366] [ka]

[0367] The preparation method was the same as that of Compound 1, except that 1,2,6-hexanetriol was used as raw material instead of Compound K1 to produce Compound 14 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.91-4.79(m,1H),4.02(m,4H),3.45-3.34(m,1H),2.33(t,J=6.5Hz,2H),2.27 (m,10H),1.60(m,9H),1.53-1.41(m,8H),1.25(m,55H),0.87(t,J=7.0Hz,9H); 13C NMR (151 MHz, chloroform-d) δ 111.89, 76.18, 74.09, 70.03, 64.42, 59.48, 45.20, 37.94-37.16 (m), 34.73, 34.33, 34.15, 33.31, 31.88, 29.81, 29.59, 29.51, 29.44, 29.32, 29.24, 28.66, 27.37, 25.92, 25.31, 25.16, 24.99, 23.68, 22.66, 14.09; MS-ESI (m / z): 822.9 (M+H) + .

[0368] Example 15 Preparation of Compound 15

[0369] [ka]

[0370] The preparation method was the same as that of Compound 1, except that 1,2,4-butanetriol and N-methyl-n-propylamine were used as raw materials instead of Compound K1 and dimethylamine to produce Compound 15 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.84-4.78(m,1H),4.07-3.97(m,4H),3.42(t,J=7.7Hz,1H),2.53-2.39(m,2H),2.33(t,J=7.6Hz,2H),2.26-2.20(m,7H),1.8 1-1.74(m,1H),1.74-1.66(m,1H),1.61-1.54(m,6H),1.51-1.41(m,5H),1.35-1.14(m,55H),0.84(dt,J=15.0,7.3Hz,12H).; 13C NMR(151MHz,CDCl3)δ 173.86,173.64,111.97,74.82,74.78,74.09,70.02,64.45,59.66,54.18,41.99,37.87,3 7.62,37.52,37.32,34.76,34.35,34.22,31.97,31.92,31.06,29.85,29.83,29.64,29.58, 29.56,29.49,29.46,29.39,29.31,29.29,29.25,29.22,28.72,25.99,25.37,25.21,25.04 ,24.05,23.75,23.72,23.41,22.73,22.71,20.09,14.15,11.84;MS-ESI(m / z):822.8(M+H) + .

[0371] Example 16 Preparation of Compound 16

[0372] [ka]

[0373] The preparation method was the same as that of Compound 1, except that 1,2,4-butanetriol and N,N-ethylisopropylamine were used as raw materials instead of Compound K1 and dimethylamine to produce Compound 16 as an oil. 1 H NMR(600MHz,CDCl3)δ 4.84(p,J=6.3Hz,1H),4.11-4.01(m,4H),3.47(t,J=7.6Hz,1H),3.34-2.50(m ,5H),2.29-2.22(m,4H),1.64-1.44(m,14H),1.40-1.03(m,63H),0.86(t,9H); 13C NMR(151MHz,CDCl3)δ 173.95,173.73,112.18,74.58,74.19,69.97,64.54,46.37,44.49,37.86,37.61,37.4 4,37.23,34.82,34.41,34.25,32.01,31.97,29.90,29.87,29.80,29.70,29.68,29.63, 29.60,29.50,29.43,29.36,29.33,29.28,28.76,26.03,25.42,25.25,25.10,25.08,24 .12,23.85,23.79,23.50,22.78,22.76,18.33,17.19,14.20;MS-ESI(m / z):836.8(M+H) +

[0374] Example 17 Preparation of Compound 17

[0375] [ka]

[0376] Specifically, compound B2 (5.0 g, 19.57 mmol), EDCI (4.9 g, 25.44 mmol), and DMAP (0.96, 7.83 mmol) were added to a solution of compound A1 (5.24 g, 23.49 mmol) in dichloromethane (30 mL). The reaction mixture was stirred at room temperature for 7 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to give compound C2 (7.19 g, 80.00%).

[0377] To a solution of compound D2 (7.0 g, 40.8 mmol) in dichloromethane (60 mL), compound E1 (9.6 g, 49.0 mmol), EDCI (10.2 g, 53.1 mmol), and DMAP (2.0 g, 16.4 mmol) were added. The reaction mixture was stirred at room temperature for 7 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to give compound F2 (12.20 g, 86.15%).

[0378] To a solution of compound F2 (12.2 g, 35.14 mmol) in DMF (50 mL), compound G1 (13.72 g, 70.28 mmol), NBu4I (13.0 g, 35.14 mmol), and K2CO3 (14.57 g, 105.43 mmol) were added. The reaction mixture was stirred at room temperature for 12 h and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (50 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give compound H2 (9.9 g, 61.11% yield).

