CN117946201A - Lipid compounds and lipid nanoparticle compositions - Google Patents

Abstract

The application relates to a lipid compound and a lipid nanoparticle composition, in particular to a lipid compound, the structural formula of which is shown in a formula (I), and salts and isomers thereof, and a nanoparticle composition containing the compound or the salts or the isomers thereof.

Description

Lipid compounds and lipid nanoparticle compositions

Technical Field

The present invention relates to the field of biotechnology, and in particular to lipid compounds and lipid nanoparticle compositions.

Background Art

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

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

LNPs generally include one or more cationic lipids and/or amino (ionizable) lipids, phospholipids containing polyunsaturated lipids, structural lipids (such as sterols) and/or lipids containing polyethylene glycol (PEG lipids). Cationic and/or ionizable lipids include, for example, amine-containing lipids that can be easily protonated.

Lipid nanoparticle (LNP) formulations represent a revolution in the field of nucleic acid delivery. Onpattro ^TM is an early example of a lipid nanoparticle product approved for clinical use. Onpattro ^TM is a lipid nanoparticle-based short interfering RNA (siRNA) drug 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 leading LNP-based COVID-19 mRNA vaccine.

Onpattro ^TM LNP formulation consists of four major lipid components, namely: ionizable amino lipid MC3, distearoylphosphatidylcholine (DSPC), cholesterol and polyethylene glycol-conjugated lipid (PEG-lipid), with molar ratios of 50/10/38.5/1.5 respectively. Onpattro ^TM is still considered the gold standard for LNP research comparison.

Although great progress has been made in the research related to LNP-mediated nucleic acid delivery, it is known that Onpattro ^TM formulations accumulate primarily in liver tissue. The ability of LNPs to accumulate in organs and tissues other than the liver will greatly expand the clinical use of these delivery systems.

Summary of the invention

Based on this, the present application discloses a lipid compound and a lipid nanoparticle composition comprising the compound, wherein the lipid has the advantages of high encapsulation rate, high expression, spleen targeting, etc., and also has the characteristics of low toxicity, especially low liver toxicity.

Specifically, this application adopts the following technical solutions

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

_X1 and _X2 are independently selected from C=O or O, _Y1 and _Y2 are independently selected from C=O or O, provided that _X1 and _Y1 , _X2 and _Y2 are not C=O or O at the same time; _R1 is R; _R2 and _R3 are independently selected from H, C1-C14 alkyl, R, provided that _R2 and _R3 are not H at the same time;

The R is

Ra, Rb, Rc, Rd, Re are independently selected from H, C1-C6 alkyl,

R' is selected from C1-C10 alkyl, C1-C10 alkenyl;

R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl;

Dashed lines represent single or double bonds;

n is selected from 0, 1, 2, 3, 4, 5, 6;

o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

2. The compound according to item 1, wherein R ₂ and R ₃ are independently selected from H and C1-C14 alkyl, provided that R ₂ and R ₃ are not H at the same time.

3. The compound according to item 2, wherein R ₂ and R ₃ are C6-C11 straight-chain or branched alkyl groups.

4. The compound according to item 3, wherein R ₂ and R ₃ are C8 straight-chain or branched alkyl groups.

5. The compound according to item 2, wherein _R2 is H, and _R3 is a C10-C12 straight-chain or branched alkyl group.

6. The compound according to item 5, wherein _R2 is H, and _R3 is a C11 straight-chain or branched alkyl group.

7. The compound according to item 1, wherein _R2 is H and _R3 is R.

8. The compound according to item 1-7, wherein R' is a C8 alkyl group or a C8 alkenyl group.

9. The compound according to item 8, wherein R' is

Dashed lines represent single or double bonds.

10. The compound according to any one of items 1 to 9, wherein R is: Best

11. The compound according to any one of items 1-10, wherein o is selected from 1, 2, 3, 4, 5, 6, and 7.

12. The compound according to item 11, wherein o is 5.

13. The compound according to any one of items 1-12, wherein p is selected from 1, 2, 3, 4, 5, 6, 7.

14. The compound according to item 13, wherein p is 7.

15. A compound according to any one of items 1-14, wherein n is 2.

16. A compound according to any one of items 1-14, wherein n is 6.

17. The compound according to any one of items 1 to 16, wherein X ₁ and X ₂ are C═O, and Y ₁ and Y ₂ are O.

18. The compound according to any one of items 1 to 16, wherein X ₁ is O, Y ₁ is C═O, X ₂ is C═O, and Y ₂ is O.

19. The compound according to any one of items 1 to 16, wherein X ₁ is C═O, Y ₁ is O, X ₂ is O, and Y ₂ is C═O.

20. The compound according to any one of items 1 to 16, wherein X ₁ and X ₂ are O, and Y ₁ and Y ₂ are C═O.

21. A compound according to any one of items 1 to 20, which is a compound of formula (II):

22. A compound or a salt thereof or an isomer thereof, wherein the compound is selected from

Compound 1

Compound 2

Compound 3

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Compound 9

Compound 10

Compound 11

Compound 12

Compound 13

Compound 14

Compound 15

Compound 16

23. The compound according to item 22, or a salt thereof, or an isomer thereof, wherein the compound is selected from

Compound 1

Compound 2

24. A nanoparticle composition comprising a lipid component, wherein the lipid component comprises a compound as described in any one of items 1-23.

25. A nanoparticle composition according to claim 24, wherein the lipid component further comprises a phospholipid.

26. The nanoparticle composition according to claim 25, wherein the phospholipid is selected from one or more of the following compounds: dilauroyl phosphatidylcholine (DLPC), dimyristoyl phosphatidylcholine (DMPC), dioleoyl phosphatidylcholine (DOPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DUPC), palmitoyl oleoyl phosphatidylcholine (POPC), 1,2-di-O-octadecyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesteryl dimethylsuccinate-sn-glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-divinyl-sn-glycero-3-phosphocholine, 1,2-diaryl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE), 1,2-dihydroxytin-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-divinyl-sn-glycero-3-phosphoethanolamine, 1,2-diaryl-sn-glycero-3-phosphoethanolamine, 1,2-dithiohexaenoic acid-sn-glycero-3-phosphoethanolamine, 1,2-diol-sn-glycero-3-phospho-(1-glycerol) sodium salt (DOPG) or sphingomyelin.

27. A nanoparticle composition according to claim 25, wherein the phospholipid is DOPE.

28. A nanoparticle composition according to claim 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 structural lipid.

30. The nanoparticle composition of claim 29, wherein the structural lipid is selected from one or more of cholesterol, coprostanol, sitosterol, ergosterol, and stigmasterol.

31. A nanoparticle composition according to item 29, wherein the structural lipid is cholesterol.

32. The nanoparticle composition of any one of items 24 to 31, wherein the lipid component further comprises a PEG lipid.

33. A nanoparticle composition according to any one of claim 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 of any one of items 24 to 33, wherein the lipid component further comprises cationic and/or ionizable lipids.

35. The nanoparticle composition according to any one of items 24 to 34 further includes a therapeutic agent and/or a preventive agent, wherein the therapeutic agent and/or the preventive agent is selected from a vaccine or a compound capable of inducing an immune response, or a nucleic acid, preferably the nucleic acid is RNA, and the RNA is selected from one or more of siRNA, aiRNA, miRNA, dsRNA, shRNA or mRNA.