[0379] To a solution of compound H2 (9.9 g, 21.42 mmol) in DMF (100 mL), compound C2 (11.19 g, 33.12 mmol), NBu4I (8.0 g, 21.24 mmol), and Cs2CO3 (14 g, 42.84 mmol) were added. The reaction mixture was stirred at room temperature for 15 h and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (100 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to give compound I2 (12 g, 66.67% yield).

[0380] To a solution of compound I2 (12 g, 14.26 mmol) in dichloromethane (100 ml), concentrated hydrochloric acid (60 ml) was added, and the reaction mixture was stirred at room temperature for 3 hours and monitored by TLC. After the reaction was completed, the layers were separated. The organic layer was washed with saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1) to give compound G2 (7.72 g, 80.00%).

[0381] To a solution of compound G2 (3 g, 4.44 mmol) in toluene (25 mL), compound K2 (1.42 g, 13.32 mmol) and pyridinium 4-methylbenzenesulfonate (1.34 g, 6.65 mmol) were added. The reaction mixture was heated to reflux for 20 h using a Dean-Stark apparatus and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, extracted with ethyl acetate (50 mL x 3), concentrated, and purified by silica gel column chromatography (dichloromethane:methanol = 40:1) to give compound L2 (1.3 g, 43.33% yield).

[0382] To a solution of compound L2 (1.3 g, 1.7 mmol) in ultra-dry dichloromethane (10 mL), methanesulfonic anhydride (0.59 g, 3.4 mmol) and anhydrous triethylamine (0.7 mL) were added at 0 °C. The reaction mixture was stirred for 12 hours and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal amount of water, and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 40:1) to give compound M2 (1.32 g, 92.30% yield).

[0383] Compound M2 (1.32 g, 1.57 mmol) was added to a solution of 30 mL of dimethylamine (2 M in THF), and the reaction mixture was sealed and stirred at room temperature for 6 days. The reaction mixture was monitored by TLC. After completion of the reaction, the mixture was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 30:1 + aqueous ammonia) to give compound 17 (300 mg, 24% yield) as an oil.1 H NMR(500MHz,クロロホルム-d)δ 5.84-5.69(m,1H),5.34(t,J=9.1Hz,1H),4.05-3.96(m,2H),3.88-3.79(m,1H ),3.41(t,J=7.4Hz,1H),3.16(q,J=6.7Hz,2H),2.44-2.33(m,1H),2.29-2.22( m,1H),2.19(d,J=3.3Hz,6H),2.09(t,J=7.6Hz,4H),1.79-1.69(m,1H),1.66-1 .47(m,8H),1.45-1.37(m,3H),1.30-1.17(m,56H),0.82(t,J=6.9,2.4Hz,9H); 13 C NMR(126MHz,CDCl3)δ 172.97,172.62,111.98,111.94,74.72,74.68,69.97,56.31,49.09,45.48,45.4 5,39.53,37.71,37.49,37.38,37.25,37.11,36.74,36.72,35.33,31.93,31.90, 31.75,29.73,29.63,29.59,29.48,29.36,29.30,29.24,26.99,25.99,25.94,25 .80,25.76,23.92,23.77,23.60,23.40,22.69,14.14;MS-ESI(m / z):792.9(M+H)+

[0384] Example 18 Compound 18

[0385]

change

[0386] The synthetic path is as follows:

[0387]

change

[0388] Specifically, compound B1 (10 g, 39.02 mmol), EDCI (14.96 g, 78.04 mmol), and DMAP (1.90 g, 15.61 mmol) were added to a solution of compound A1 (10.40 g, 46.82 mmol) in dichloromethane (100 mL). The reaction mixture was stirred at room temperature for 7 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to obtain compound C1 (15 g, 83.52%).

[0389] To a solution of compound C1 (15 g, 32.60 mmol) in DMF (100 mL), compound G1 (2.12 g, 10.86 mmol), NBu4I (4 g, 10.86 mmol), and Cs2CO3 (10.62 g, 32.6 mmol) were added. The reaction mixture was stirred at room temperature for 12 h and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (100 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 40:1) to give compound I3 (7.95 g, 76.60% yield).

[0390] To a solution of compound I3 (7.95 g, 8.31 mmol) in dichloromethane (100 mL), concentrated hydrochloric acid (60 mL) was added, and the reaction mixture was stirred at room temperature for 3 hours and monitored by TLC. After the reaction was complete, the layers were separated. The organic layer was washed with saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate=50:1) to give compound J3 (5.89 g, 89.65%).

[0391] To a solution of compound J3 (5.89 g, 7.45 mmol) in toluene (25 mL), compound K2 (2.4 g, 22.35 mmol) and pyridinium 4-methylbenzenesulfonate (2.95 g, 11.73 mmol) were added. The reaction mixture was heated to reflux for 20 h using a Dean-Stark apparatus and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, extracted with ethyl acetate (50 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give compound L3 (2.94 g, 44.95% yield).

[0392] To a solution of compound L3 (2.94 g, 3.34 mmol) in ultra-dry dichloromethane (10 mL), methanesulfonic anhydride (1.2 g, 6.68 mmol) and anhydrous triethylamine (0.8 mL) were added at 0 °C. The reaction mixture was stirred for 12 hours and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, and separated. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give compound M3 (3.1 g, 97% yield).

[0393] Compound M3 (3.1 g, 3.24 mmol) was added to a solution of 30 mL of dimethylamine (2 M in THF), and the reaction mixture was sealed and stirred at room temperature for 6 days. The reaction mixture was monitored by TLC. After completion of the reaction, the mixture was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 40:1) to give compound 18 (439 mg, 14.98% yield) as an oil. 1H NMR(500MHz,CDCl3)δ 4.87-4.79(m,2H),4.08-3.98(m,2H),3.44(t,J=7.4Hz,1H),2.46-2.37(m,1H),2.36-2.27(m,1H),2.22(t,J=6.7Hz,10H), 1.83-1.74(m,1H),1.72-1.62(m,1H),1.58-1.51(m,3H),1.46(d,J=6.1Hz,8H),1.36-1.14(m,69H),0.84(t,J=6.9Hz,12H); 13C NMR(126MHz,CDCl3)δ 173.69,112.11,74.73,74.10,70.02,56.33,45.42,37.85,37.53,34.78,34.23,31.94,31.70,29.87 ,29.60,29.31,29.27,29.24,25.39,25.23,24.05,23.75,22.74,14.18;MS-ESI(m / z):906.9(M+H)+.

[0394] Example 19 Compound 19

[0395] [ka]

[0396] The preparation method was the same as that of Compound 18, except that 6-bromohexanoic acid and undecanol were used as raw materials instead of Compound A1 and Compound B1 to produce Compound 19 as an oil. 1 H NMR(500MHz,CDCl3)δ 4.08-3.98(m,6H),3.44(t,J=7.4Hz,1H),2.46-2.38(m,1H),2.37-2.29(m,1H),2.29-2.20(m,10H), 1.83-1.74(m,1H),1.72-1.65(m,1H),1.61-1.52(m,11H),1.29-1.22(m,41H),0.85(t,J=6.9Hz,6H); 13 C NMR(126MHz,CDCl3)δ 173.94,111.93,74.78,70.07,64.50,56.33,45.44,37.67,37.39,34.39,31.99,31.67,29.68,29.6 1,29.50,29.42,29.34,28.74,26.01,25.06,23.75,23.42,22.77,14.21;MS-ESI(m / z):628.6(M+H)+

[0397] Example 20A Compound 20A

[0398] [ka]

[0399] The synthetic route is shown below:

[0400] [ka]

[0401] Specifically, compound B4 (0.25 g, 1.25 mmol), DCC (0.28 g, 1.38 mmol), and DMAP (0.01 g, 0.06 mmol) were added to a solution of compound A1 (0.28 g, 1.25 mmol) in dichloromethane (30 mL). The reaction mixture was stirred at room temperature for 2 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of saturated sodium bicarbonate solution and separated. The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to obtain compound C4 (0.50 g, 88.90%).

[0402] To a solution of compound C4 (0.50 g, 1.23 mmol) in DMF (5 mL) was added compound G1 (0.12 g, 0.62 mmol), NBuI (0.05 g, 0.12 mmol), and CsCO (1.21 g, 3.71 mmol). The reaction mixture was stirred at room temperature for 12 hours and monitored by TLC. After completion of the reaction, the reaction mixture was diluted with an equal volume of water, extracted with ethyl acetate (10 mL × 3), and concentrated to give crude compound I4, which was used directly in the next loading step.

[0403] To a solution of compound I4 (crude, 1.23 mmol) in dichloromethane (10 mL), concentrated hydrochloric acid (5 mL) was added, and the reaction mixture was stirred at room temperature for 3 hours and monitored by TLC. After completion of the reaction, the layers were separated. The organic layer was washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:1) to give compound J4 (0.27 g, 32.27% yield over two steps).