36. The nanoparticle composition of any one of items 24 to 35, wherein the encapsulation efficiency of the therapeutic agent and/or prophylactic agent is ≥50%; or ≥80%; or ≥90%.

37. The nanoparticle composition according to any one of items 24 to 36, wherein the average particle size of the nanoparticle composition is 50 nm to 110 nm.

38. The nanoparticle composition according to any one of items 24 to 37, wherein the dispersibility index of the nanoparticle composition is 0.006-0.20.

39. Use of a compound according to any one of items 1 to 23 in the preparation of a lipid nanoparticle composition.

40. A pharmaceutical composition comprising the nanoparticle composition of any one of items 24 to 38 and a pharmaceutically acceptable carrier.

41. A method for delivering a therapeutic and/or prophylactic agent to mammalian cells, the method comprising administering to a subject 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 cells with the nanoparticle composition or the pharmaceutical composition to deliver the therapeutic and/or prophylactic agent to the cells.

42. A method according to claim 41, wherein the mammalian cell is in a mammal.

43. The method of item 41 or 42, wherein the mammal is a human.

44. The method of any one of items 41 to 43, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

45. A method for producing a target polypeptide in a mammalian cell, the method comprising contacting the cell with a nanoparticle composition described in any one of items 24 to 38 or a pharmaceutical composition described in item 40 to deliver a therapeutic and/or prophylactic agent to the cell, wherein the therapeutic and/or prophylactic agent is mRNA, which encodes the target polypeptide, such that the mRNA can be translated in the cell to produce the target polypeptide.

46. A method according to claim 45, wherein the mammalian cell is in a mammal.

47. A method according to any one of items 45 or 46, wherein the mammal is a human.

48. The method of any one of items 45 to 47, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

49. A method for treating a disease or disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition of any one of items 24 to 38 or the pharmaceutical composition of item 40.

50. A method according to claim 49, wherein the disease or condition is characterized by dysfunctional or abnormal protein or polypeptide activity.

51. A method according to claim 49 or 50, wherein the disease or condition is selected from an infectious disease, cancer and proliferative diseases, genetic diseases, autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular diseases, renal vascular diseases or metabolic diseases.

52. The method according to any one of items 49 to 51, wherein the mammal is a human.

53. The method of any one of items 49 to 52, wherein the nanoparticle composition or the pharmaceutical composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

54. A method for specifically delivering a therapeutic and/or prophylactic agent to a mammalian organ, the method comprising administering to the mammal a nanoparticle composition described in any one of items 24 to 38 or a pharmaceutical composition described in item 40, wherein the administration comprises contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and/or prophylactic agent to the organ.

55. The method of claim 54, wherein the mammal is a human.

56. The method of item 54 or 55, wherein the nanoparticle composition is administered intravenously, intramuscularly, intradermally, subcutaneously, intranasally, or by inhalation.

57. The method of any one of items 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 items 54 to 57, wherein the mammal is pretreated about one hour prior to the contacting or administering step.

59. Use of the nanoparticle composition described in any one of items 24 to 38 or the pharmaceutical composition described in item 40 in the preparation of a medicament for treating a disease or condition in a mammal.

60. Use of the nanoparticle composition of any one of items 24 to 38 or the pharmaceutical composition of item 40 in treating a disease or condition in a mammal.

The compound of formula (I) provided in the present application, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof,

X ₁ and X ₂ are independently selected from C═O or O, and Y ₁ and Y ₂ are independently selected from C═O or O, provided that X ₁ and Y ₁ , X ₂ and Y ₂ are not C═O or O at the same time;

_R1 is R,

R ₂ and R ₃ are independently selected from H, C1-C14 alkyl, and R, provided that R ₂ and R ₃ are not H at the same time;

The R is

Ra, Rb, Rc, Rd, Re are independently selected from H, C1-C6 alkyl,

R' is selected from C1-C10 alkyl, C1-C10 alkenyl,

Dashed lines represent single or double bonds;

R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl;

n is selected from 0, 1, 2, 3, 4, 5, 6;

o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

In some embodiments, R ₂ and R ₃ are independently selected from H, C1-C14 alkyl, with the proviso that R ₂ and R ₃ are not H at the same time.

In some embodiments, R ₂ and R ₃ are C6-C11 straight chain or branched chain alkyl groups.

In some embodiments, R ₂ and R ₃ are C8 straight or branched chain alkyl groups.

In some embodiments, R ₂ is H, and R ₃ is a C10-C12 straight chain or branched alkyl group.

In some embodiments, R ₂ is H, and R ₃ is a C11 straight or branched chain alkyl group.

In some embodiments, _R2 is H and _R3 is R.

In some embodiments, R' is a C8 alkyl or C8 alkenyl.

In some embodiments, R' is Dashed lines represent single or double bonds.

In some embodiments, R is: Best

In some embodiments, o is selected from 1, 2, 3, 4, 5, 6, 7.

In some embodiments, o is 5.

In some embodiments, p is selected from 1, 2, 3, 4, 5, 6, 7.

In some embodiments, p is 7.

In some embodiments, n is 2.

In some embodiments, n is 6.

In some embodiments, X ₁ and X ₂ are C═O, and Y ₁ and Y ₂ are O.

In some embodiments, X ₁ is O, Y ₁ is C═O, X ₂ is C═O, and Y ₂ is O.

In some embodiments, X ₁ is C=O, Y ₁ is O, X ₂ is O, and Y ₂ is C=O.

In some embodiments, X ₁ and X ₂ are O, and Y ₁ and Y ₂ are C═O.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl groups; X ₁ and X ₂ are independently selected from C=O or O, and Y ₁ and Y ₂ are independently selected from C=O or O, with the proviso that X ₁ and Y ₁ , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R _{7 are independently selected from H, C1-C6 alkyl groups; the dotted line represents a single bond or a double bond; n is selected from 0, 1, 2, 3, 4, 5, 6} ; the p is selected from 1, 2, 3, 4, 5, 6, 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are 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 , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is selected from 0, 1, 2, 3, 4, 5, 6; the p is selected from 1, 2, 3, 4, 5, 6, 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl groups; X ₁ and X ₂ are independently selected from C=O or O, and Y ₁ and Y ₂ are independently selected from C=O or O, with the proviso that X ₁ and Y ₁ , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl groups; the dotted line represents a single bond or a double bond; n is selected from 0, 1, 2, 3, 4, 5, 6; the p is 7; and the o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are 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 , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is selected from 0, 1, 2, 3, 4, 5, 6; the p is 7; the o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl groups; X ₁ and X ₂ are independently selected from C=O or O, and Y ₁ and Y ₂ are independently selected from C=O or O, with the proviso that X ₁ and Y ₁ , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl groups; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are 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 , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl groups; X ₁ and X ₂ are independently selected from C=O or O, and Y ₁ and Y ₂ are independently selected from C=O or O, with the proviso that X ₁ and Y ₁ , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl groups; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are 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 , X ₂ and Y ₂ are not C=O or O at the same time; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl; X ₁ and X ₂ are C═O, Y ₁ and Y ₂ are O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are C═O, Y ₁ and Y ₂ are O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl; X ₁ and X ₂ are C═O, Y ₁ and Y ₂ are O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are C═O, Y ₁ and Y ₂ are O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight chain or branched alkyl; X ₁ is O, X ₂ is C=O, Y ₁ is C=O, Y ₂ is O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ is O, X ₂ is C=O, Y ₁ is C=O, Y ₂ is O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight chain or branched alkyl; X ₁ is O, X ₂ is C＝O, Y ₁ is C＝O, Y ₂ is O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ is O, X ₂ is C=O, Y ₁ is C=O, Y ₂ is O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight chain or branched alkyl; X ₁ is C＝O, X ₂ is O, Y ₁ is O, Y ₂ is C＝O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ is C=O, X ₂ is O, Y ₁ is O, Y ₂ is C=O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight chain or branched alkyl; X ₁ is C＝O, X ₂ is O, Y ₁ is O, Y ₂ is C＝O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ is C=O, X ₂ is O, Y ₁ is O, Y ₂ is C=O; R ₁ is R; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl; X ₁ and X ₂ are O, Y ₁ and Y ₂ are C═O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are O, Y ₁ and Y ₂ are C═O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 2; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C6-C11 straight or branched alkyl; X ₁ and X ₂ are O, Y ₁ and Y ₂ are C═O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, R ₂ and R ₃ are C8 straight or branched alkyl; X ₁ and X ₂ are O, Y ₁ and Y ₂ are C═O; R ₁ is R; R ₄ , R ₅ , R ₆ and R ₇ are independently selected from H and C1-C6 alkyl; the dotted line represents a single bond or a double bond; n is 6; p is 7; and o is 5.