[0404] To a solution of compound J4 (0.27 g, 0.40 mmol) in toluene (25 mL), compound K2 (0.17 g, 1.59 mmol) and pyridinium 4-methylbenzenesulfonate (0.10 g, 0.02 mmol) were added. The reaction mixture was heated to reflux for 20 h using a Dean-Stark apparatus and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, extracted with ethyl acetate (10 mL x 3), concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give compound L4 (0.28 g, 91.79% yield).

[0405] To a solution of compound L4 (0.28 g, 0.36 mmol) in dichloromethane (10 mL), methanesulfonic anhydride (0.10 g, 0.55 mmol) and anhydrous triethylamine (0.20 mL) were added, and the reaction was stirred at room temperature for 2 hours and monitored by TLC. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with an equal volume of water, and separated. The organic layer was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated to give crude compound M4, which was used directly in the next loading step.

[0406] Compound M4 (crude, 0.36 mmol) was added to a solution of 20 mL of dimethylamine (2 M in THF) under nitrogen protection, and the reaction mixture was stirred at room temperature for 3 days and monitored by TLC. After completion of the reaction, the mixture was concentrated and purified by silica gel column chromatography (dichloromethane:methanol = 30:1) to give compound 20A as an oil (0.21 g, 66.50% yield in two steps). 1H NMR(400MHz,CDCl3)δ 4.20-4.09(m,6H),3.55(t,J=8.0Hz,1H),2.87-2.69(m,2H),2.55(s,6H),2.32(t,J=8.0Hz ,4H),1.94-1.87(m,2H),1.68-1.56(m,12H),1.36-1.21(m,50H),0.92(t,J=8.0Hz,9H);13C NMR(400MHz,CDCl3)δ 173.96,112.52,73.98,69.66,62.94,55.97,44.30,37.66,37.17,34.63,34.42,33.57,32.49,32.23,29.73 29.71,29.27,29.14,26.21,24.98,23.97,23.71,22.66,14.09;MS-ESI(m / z):795.0(M+H)+.

[0407] Example 21 Compound 21

[0408] [ka]

[0409] The preparation method was the same as that of Compound 18, except that 6-bromohexanol and 2-hexyldecanoic acid were used as raw materials instead of Compound A1 and Compound B1 to produce Compound 21 as an oil. 1H NMR(600MHz,chloroform-d)δ 4.11-4.02(m,6H),3.47(t,J=7.6Hz,1H),2.47-2.41(brs,1H),2.37-2.33(brs,1H),2.32-2.24(m,8H),1.84-1.78(brs,1H),1. 72-1.65(brs,1H),1.63-1.54(m,12H),1.45-1.40(m,4H),1.37-1.31(m,10H),1.30-1.21(m,42H),0.87(td,J=7.0,1.3Hz,12H); 13 C NMR (600 MHz, chloroform-d) δ 176.70, 111.95, 74.72, 69.98, 64.08, 56.24, 45.84, 45.34, 37.75, 37.43, 32.52, 31.85, 31.69, 31.62, 29.56, 29.44, 29.24, 29.22, 28.69, 27.46, 27.41, 25.99, 23.90, 23.60, 22.66, 22.58, 14.07; MS-ESI (m / z): 823.0 (M+H)+.

[0410] Example of test Experimental Example 1: Lipid Nanoparticle (LNP) Encapsulation The mRNA stock solution was dispersed in 20 mM acetic acid solution (pH 5.0) to a final concentration of 200 μg / mL (aqueous phase). A mixture of compound, cholesterol, DSPC, and DMG-PEG2000 (50:38.5:10:1.5 molar ratios shown in Table 1) was dissolved in absolute ethanol to form a lipid mixture (oil phase). The mRNA was mixed with the lipid mixture by T-mixing, and the flow rates of the aqueous and oil phases (3:1) were controlled to a total flow rate of 12–50 mL / min to obtain LNP-encapsulated mRNA. The encapsulated LNP was diluted with buffer (pH 5.0–5.5) and then concentrated by ultrafiltration while exchanging the diluent. Finally, the mRNA LNP was concentrated to 200–600 μg / mL, and the pH of the LNP was adjusted to approximately 7–8. Finally, the total and free mRNA content in the LNPs was detected using the Quant-iT™ RiboGreen™ RNA Assay Kit (Invitogen, Thermo Fisher Scientific, Cat. No. R11490) and 10% Triton X-100 solution or 10% OTG solution as an emulsion breaker, and the encapsulation efficiency of the LNPs was calculated. The final LNP product was diluted with diluent, and then 1 mL was added to a particle size cell and loaded onto a Malvern ZetaSizer instrument to detect the particle size of the LNPs. The results are shown in Table 1.