In some embodiments, it is a compound of formula (II) or a pharmaceutically acceptable salt or a stereoisomer thereof:

In some embodiments, it is a compound of formula (II) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is selected from 2, 3, 4, 5, 6; o is selected from 3, 4, 5; p is selected from 7 or 8; R1 is R2 and R3 are each independently selected from C7-C9 straight chain alkyl.

In some embodiments, it is a compound of formula (II) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is selected from 2, 3, 4, 5, 6; o is 5; p is 7; R1 is R2 and R3 are each independently selected from C7-C9 straight chain alkyl.

In some embodiments, it is a compound of formula (II) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is selected from 2, 3, 4, 5, 6; o is 5; p is 7; R1 is R2 and R3 are each independently a C8 straight chain alkyl group.

In some embodiments, it is a compound of formula (II) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein n is selected from 2 or 6; o is 5; p is 7; R1 is R2 and R3 are each independently selected from C8 straight chain alkyl.

In some embodiments, the compound is selected from

Compound 1

Compound 2

Compound 3

Compound 4

Compound 5

Compound 6

Compound 7

Compound 8

Compound 9

Compound 10

Compound 11

Compound 12

Compound 13

Compound 14

Compound 15

Compound 16

In some embodiments, the compound is selected from

Compound 1

Compound 2

In some embodiments, Compound 1 and its stereoisomers are selected from

Compound 1a or

Compound 1b or a racemate of Compound 1a and Compound 1b.

In some embodiments, Compound 2 and its stereoisomers are selected from

Compound 2a or

Compound 2b or a racemate of Compound 2a and Compound 2b.

Effects of the Invention

The compounds of the present application can be used for the preparation of lipid nanoparticles. The nanoparticle compositions containing the compounds provided in the present application can achieve the encapsulation and delivery of therapeutic/preventive agents, safely deliver the therapeutic/preventive agents to the targeted location, achieve high expression, exert the effects of the therapeutic/preventive agents, and at the same time have the characteristics of low toxicity, especially low liver toxicity.

The lipid nanoparticles prepared in the present application have a small average particle size and a high encapsulation efficiency, and have broad application prospects in the field of drug delivery.

BRIEF DESCRIPTION OF THE DRAWINGS

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

FIG1 shows the luciferase fluorescence intensity of different LNP formulations 6 hours after intravenous injection.

FIG2 shows the luciferase fluorescence intensity of different LNP formulations in the spleen 24 hours after intravenous injection.

FIG3 shows the luciferase fluorescence intensity of different LNP formulations 6 hours after intramuscular injection.

DETAILED DESCRIPTION

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

Terms and Definitions

As used herein, the term "alkyl" refers to a group comprising one or more carbon atoms (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty or more carbon atoms), which is optionally substituted. For example, the term "C1-C14 alkyl" refers to an optionally substituted straight or branched saturated hydrocarbon including 1-14 carbon atoms, the term "C1-C10 alkyl" refers to an optionally substituted straight or branched saturated hydrocarbon including 1-10 carbon atoms, and the term "C1-C6 alkyl" refers to an optionally substituted straight or branched saturated hydrocarbon including 1-6 carbon atoms. Unless otherwise indicated, the alkyl described herein refers to unsubstituted and substituted alkyl.

As used herein, the term "alkenyl" refers to one or more carbon atoms (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty or more carbon atoms), which is optionally substituted. Such as the term "C1-C14 alkenyl" refers to an optionally substituted straight or branched unsaturated hydrocarbon including 1-14 carbon atoms, the term "C1-C10 alkenyl" refers to an optionally substituted straight or branched unsaturated hydrocarbon including 1-10 carbon atoms, and the term "C1-C6 alkenyl" refers to an optionally substituted straight or branched unsaturated hydrocarbon including 1-6 carbon atoms. Unless otherwise indicated, alkenyl described herein refers to unsubstituted and substituted alkenyl.

Unless otherwise specified, alkyl groups may be optionally substituted. The optional substituents may be selected from, but are not limited to, 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), carbonyls (e.g., -C(O)R, or represented by C=O), acyl halides (e.g., -C(O)X, wherein X is a halide selected from bromide, fluoride, chloride and iodide), carbonates (e.g., -OC(O)OR), alkoxy groups (e.g., -OR), acetals, phosphates, thiols (e.g., -SH), sulfoxides (e.g., -S(O)R), sulfites (e.g., -S(O)OH), sulfonic acids (e.g., -S(O) _2OH ), thiols (e.g., -C(S)H), sulfates, sulfonyl groups (e.g., -S(O) _2- ), amides (e.g., -C(O)NR ₂ or -N(R)C(O)R), azido (e.g. -N ₃ ), nitro (e.g. -NO ₂ ), cyano (e.g. -CN), isocyano (e.g. -NC), acyloxy (e.g. -OC(O)R), amino (e.g. -NR ₂ , -NRH or -NH ₂ ), carbamoyl (e.g. -OC(O)NR ₂ , -OC(O)NRH or -OC(O)NH ₂ ), sulfonamide, alkyl, alkenyl and cyclic (e.g. carbocyclyl or heterocyclyl). In any of the foregoing, R is alkyl or alkenyl as defined herein. In some embodiments, the substituents themselves can be further substituted with one, two, three, four, five or six substituents, e.g., as defined herein. For example, C _1-6 alkyl can be further substituted with 1, 2, 3, 4, 5 or 6 substituents as described herein.

As used herein, the term "compound" is meant to include all isomers and isotopes of the described structure. "Isotopes" refer to atoms with the same atomic number, but with different mass numbers due to different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium. In addition, the compounds, salts or complexes of the present application can be prepared by conventional methods in combination with solvents or water molecules to form sols and hydrates.

As used herein, the term "contacting" 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. The method of contacting a cell with an external entity in vivo and in vitro is well known in the biological field. For example, a nanoparticle composition can be contacted with a mammalian cell placed in a mammal by a variety of routes of administration (e.g., intravenous, intramuscular, intradermal and subcutaneous), and can involve a variety of nanoparticle compositions. In addition, the nanoparticle composition can contact more than one mammalian cell.