[0411] [Table 2]

[0412] Experimental Example 2: Toxicity Assay Nanoparticle samples containing different lipids were intravenously administered at a dose of 3 mg / kg to rats (SD rats, Zhejiang Vital River Laboratory Animal Technology Co., Ltd., 3 males and 3 females per group, body weight: females 220-300 g, males 260-340 g). 24 hours after administration, the expression of various clinical markers was evaluated and compared with that induced by an SM102 LNP formulation (i.e., an LNP formulation made from a known SM102 lipid at the same ratio, prepared according to the method in Example 1) and phosphate-buffered saline (PBS).

[0413] [Table 3]

[0414] The results shown in Table 2 show that the values ​​of aspartate aminotransferase and alanine aminotransferase in the group of the compound of the present invention are obviously reduced compared to the group of SM102, which indicates that the toxicity of the compound of the present invention is further reduced, suggesting that the compound of the present invention has higher biological safety.

[0415] Experimental Example 3: Delivery and expression effects of the lipid nanoparticle composition of the present invention Five BALB / c mice (Zhejiang Vital River Laboratory Animal Technology Co., Ltd., female, 6-8 weeks old, 18-20 g) were assigned to each group, and their tails were numbered 1# to 5# with a black marker pen. Five mice in each group were sequentially intramuscularly injected with 5 μg of lipid nanoparticle composition. The lipid nanoparticle composition of the present invention was prepared according to Test Example 1 and encapsulated with luciferase mRNA. An MC3 LNP group was set up in parallel as a reference (i.e., an LNP formulation made from the known MC3 lipid obtained by preparing according to the method and ratios in Test Example 1).

[0416] Fluorescence assays were performed on mice from each group 24 hours after intramuscular administration of the sample (fluorescein sodium salt YEASEN, LOT: D5330150, IP, 150 mg / kg, administered 10 minutes prior to administration to each group). After anesthetization, the mice were placed in the IVIS® Lumina LT Series III small animal bio-optical imaging system, in the order of tail marks 1# to 5#. Whole-body fluorescence was detected to evaluate the delivery and expression of the formulated luciferase mRNA.

[0417] The LNP delivery and expression effects of the compounds of the present invention are shown in Figure 1, relative to the expression of MC3. The data demonstrate that the compounds of the present invention have excellent delivery and expression effects. The compounds of the present invention achieved delivery and expression effects equivalent to or even superior to those of MC3. In particular, the delivery and expression effects of Compound 2, Compound 2A, Compound 5, Compound 5A, Compound 5B, Compound 8, Compound 13, Compound 14, and Compound 20A were 3 to 4 times greater than those of MC3. The results for Compounds 2, 2A, Compound 5, Compound 5A, and Compound 5B clearly demonstrate that differences in composition do not affect the delivery activity of the pH-responsive lipids of the present invention.

[0418] Experimental Example 4: Assay of extrahepatic delivery effect of lipid compounds of the present invention Each group consisted of five BALB / c mice (Zhejiang Vital River Laboratory Animal Technology Co., Ltd., female, 6-8 weeks old, 18-20 g) whose tails were numbered 1# to 5# with a black marker pen. Five mice in each group were intravenously injected with 20 μg of lipid nanoparticle composition. The lipid nanoparticle composition of the present invention was prepared according to Test Example 1, and luciferase mRNA was encapsulated within it. A phosphate-buffered saline (PBS) group was run in parallel as a blank control, and the MC3 group (prepared according to the method and ratios in Test Example 1) was run as a reference.

[0419] Fluorescence assays were performed on mice from each group 24 hours after intravenous administration of the sample (fluorescein sodium salt YEASEN, LOT: D5330150, IP, 150 mg / kg, administered 10 minutes before administration to each group). The mice were anesthetized and then placed in the IVIS® Lumina LT Series III small animal bio-optical imaging system, in the order of tail marks 1 to 5. Whole-body fluorescence was detected to evaluate the delivery and expression of the prepared luciferase mRNA.

[0420] At the end of the study, the mice were sacrificed by cervical dislocation, and the liver, spleen, stomach, and intestines were collected. The organs were removed according to the corresponding positions of 1# to 5#, and the fluorescence of the organs was detected.