As used herein, the term "delivery" refers to providing an entity to a destination. For example, delivering a therapeutic agent and/or a prophylactic agent to a subject may include administering to the subject a nanoparticle composition comprising a therapeutic agent and/or a prophylactic agent (e.g., by intravenous, intramuscular, intradermal, or subcutaneous route). Administering a nanoparticle composition to a mammal or mammalian cell may involve contacting one or more cells with the nanoparticle composition.

As used herein, the term "enhanced delivery" refers to the delivery of more (e.g., at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times) therapeutic and/or preventive drugs to target tissues (e.g., mammal liver) by nanoparticles compared to the level of therapeutic and/or preventive drugs delivered to target tissues (e.g., MC3, KC2 or DLinDMA) by control nanoparticles. Can be by comparing the amount of protein produced in tissue with the weight of the tissue, the therapeutic and/or preventive amount in tissue with the weight of tissue, the amount of protein produced in tissue with the amount of total protein in tissue, or the amount of treatment and/or preventive agent in tissue with the total treatment and/or preventive agent in the tissue. It should be understood that the enhanced delivery of nanoparticles to target tissues does not need to be determined in the treated subject, but can be determined in a surrogate such as an animal model (e.g., rat model). In some embodiments, nanoparticle compositions comprising a compound of Formula (I) or (II) have substantially the same level of delivery enhancement regardless of the route of administration.

As used herein, the term "specific delivery" or "specific transport" refers to the delivery of more (e.g., at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times) therapeutic and/or preventive drugs to a target tissue (e.g., mammalian liver) by nanoparticles than to a non-target tissue. The level of delivery of nanoparticles to a specific tissue can be measured by comparing the weight of protein produced in a tissue with the weight of the tissue, comparing the therapeutic and/or preventive amount in a tissue with the weight of the tissue, comparing the protein weight produced in a tissue with the total protein weight in the tissue, or comparing the therapeutic and/or preventive amount in a tissue with the total therapeutic and/or preventive amount in the tissue.

As used herein, "encapsulation efficiency" refers to the amount of therapeutic and/or prophylactic agent that becomes part of a nanoparticle composition, relative to the total amount of therapeutic or prophylactic agent used in preparing the nanoparticle composition. For example, if 97 mg of the therapeutic and/or prophylactic agent is 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, enclosure, surrounding or encapsulation.

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

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

As used herein, the term "in vivo" refers to events that occur within an organism (eg, an animal, plant or microorganism, or a cell or tissue thereof).

As used herein, the term "ex vivo" refers to an event that occurs outside an organism (e.g., an animal, plant, or microorganism, or a cell or tissue thereof). An ex vivo event can occur in an environment that is minimally altered from the natural (e.g., in vivo) environment.

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

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/ionizable lipids, PEGylated lipids, structural lipids or other lipids, such as phospholipids.

As used herein, a "linker" is a portion connecting two parts, for example, the connection between two nucleosides of a cap species. A connector can include one or more groups, including but not limited to a phosphate group (e.g., phosphate, borophosphate, thiophosphate, selenophosphate, and phosphate), an alkyl, an amide, or a glycerol. For example, two nucleosides of a cap analog can be connected at their 5' positions by a triphosphate group or by a chain comprising two phosphate moieties and a boric acid phosphate moiety.

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

As used herein, "modified" refers to non-natural. For example, RNA can be a modified RNA. That is, the RNA can include one or more non-naturally occurring nucleobases, nucleosides, nucleotides or linkers. A "modified" substance may also be referred to herein as an "engineered" substance. The substance may be modified or engineered chemically, structurally or functionally. For example, a modified nucleobase species may include one or more non-naturally occurring substitutions.

As used herein, a "nanoparticle composition" is a composition comprising one or more lipids. The particle size of the nanoparticle composition is generally in the order of microns or less, and may include a lipid bilayer. The nanoparticle composition includes lipid nanoparticles (LNP), liposomes (e.g., lipid vesicles), and lipid complexes. For example, the nanoparticle composition may be a liposome having a lipid bilayer having a diameter of 500 nm or less.

As used herein, "naturally occurring" means occurring in nature without human assistance.

As used herein, "patient" refers to a subject who may seek or need treatment, is in need of treatment, is receiving treatment, is about to receive treatment, or is being cared for by a trained professional for a particular disease.

As used herein, "PEG lipid" or "PEGylated lipid" refers to a lipid that comprises a polyethylene glycol component.

The term "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The nanoparticle compositions of the present application may also include salts of one or more compounds. The salt may be a pharmaceutically acceptable salt. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is altered by converting an existing acid or base moiety into its salt form (e.g., by reacting a 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; alkali or organic salts of acidic residues such as carboxylic acids, and the like. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glucoheptonate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, palmitate, pectinate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, tosylate, undecanoate, valerate, and the like.

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

As used herein, "phospholipid" is a lipid comprising a phosphate moiety and one or more carbon chains, such as unsaturated fatty acid chains. Phospholipids may comprise one or more multiple (e.g., double or triple bonds) bonds (e.g., one or more unsaturated bonds). Specific phospholipids may promote fusion with membranes. For example, cationic phospholipids may interact with one or more negatively charged phospholipids of membranes (e.g., cell membranes or intracellular membranes). The fusion of phospholipids with membranes may allow one or more elements containing lipid compositions to pass through the membrane, thereby allowing, for example, one or more elements to be delivered to cells.

As used herein, "polydispersity index" is a ratio that describes the uniformity of the particle size distribution of a system. Small values, such as less than 0.3, indicate a narrow particle size distribution.

As used herein, the term "polypeptide" or "polypeptide of interest" refers to a polymer of amino acid residues, typically linked by peptide bonds, which may be produced naturally (eg, isolated or purified) or synthetically.

As used herein, "RNA" refers to ribonucleic acid that can be naturally or non-naturally occurring. For example, RNA can include modified and/or non-naturally occurring components, such as one or more nucleobases, nucleosides, nucleotides or linkers. RNA can include a cap structure, a chain terminating nucleoside, a stem loop, a polyA sequence and/or a polyadenylation signal. RNA can have a nucleotide sequence encoding a polypeptide of interest. For example, RNA can be a messenger RNA (mRNA). Translation of an mRNA encoding a specific polypeptide, for example, in vivo translation of an mRNA inside a mammalian cell, can produce the encoded polypeptide. RNA can be selected from a non-limiting group including small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), micro RNA (miRNA), double-stranded RNA (dsRNA), small hairpin RNA (shRNA), mRNA and mixtures thereof.

As used herein, a "single unit dose" is a dose of any therapeutic agent that is administered in one dose/at one time/through a single route/at a single point of contact, ie, a single time.

As used herein, a "split dose" is the division of a single unit dose or total daily dose into two or more doses.

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

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.

As used herein, the term "subject" or "patient" refers to any organism to which the composition according to the present application can be administered, e.g., for experimental, diagnostic, preventive and/or therapeutic purposes. Typical subjects include animals (e.g., mammals, e.g., mice, rats, rabbits, non-human primates and humans) and/or plants.

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

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

The term "therapeutic agent" or "prophylactic agent" refers to any agent that has a therapeutic, diagnostic and/or prophylactic effect and/or causes a desired biological and/or pharmacological effect when administered to a subject. Therapeutic agents are also referred to as "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.