[0421] As shown in Figures 2A to 2D, the groups of Compounds 2A, 5A, and 13 of the present invention achieved significant extrahepatic delivery effects, which were significantly better than those of the groups of MC3 and Compound 20A, indicating that the compounds of the present invention reduced the toxicity of liver accumulation and improved the extrahepatic delivery properties.

[0422] Experimental Example 5: Toxicity Assay The animals were divided into groups (5 animals per group) based on body weight on the day before administration (Zhejiang Vital River Laboratory Animal Technology Co., Ltd., male, body weight: 170-220 g). The animals' body weights were measured before administration, and lipid nanoparticle compositions based on Compound 2A, Compound 5A, Compound 13, or Compound 20A (prepared according to Test Example 1) of the present invention were intravenously administered in a single dose based on body weight. Luciferase mRNA was encapsulated in the lipid nanoparticle compositions. SM102 LNP, MC3 LNP (each prepared according to the method and ratios in Test Example 1), and phosphate-buffered saline (PBS) were set up in parallel as control groups.

[0423] The administration volume is 10 mL / kg and the dose is 3 mg / kg.

[0424] Detection frequency: Non-anticoagulated blood samples were collected before and 24 hours after administration. The blood collection method was as follows: Non-anticoagulated blood was collected (the mice were fasted overnight before collection but had free access to water and were fed after collection). The non-anticoagulated whole blood was left at room temperature for 30 minutes and then centrifuged at 5000 rpm for 5 minutes. Serum (200 μL) was aspirated, and serum ALT (alanine aminotransferase) and AST (aspartate aminotransferase) were measured using an automated biochemical detector (Automated Bio-Chemical Detector 7060 Hitachi).

[0425] During the study, four rats in the MC3 group died, and all five rats in the Compound 20A group died.

[0426] The results shown in Figures 3A and 3B indicate that the aspartate aminotransferase and alanine aminotransferase values ​​of Compounds 2A, 5A, and 13 of the present invention are lower than those of the pH-responsive lipids MC3 and SM102 used in LNP in commercially available products, respectively, indicating that the compounds of the present invention have low hepatotoxicity and high biological safety.

[0427] Although the present application has been described above in relation to the embodiments thereof, the present application should not be limited by the specific embodiments and fields of application described above. The specific embodiments described above are intended to illustrate, but not limit, the scope of the present disclosure. A person skilled in the art can create multiple forms within the scope of the claims of the present application under the concept of the specification of the present application without departing from the scope of the claims of the present application.

Claims

1. Formula (I), 【Chemistry 1】 [In the formula, R １ teeth, 【Chemistry 2】 (In the formula, R １ａ and R １ｂ are independently selected from H or C1-C6 alkyl, and R ２ and R ３ are independently selected from H or C1-C6 alkyl, or R １ and R ２ together with the carbon atoms to which they are attached form a substituted or unsubstituted 5- to 7-membered nitrogen-containing heterocycle, R ３ is selected from H or C1-C6 alkyl, or R １ and R ３ together with the carbon atoms to which they are attached form a ring A which is a substituted or unsubstituted 5- to 7-membered carbocyclic or heterocyclic ring; R ２ is selected from H or C1-C6 alkyl; R ４ , R ５ and R ６ are independently selected from C1-C14 alkyl; X and Y are independently selected from O or S; X １ and X ２ is independently selected from C=O or O; Y １ and Y ２ are independently selected from C=O or O, with the proviso that X １ and Y １ are not both C=O or O, and X ２ and Y ２ are not both C=O or O; m and n are independently selected from 0, 1, or 2, with the proviso that m and n cannot both be 0; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and q is selected from 0, 1, 2, 3, 4, 5, or 6, or a salt or stereoisomer thereof.

2. The compound of formula (I) is represented by formula (IA): 【Transformation 3】 2. The compound of claim 1, wherein q is selected from 1, 2, 3, or 4, or a salt or stereoisomer thereof.

3. R ４ is a C9, C10, C11, or C12 straight chain alkyl; R ５ and R ６ 3. The compound of claim 1 or 2, or a salt or stereoisomer thereof, wherein: are each independently selected from a C6, C7, C8, or C9 straight chain alkyl.

4. The compound according to any one of claims 1 to 3, wherein o is 5 or 6, or a salt or stereoisomer thereof.

5. The compound according to any one of claims 1 to 4, or a salt or stereoisomer thereof, wherein p is 7 or 8.

6. 6. The compound according to any one of claims 1 to 5, or a salt or stereoisomer thereof, wherein X and Y are both O.

7. 7. The compound according to any one of claims 1 to 6, wherein n is 1 and m is 0; or n is 1 and m is 1; or n is 0 and m is 1, or a salt or stereoisomer thereof.

8. The compound of formula (I) has the following structure: 【Chemistry 4】 The compound according to any one of claims 1 to 7, or a salt or stereoisomer thereof, selected from:

9. The compound of formula (I) has the following structure: 【Transformation 5】 9. The compound according to any one of claims 1 to 8, or a salt or stereoisomer thereof, selected from:

10. The compound of formula (I) has the following structure: 【Transformation 6】 10. The compound according to any one of claims 1 to 9, or a salt or stereoisomer thereof, selected from:

11. C1-C6 alkyl is —CH ３ , -CH ２ CH ３ or -CH(CH ３ ) ２ 11. The compound of any one of claims 1 to 10, or a salt or stereoisomer thereof, independently selected from:

12. R １ａ and R １ｂ But -CH ３ are independently selected from R ２ and R ３ is selected independently from H; R ４ is a C10, C11 or C12 straight chain alkyl; R ５ and R ６ are the same and are selected from C6, C7, and C8 straight chain alkyl; X and Y are independently selected from O; X １ and X ２ is independently selected from C═O or O; Y １ and Y ２ are independently selected from C=O or O, with the proviso that X １ and Y １ are not both C=O or O, and X ２ and Y ２ are not both C=O or O; m is 0 and n is 1; o is 5 or 6; p is 7 or 8, for example, p is 7; 12. A compound according to any one of claims 1 to 11, or a salt or stereoisomer thereof, wherein q is 2, 3 or 4, for example q is 2 or 3, for example q is 2.

13. X １ and X ２ are both C═O, and Y １ and Y ２ The compound according to any one of claims 1 to 12, or a salt or stereoisomer thereof, wherein

14. X １ is O and Y １ is C=O, and X ２ is C=O, and Y ２ The compound according to any one of claims 1 to 12, or a salt or stereoisomer thereof, wherein is O.

15. X １ is C=O, and Y １ is O and X ２ is O and Y ２ is C=O; or X １ and X ２ are both O and Y １ and Y ２ The compound according to any one of claims 1 to 12, or a salt or stereoisomer thereof, wherein both are C=O.

16. R ４ The compound according to any one of claims 1 to 15, or a salt or stereoisomer thereof, wherein is a C10-C11 straight chain alkyl.

17. R ５ and R ６ The compound according to any one of claims 1 to 16, or a salt or stereoisomer thereof, wherein both are C8 straight chain alkyl.

18. o is 5 and R ４ is a C11 linear alkyl, p is 7, and R ５ and R ６ are both C8 linear alkyl, or o is 6 and R ４ is a C10 linear alkyl, p is 7, and R ５ and R ６ The compound according to any one of claims 1 to 17, or a salt or stereoisomer thereof, wherein both are C8 straight chain alkyl. 【Request Item 19】 【Chemistry 7】 【Transformation 8】 【Chemistry 9】 2. The compound of claim 1, or a salt or stereoisomer thereof, selected from: 【Request Item 20】 【Chemistry 10】 【Chemistry 11】 2. The compound of claim 1, or a salt or stereoisomer thereof, selected from:

21. A process for preparing a compound according to any one of claims 1 to 20, wherein compound (IA) is obtained from compound J as starting material, 【Chemistry 12】 During the ceremony, Z １ is selected from OH, Z ２ and Z ３ are each independently selected from OH or SH; Z ４ is selected from OMs; X, Y, X １ , X ２ , Y １ , Y ２ , R １ａ , R １ｂ , R ２ , R ３ , R ４ , R ５ , R ６ , m, n, o, p and q are as defined in any one of claims 1 to 20; Compound J is reacted with compound K in a benzene-based solution of pyridinium 4-methylbenzenesulfonate to provide compound L, and the benzene-based solvent used in the benzene-based solution is preferably toluene; The compound L is (CH ３ SO ２ ) ２ O in a haloalkane solution to provide compound M, said haloalkane being preferably DCM, dichloroethane or chloroform; The method wherein said compound M is reacted in a polar aprotic solvent to form compound IA, said polar aprotic solvent preferably being THF.

22. Compound J is obtained from compound H and compound C as starting materials, 【Chemistry 13】 wherein X is selected from halogen; X １ , X ２ , R ４ , R ５ , R ６ , o and p are as defined in any one of claims 1 to 20; The compound H is a carbonate, NBu ４ and a polar aprotic solvent to provide compound I, wherein the carbonate is preferably Cs ２ CO ３ or K ２ CO ３ wherein the polar aprotic solvent is preferably DMF or DMA; 22. The preparation method according to claim 21, wherein compound I reacts at pH 1-5 to provide compound J, preferably the pH is adjusted by hydrochloric acid solution, and the reaction solvent is a haloalkane, preferably DCM, dichloroethane or chloroform.