As used herein, the term "therapeutically effective amount" refers to an amount of an agent to be delivered (e.g., a nucleic acid, a drug, a composition, a therapeutic agent, a diagnostic agent, a prophylactic agent, etc.) that is sufficient when administered to a subject suffering from or susceptible to an infection, disease, disorder and/or condition to treat, ameliorate the symptoms of, diagnose, prevent and/or delay the onset of the infection, disease, disorder and/or condition.

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

As used herein, the term "treat" refers to partially or completely alleviating, relieving, ameliorating, remitting, delaying the onset of, inhibiting the progression of, reducing the severity of, and/or reducing the incidence of one or more symptoms or features of a particular infection, disease, disorder, and/or condition. For example, "treating" cancer may refer to inhibiting the survival, growth, and/or spread of a tumor. Treatment may be performed on subjects who do not exhibit the disease, disorder, and/or condition and/or on subjects who only exhibit early signs of the disease, disorder, and/or condition in order to reduce the risk of developing a pathological condition associated with the disease, disorder, and/or condition.

In a preferred embodiment, in the compounds of the present invention, R ₂ is selected from C1-C14 alkyl, for example, R ₂ is selected from C2-C14 alkyl, R ₂ is selected from C3-C14 alkyl, R ₂ is selected from C4-C14 alkyl, R ₂ is selected from C5-C14 alkyl, R ₂ is selected from C6-C14 alkyl, R ₂ is selected from C7-C14 alkyl, R ₂ is selected from C8-C14 alkyl, R ₂ is selected from C9-C14 alkyl, R ₂ is selected from C10-C14 alkyl, C10-C13 alkyl, C10-C12 alkyl, C10-C11 alkyl, for example, R ₂ is C1 alkyl, R ₂ is C2 alkyl, R ₂ is C3 alkyl, R ₂ is C4 alkyl, R ₂ is C5 alkyl, R ₂ is C6 alkyl, R ₂ is C7 alkyl, R ₂ is C8 alkyl, R ₂ is C9 alkyl, R ₂ is C10 alkyl, R In _a further preferred embodiment, _R2 is _a C10 alkyl group, and in a further preferred embodiment, _{R2 is a} C11 alkyl group.

In a preferred embodiment, R ₂ is selected from C1-C14 straight chain alkyl, for example, R ₂ is selected from C2-C14 straight chain alkyl, R ₂ is selected from C3-C14 straight chain alkyl, R ₂ is selected from C4-C14 straight chain alkyl, R ₂ is selected from C5-C14 straight chain alkyl, R ₂ is selected from C6-C14 straight chain alkyl, R ₂ is selected from C7-C14 straight chain alkyl, R ₂ is selected from C8-C14 straight chain alkyl, R ₂ is selected from C9-C14 straight chain alkyl, R ₂ is selected from C10-C14 straight chain alkyl, C10-C13 straight chain alkyl, C10-C12 straight chain alkyl, C10-C11 straight chain alkyl, for example, R ₂ is C1 alkyl, R ₂ is C2 alkyl, R ₂ is C3 straight chain alkyl, R ₂ is C4 straight chain alkyl, R ₂ is straight chain C5 alkyl, R ₂ is C6 straight chain alkyl, R In a further preferred embodiment, _{R 2} is a C7 straight chain alkyl, R ₂ is a C8 straight chain alkyl, R ₂ is a C9 straight chain alkyl, R ₂ is a C10 straight chain alkyl, R ₂ is a C11 straight chain alkyl, R ₂ is a C12 straight chain alkyl, R ₂ is a C13 straight chain alkyl, and R ₂ is a C14 straight chain alkyl. In a further preferred embodiment, R ₂ is a C10 straight chain alkyl, and in a further preferred embodiment, R ₂ is a C11 straight chain alkyl.

In a preferred embodiment, in the compounds of the present invention, R ₃ is selected from C1-C14 alkyl, for example, R ₃ is selected from C2-C14 alkyl, R ₃ is selected from C3-C14 alkyl, R ₃ is selected from C4-C14 alkyl, R ₃ is selected from C5-C14 alkyl, R ₃ is selected from C6-C14 alkyl, R ₃ is selected from C7-C14 alkyl, R ₃ is selected from C8-C14 alkyl, R ₃ is selected from C8-C13 alkyl, R ₃ is selected from C8-C12 alkyl, R ₃ is selected from C8-C11 alkyl, R ₃ is selected from C8-C10 alkyl, R ₃ is selected from C8-C9 alkyl, for example, R ₃ is C1 alkyl, R ₃ is C2 alkyl, R ₃ is C3 alkyl, R ₃ is C4 alkyl, R ₃ is C5 alkyl, R ₃ is C6 alkyl, R ₃ is C7 alkyl, R ₃ is C8 alkyl, R ₃ is C9 alkyl, R _{R 3} is C10 alkyl, R ₃ is C11 alkyl, R ₃ is C12 alkyl, R ₃ is C13 alkyl, R ₃ is C14 alkyl. In a preferred embodiment, R ₃ is C8 alkyl.

In a preferred embodiment, R ₃ is selected from C1-C14 straight chain alkyl, for example, R ₃ is selected from C2-C14 straight chain alkyl, R ₃ is selected from C3-C14 straight chain alkyl, R ₃ is selected from C4-C14 straight chain alkyl, R ₃ is selected from C5-C14 straight chain alkyl, R ₃ is selected from C6-C14 straight chain alkyl, R ₃ is selected from C7-C14 straight chain alkyl, R ₃ is selected from C8-C14 straight chain alkyl, R ₃ is selected from C8-C13 straight chain alkyl, R ₃ is selected from C8-C12 straight chain alkyl, R ₃ is selected from C8-C11 straight chain alkyl, R ₃ is selected from C8-C10 straight chain alkyl, R ₃ is selected from C8-C9 straight chain alkyl, for example, R ₃ is C1 alkyl, R ₃ is C2 alkyl, R ₃ is C3 straight chain alkyl, R ₃ is C4 straight chain alkyl, R ₃ is straight chain C5 alkyl, R _{R 3} is a C6 straight chain alkyl, R ₃ is a C7 straight chain alkyl, R ₃ is a C8 straight chain alkyl, R ₃ is a C9 straight chain alkyl, R ₃ is a C10 straight chain alkyl, R ₃ is a C11 straight chain alkyl, R ₃ is a C12 straight chain alkyl, R ₃ is a C13 straight chain alkyl, and R ₃ is a C14 straight chain alkyl. In a preferred embodiment, R ₃ is a C8 straight chain alkyl.

The present application further provides a nanoparticle composition comprising a lipid component, wherein the lipid component comprises a compound provided herein or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

In some embodiments, the nanoparticle composition has an average particle size of 50 nm to 110 nm. For example, it can be 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, 81nm, 82nm, 83nm, 84nm, 85nm, 86nm, 87nm, 88nm, 89nm, 90nm, 91nm, 92nm, 9 3nm, 94nm, 95nm, 96nm, 97nm, 98nm, 99nm, 100nm, 101nm, 102nm, 103nm, 104n m, 105nm, 106nm, 107nm, 108nm, 109nm.

The nanoparticle composition can include, for example, lipid nanoparticles (LNP), 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 aqueous compartments. The lipid bilayers can be functionalized and/or cross-linked to each other. The lipid bilayer can include one or more ligands, proteins or channels.

The nanoparticle compositions described herein include a lipid component, the lipid component including at least one compound according to formula (I) or (II). For example, the lipid component of the nanoparticle composition may include one or more of the compounds of the present invention. The nanoparticle composition may also include a variety of other components. For example, in addition to the compound according to formula (I), the lipid component of the nanoparticle composition may also include one or more other lipids.

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

The lipid component of the nanoparticle composition can include one or more structural lipids. The structural lipid can be selected from, but not limited to, cholesterol, coprostanol, sitosterol, ergosterol, stigmasterol and mixtures thereof, but not limited thereto. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid includes cholesterol and corticosteroids (e.g., prednisolone, dexamethasone, prednisone and hydrocortisone), or a combination thereof.

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

Dilauroyl phosphatidylcholine (DLPC),

Dimyristoylphosphatidylcholine (DMPC),

Dioleoylphosphatidylcholine (DOPC),

Dipalmitoylphosphatidylcholine (DPPC),

Distearoylphosphatidylcholine (DSPC),

Dioleoylphosphatidylcholine (DUPC),

Palmitoyloleoylphosphatidylcholine (POPC),

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

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

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

1,2-Divinyl-sn-glycero-3-phosphocholine,

1,2-Diaryl acyl-sn-glycero-3-phosphocholine,

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

1,2-Dihydroxytin-sn-glycerol-3-phosphoethanolamine (ME 16.0PE),

1,2-Distearoyl-sn-glycero-3-phosphoethanolamine,

1,2-Divinyl alcohol-sn-glycero-3-phosphoethanolamine,

1,2-Divinyl-sn-glycero-3-phosphoethanolamine,

1,2-Diaryl-sn-glycero-3-phosphoethanolamine,

1,2-dithiohexaenoic acid-sn-glycero-3-phosphoethanolamine,

1,2-Diol-sn-glycero-3-phospho-(1-glycero) sodium salt (DOPG) or sphingomyelin.

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.

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

The nanoparticle compositions may comprise one or more therapeutic and/or prophylactic agents.

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

The therapeutic and/or prophylactic agents include biologically active substances and may alternatively be referred to as "active agents". A therapeutic and/or prophylactic agent may be a substance that, once delivered to a cell or organ, produces a desired change in a 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, a therapeutic and/or prophylactic agent is a small molecule drug that can be used to treat a specific disease, disorder, or condition. Examples of drugs that can be used in the nanoparticle compositions include, but are not limited to, antineoplastic agents (e.g., vincristine, doxorubicin, mitoxantrone, camptothecin, cisplatin, bleomycin, cyclophosphamide, methotrexate, and streptozotocin), antitumor 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-infective agents, local anesthetics (e.g., dibucaine and chlorpromazine), beta-adrenergic blockers (e.g., propranolol, timolol, and laprostatin), and betadalafil), antihypertensives (e.g., clonidine and hydralazine), antidepressants (e.g., imipramine, amitriptyline, and doxepin), anticonversion agents (e.g., phenytoin), antihistamines (e.g., diphenhydramine, chlorpheniramine, and promethazine), antibiotics/antibacterials (e.g., gentamicin, ciprofloxacin, and cefoxitin), antifungals (e.g., miconazole, terconazole, econazole, isoconazole, butoconazole, clotrimazole, itraconazole, nystatin, and naftifine), antiparasitics, hormones, hormone antagonists, immunomodulators, neurotransmitter antagonists, antiglaucoma agents, vitamins, anesthetics, and imaging agents.

In some embodiments, the therapeutic and/or preventive is a cytotoxin, a radioactive ion, a chemotherapeutic agent, a vaccine, a compound that causes an immune response, and/or another therapeutic and/or preventive. 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, doxorubicin, daunorubicin, dihydroxyanthraquinone, ketomethoate, 1-nortestosterone, oryzalin, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoid alkaloids, e.g., maytansinol, racletin (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, sa 153 and. Vaccines include compounds and formulations capable of providing immunity to one or more conditions associated with infectious diseases (e.g., influenza, measles, human papillomavirus (HPV), rabies, meningitis, pertussis, tetanus, tetanus, plague, hepatitis and tuberculosis), including mRNA encoding antigens and/or epitopes derived from infectious diseases. Vaccines also include compounds and formulations that direct an immune response against cancer cells, and may include mRNA encoding tumor cell-derived antigens, epitopes and/or neo-epitopes. Compounds that elicit an immune response may include vaccines, corticosteroids (e.g., dexamethasone), and other species.

Other therapeutic and/or prophylactic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarboxylase), alkylating agents (e.g., methylchlorothiazide, thiotepa-chloramphenicol, racelotomycin (CC-1065), melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunorubicin) and doxorubicin), antibiotics (e.g., dacomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and antimitotic agents (e.g., vincristine, vinblastine, paclitaxel, and zeaxanthin)

In some embodiments, vaccines and/or compounds capable of eliciting an immune response are administered intramuscularly via a composition comprising a compound of Formula (I), (II), such as one or more of Compounds 1-14. Other therapeutic and/or preventive measures include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarboxyzine), alkylating agents (e.g., methoxazole, thiopyradex, chlorambucil (CC-1065), melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin, melamicin, doxorubicin, actinomycin), anthracyclines (AMC)), and antimitotic agents (e.g., vincristine, vinblastine, paclitaxel, and maytansinoids.

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

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 bound or can be bound to an oligonucleotide chain. The exemplary polynucleotides used according to the present invention include, but are not limited to, deoxyribonucleic acid (DNA), ribonucleic acid (RNA) including messenger mRNA (mRNA), its hybrid, RNAi inducer, RN-Ai agent, siRNA, shRNA, miRNA, antisense RNA, ribozyme, catalytic DNA, RNA that induces triple helix formation, aptamer, carrier, etc. One or more. In some embodiments, therapeutic and/or preventive is RNA. The RNA that can be used for the compositions and methods described herein can be selected from, but is not limited to, short polymers, antigametocytes, antisense, ribozymes, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), double-stranded RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA) and mixtures thereof. In certain embodiments, RNA is mRNA.

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

In other embodiments, the therapeutic and/or preventive agent is siRNA. siRNA may be able to selectively knock down or down-regulate the expression of a target gene. For example, siRNA can be selected to silence genes associated with a specific disease, illness or condition when a nanoparticle composition comprising siRNA is administered to a subject in need thereof. siRNA may comprise a sequence complementary to an mRNA sequence encoding a gene of interest or protein. In some embodiments, siRNA may be an immunomodulatory siRNA.

In some embodiments, the therapeutic and/or preventive agent is a shRNA or a vector or plasmid encoding the shRNA. After the appropriate construct is delivered to the nucleus, the shRNA can be produced inside the target cell. The construct and mechanism associated with shRNA are well known in the relevant art.

In addition to those described in the preceding sections, the nanoparticle compositions of the present application may further comprise one or more components. For example, the nanoparticle compositions may comprise one or more small hydrophobic molecules, such as vitamins (e.g., vitamin A or vitamin E) or sterols.

In addition to the above components, the nanoparticle composition can include any substance useful in a pharmaceutical composition. For example, the nanoparticle composition can include one or more pharmaceutically acceptable excipients or auxiliary ingredients, such as but not limited to one or more solvents, dispersion media, diluents, dispersing aids, suspending agents, granulation aids, disintegrants, fillers, glidants, liquid carriers, adhesives, surfactants, isotonic agents, thickeners or emulsifiers, buffers, lubricants, oils, preservatives and other substances. Excipients such as waxes, butters, colorants, coatings, flavorings and flavoring agents may also be included. Pharmaceutically acceptable excipients are well known in the art.