23. Compound F reacts with compound G to provide compound H; 【Chemistry 14】 wherein X is selected from halogen; X １ , Y １ , R ４ and o are as defined in any one of claims 1 to 20; The compound F is a carbonate, NBu ４ with compound G in the presence of I and a polar aprotic solvent to provide compound H, wherein the carbonate is preferably Cs ２ CO ３ or K ２ CO ３ 23. The process according to claim 22, wherein the polar aprotic solvent is preferably DMF or DMA.

24. 21. A nanoparticle composition comprising a lipid component comprising a compound according to any one of claims 1 to 20; preferably, said nanoparticle composition is a lipid nanoparticle (LNP).

25. the lipid component further comprises a phospholipid, a structured lipid, and / or a PEG lipid; The phospholipids preferably contain the following compounds: Dilauryl lecithin (DLPC), Dimyristoylphosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dioleoylphosphatidylcholine (DUPC), Palmitoyloleoylphosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 lysoPC), 1,2-divinyl-sn-glycero-3-phosphocholine, 1,2-diarylacyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphorylethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-diethenol-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexaenoate-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphate-(1-glycerol) sodium salt (DOPG) or sphingomyelin; For example, the phospholipid is DOPE or DSPC; the structured lipid is preferably selected from one or more of cholesterol, coprosterol, sitosterol, ergosterol and stigmasterol; for example, the structured lipid is cholesterol; and / or 25. The nanoparticle composition of claim 24, wherein the PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, or PEG-modified dialkylglycerol.

26. 26. The nanoparticle composition of claim 24 or 25, wherein the lipid component further comprises a cationic lipid and / or a pH-responsive lipid.

27. further comprising a therapeutic and / or prophylactic agent, wherein the therapeutic and / or prophylactic agent is selected from a vaccine, or a compound capable of inducing an immune response, and / or a nucleic acid; The nanoparticle composition of any one of claims 24 to 26, wherein the nucleic acid is preferably RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

28. the encapsulation efficiency of the therapeutic agent and / or the prophylactic agent is 50% or more; or 80% or more; or 90% or more; and / or the nanoparticle composition has an average particle size of 50 nm to 110 nm; and / or The nanoparticle composition of any one of claims 24 to 27, wherein the nanoparticle composition has a polydispersity index of 0.04 to 0.

20.

29. 21. Use of a compound according to any one of claims 1 to 20, or a salt or stereoisomer thereof, in the preparation of a lipid nanoparticle composition.

30. A pharmaceutical composition comprising the nanoparticle composition of any one of claims 24 to 28 and a pharmaceutically acceptable excipient.

31. A method for delivering a therapeutic and / or prophylactic agent to a mammalian cell, comprising administering to a subject the nanoparticle composition of any one of claims 24 to 28 or the pharmaceutical composition of claim 30, wherein the administering comprises contacting the cell with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and / or prophylactic agent to the cell; e.g., the mammalian cell is in a mammal; e.g., the mammal is a human; e.g., the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

32. A method for producing a polypeptide of interest in a mammalian cell, comprising contacting the cell with the nanoparticle composition of any one of claims 24 to 28 or the pharmaceutical composition of claim 30 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, and the mRNA can be translated in the cell to produce the polypeptide of interest; e.g., the mammalian cell is in a mammal; e.g., the mammal is a human; e.g., the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

33. A method for treating or preventing a disease or condition in a mammal, such as a human, comprising administering to the mammal a therapeutically or prophylactically effective amount of the nanoparticle composition of any one of claims 24 to 28, or the pharmaceutical composition of claim 30; for example, the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity; for example, the disease or condition is selected from infectious diseases, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases, or metabolic diseases; for example, the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

34. A method for specifically delivering a therapeutic and / or prophylactic agent to an organ of a mammal, comprising administering to the mammal a nanoparticle composition described in any one of claims 24 to 28 or a pharmaceutical composition described in claim 30, wherein the administering comprises contacting the organ of the mammal with the nanoparticle composition or the pharmaceutical composition, thereby delivering the therapeutic and / or prophylactic agent to the organ, e.g., wherein the mammal is a human; e.g., the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation; e.g., the mammal is pretreated 24 hours or less before the contacting or administering step; e.g., about 1 hour before the contacting or administering step.

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