Nanoparticle compositions described herein can include lipid components and one or more other components, such as therapeutic and/or preventive agents. Nanoparticle compositions can be designed for one or more specific applications or targets. The elements of nanoparticle compositions can be selected based on specific applications or targets and/or based on the efficacy, toxicity, cost, ease of use, availability or other characteristics of one or more elements. Similarly, specific formulations of nanoparticle compositions can be selected for specific applications or targets based on the efficacy and toxicity of a specific combination of elements, for example.

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

The nanoparticle composition can be designed for one or more specific applications or targets. For example, a nanoparticle composition can be designed to deliver therapeutic and/or preventive such as RNA to specific cells, tissues, organs, or systems or groups thereof in a mammal. The physicochemical properties of the nanoparticle composition can be changed to increase selectivity for specific body targets. For example, the particle size can be adjusted based on the window size of different organs. The therapeutic and/or preventive agent included in the nanoparticle composition can also be selected based on one or more desired delivery targets. For example, a therapeutic and/or preventive agent can be selected for specific indications, conditions, diseases or disorders and/or for delivery (e.g., local or specific delivery) to specific cells, tissues, organs, or systems or groups thereof. In certain embodiments, a nanoparticle composition can include mRNA encoding a polypeptide of interest that can be translated intracellularly to produce a polypeptide of interest. Such a composition can be designed to be specifically delivered to a specific organ. In some embodiments, the composition can be designed to be specifically delivered to the liver of a mammal.

The amount of the therapeutic and/or prophylactic agent in the nanoparticle composition can depend on the size, composition, desired target and/or application or other properties of the nanoparticle composition and the properties of the therapeutic and/or/or prophylactic agent. For example, the amount of RNA that can be used in the nanoparticle composition can depend on the size, sequence and other characteristics of the RNA. The relative amounts of the therapeutic and/or prophylactic agent and other elements (e.g., lipids) in the nanoparticle composition can also vary.

The properties of the nanoparticle composition can depend on its components. For example, a nanoparticle composition comprising cholesterol as a structural lipid can have different properties than a nanoparticle composition comprising a different structural lipid. Similarly, the properties of a nanoparticle composition can depend on the absolute or relative amounts of its components. For example, a nanoparticle composition comprising a higher mole fraction of phospholipids can have different properties than a nanoparticle composition comprising a lower mole fraction of phospholipids. The properties can also vary depending on the method and conditions of preparation of the nanoparticle composition.

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

The average particle size of the nanoparticle composition is 50 nm-110 nm.

The nanoparticle composition can be relatively uniform. The polydispersity index can be used to indicate the uniformity of the nanoparticle composition, for example, the particle size distribution of the nanoparticle composition. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. The polydispersity index of the nanoparticle composition is 0.04-0.20. For example, it 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, 0.19.

The encapsulation efficiency of therapeutic and/or prophylactic agents describes the amount of therapeutic and/or prophylactic agents that are encapsulated or otherwise associated with the nanoparticle composition after preparation relative to the initial amount provided. The encapsulation efficiency is expected to be as high as possible (e.g., close to 100%). The encapsulation efficiency can be measured, for example, by comparing the amount of therapeutic and/or prophylactic agents in a solution containing a nanoparticle composition before and after decomposing 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 (e.g., RNA) in a solution. For the nanoparticle compositions described herein, the encapsulation efficiency of the therapeutic and/or prophylactic agent can be at least 50%, such as 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 can be at least 80%. In certain embodiments, the encapsulation efficiency can be at least 90%.

The nanoparticle composition may optionally include one or more coatings. For example, the nanoparticle composition may be formulated into a capsule, film, or tablet having a coating. The capsule, film, or tablet containing the composition described herein may have any useful size, tensile strength, hardness, or density.

The nanoparticle composition of the present application can be formulated as a pharmaceutical composition in whole or in part. The pharmaceutical composition may include one or more nanoparticle ingredients. For example, the pharmaceutical composition may include one or more nanoparticle compositions, which include one or more different therapeutic and/or preventive agents. The pharmaceutical composition may further include one or more pharmaceutically acceptable excipients or auxiliary ingredients, such as those described herein, unless any conventional excipients or auxiliary ingredients may be incompatible with one or more components of the nanoparticle composition. If an excipient or auxiliary ingredient is incompatible with a component of the nanoparticle composition, its combination with the component may cause any undesirable biological effect or other harmful effect.

In some embodiments, one or more excipients or auxiliary ingredients may account for more than 50% of the total mass or volume of the pharmaceutical composition including the nanoparticle composition. For example, one or more excipients or auxiliary ingredients may constitute 50%, 60%, 70%, 80%, 90% or more of pharmaceutical practice. In some embodiments, the 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.

According to the one or more nanoparticle compositions in the pharmaceutical composition of the present application, the relative amount of one or more pharmaceutically acceptable excipients and/or any other ingredients will vary, depending on the identity, size and/or disease condition of the subject being treated, and further depending on the route of administration of the composition. For example, the pharmaceutical composition may include 0.1% to 100% (wt/wt) of one or more nanoparticle compositions.

The nanoparticle compositions and/or pharmaceutical compositions comprising one or more of the nanoparticle compositions can be administered to any patient or subject, including those patients or subjects who may benefit from the therapeutic effect provided by delivering a therapeutic and/or preventive agent to one or more specific cells, tissues, organs, or systems or groups thereof (e.g., the renal system). Although the description of the nanoparticle compositions and pharmaceutical compositions comprising nanoparticle compositions provided herein is primarily directed to compositions suitable for administration to humans, it will be appreciated by those skilled in the art that such compositions are generally suitable for administration to any other mammal. It is well known that compositions suitable for human administration are modified to make the compositions suitable for administration to various animals, and such modifications can be designed and/or performed by a generally skilled veterinary pharmacologist with only ordinary (if any) experiments, including but not limited to humans, other primates, and other artificial animals, including commercially relevant mammals such as cows, pigs, hoses, sheep, cats, dogs, mice, and/or rats.

Pharmaceutical compositions comprising one or more nanoparticle compositions can be prepared by any method known or hereafter developed in the art of pharmacology. Generally, such preparation methods involve combining the active ingredient with an excipient and/or one or more other auxiliary ingredients and then, if desired or necessary, dividing, shaping and/or packaging the product into desired single or multiple dosage units.

The pharmaceutical composition according to the present application can be prepared, packaged and/or sold in batches of single unit doses and/or multiple single unit doses. As used herein, a "unit dose" is a precise amount of a pharmaceutical composition containing a predetermined amount of active ingredient (e.g., nanoparticle composition). The amount of the active ingredient is generally equal to the dosage of the active ingredient to be given to the subject and/or a convenient portion of the dosage, such as half or one-third of the dosage.

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 medicaments), injectable forms, solid dosage forms (e.g., capsules, tablets, tablets, etc.) (liquids, powders and granules), dosage forms for topical and/or transdermal administration (e.g., ointments, 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 medicaments.

In addition to the active ingredient, the liquid dosage form can include an inert diluent commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (particularly cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters and mixtures thereof. In addition to the inert diluent, the oral composition can also include other therapeutic agents and/or preventive agents, other agents, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and/or spices. In certain embodiments for parenteral administration, the composition is mixed with a solubilizer (e.g., alcohol, oil, modified oil, glycol, polysorbate, cyclodextrin, polymer and/or a combination thereof).

Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants, wetting agents and/or suspending agents according to known techniques. Sterile injectable preparations can be sterile injectable solutions, suspensions and/or emulsions in nontoxic parenteral acceptable diluents and/or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water. Sterile fixed oils are generally used as solvents or suspension media. For this reason, 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 injections.

The injectable formulations can be sterilized by, for example, filtration through a bacteria-retaining filter and/or by incorporating sterilizing agents in the form of sterile solid compositions prior to dissolution or dispersion in sterile water or other sterile injectable medium for use.

In order to prolong the effect of active ingredients, it is usually desirable to slow down the absorption of active ingredients from subcutaneous or intramuscular injections. This can be achieved by using a liquid suspension of a crystalline or amorphous substance with poor water solubility. Then, the absorption rate of the drug depends on its dissolution rate, which in turn depends on the crystal size and crystalline form. Alternatively, delayed absorption of the drug form administered parenterally is achieved by dissolving or suspending the drug in an oil carrier. The injectable drug depot form is made by forming a microcapsule matrix of the drug in a biodegradable polymer (such as polylactic acid-polyethylene glycol). Depending on the ratio of the drug to the polymer and the properties of the specific polymer used, the drug release rate can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Reserve injection preparations are prepared by embedding the drug in liposomes or microemulsions compatible with body tissues.

Compositions for rectal or vaginal administration are generally 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 ambient temperature but liquid at body temperature and therefore melts in the rectum or vaginal cavity to release the active ingredient.

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. starch, lactose, sucrose, glucose, mannitol and silicic acid), binders (e.g. carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic), humectants (e.g. glycerol), disintegrants (e.g. agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate), slow dissolving agents (e.g. paraffin), absorption promoters (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 glycol, sodium lauryl sulfate) and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may comprise buffering agents.

Solid compositions of similar type 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 glycols. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. They may optionally contain opacifiers and may have a composition that releases the active ingredient only, or preferably, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedded compositions that can be used include polymeric substances and waxes. Solid compositions of similar type can be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose and milk sugar and high molecular weight polyethylene glycols.

The dosage form for topical and/or transdermal administration of compositions may include ointment, paste, cream, lotion, gel, powder, solution, spray, inhalant and/or patch.Usually, active ingredient is aseptically mixed with pharmaceutically acceptable excipient and/or any preservative and/or buffer that may be needed.In addition, the application contemplates the use of transdermal patch, which generally has the additional advantage of providing controlled delivery of compound to health.Such dosage form can be prepared, for example, by dissolving the compound and/or being distributed in a suitable medium.Alternatively or additionally, rate can be provided by any of the following control rate control membrane and/or by dispersing the compound in a polymer matrix and/or gel.

Formulations suitable for topical administration include, but are not limited to, liquid and/or semi-liquid preparations, 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, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. The formulation for topical administration may further comprise one or more of the other ingredients described herein.

Pharmaceutical compositions can be prepared, packaged and/or sold in formulations suitable for pulmonary administration via the oral cavity. Such formulations may comprise dry particles of the active ingredient. Such compositions are conveniently in dry powder form for administration using a device comprising a dry powder reservoir and/or using a self-propelling solvent/powder dispensing container (e.g., a device comprising an active ingredient dissolved and/or suspended in a low-boiling propellant in a sealed container). Powder compositions may include a solid fine powder diluent, such as sugar, and are conveniently provided in unit dosage form.

Low boiling point propellants generally include liquid propellants having a boiling point below 65°F at atmospheric pressure. Typically, the propellant may comprise 50% to 99.9% (wt/wt) of the composition, while the active ingredient may comprise 0.1% to 20% (wt/wt) of the composition. The propellant may further comprise other ingredients, such as liquid nonionic and/or solid anionic surfactants and/or solid diluents (whose particle size may be of the same order of magnitude as the particles comprising the active ingredient).

The pharmaceutical composition prepared for pulmonary delivery can provide the active ingredient in the form of droplets of solution and/or suspension. Such preparations can be prepared, packaged and/or sold as aqueous and/or diluted alcohol solutions and/or suspensions, optionally sterile, containing active ingredients, and can be conveniently administered using any atomization and/or atomization device. Such preparations may further include one or more other ingredients, including but not limited to flavoring agents such as saccharin sodium, 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 1nm to 200nm.

The formulations described herein for pulmonary delivery can be used for intranasal delivery of pharmaceutical compositions. Another formulation suitable for intranasal administration is a coarse powder comprising the active ingredient and having an average particle size of 0.2 μm to 500 μm. Such a formulation is administered in the manner of taking snuff, i.e., by rapid inhalation through the nasal passages from a powder container held near the nose.

Preparations suitable for nasal administration may, for example, contain 0.1% (wt/wt) to 100% (wt/wt) of active ingredients, and may contain one or more other active ingredients described herein. Pharmaceutical compositions may be prepared, packaged and/or sold in preparations suitable for buccal administration. Such preparations may, for example, be in the form of tablets and/or lozenges prepared using conventional methods, and may, for example, be 0.1% to 20% (wt/wt) of active ingredients, the remainder comprising an oral soluble and/or degradable composition, and optionally, one or more other ingredients described herein. Alternatively, preparations suitable for oral administration may include powders and/or atomized and/or spray solutions and/or suspensions containing active ingredients. Such powdered, aerosolized and/or aerosolized preparations may have an average particle and/or droplet size in the range of 0.1nm to 200nm when dispersed, and may further include one or more other ingredients described herein.

The pharmaceutical composition can be prepared, packaged and/or sold in the form of a formulation suitable for ophthalmic administration. Such a formulation can be, for example, in the form of drops, including, for example, a 0.1/1.0% (wt/wt) solution and/or suspension of the active ingredient in an aqueous or oily liquid excipient. Such drops may further contain a buffer, a salt and/or one or more of any other additive ingredients described herein. Other formulations that can be used for ophthalmic administration include formulations containing the active ingredient in microcrystalline form and/or a liposomal formulation. Ear drops and/or eye drops are considered within the scope of the present disclosure Methods for producing polypeptides in cells

The application further provides a method for producing a target polypeptide in a mammalian cell. The method for producing the polypeptide comprises contacting the cell with a nanoparticle composition comprising an mRNA encoding the target polypeptide. After contacting the cell with the nanoparticle composition, the mRNA can be absorbed and translated in the cell to produce the target polypeptide.

Typically, the step of contacting mammalian cells with a nanoparticle composition comprising an mRNA encoding a polypeptide of interest can be performed in vivo, in vitro, in culture or in vitro. The amount of the nanoparticle composition contacted with the cell and/or the amount of mRNA therein can depend on the type of cells or tissues contacted, the mode of administration, the physicochemical properties of the nanoparticle composition and mRNA (e.g., size, charge and chemical composition) and other factors. Typically, an effective amount of the nanoparticle composition will allow effective production of polypeptides in cells. Metrics of efficiency may include polypeptide translation (represented by polypeptide expression), mRNA degradation levels and immune response indicators.

The step of contacting the nanoparticle composition including the mRNA with the cell may involve or cause transfection. The phospholipids included in the lipid component of the nanoparticle composition can promote transfection and/or increase transfection efficiency, for example, by interacting and/or fusing with the cell or intracellular membrane. Transfection can allow translation of the intracellular mRNA.

In some embodiments, the nanoparticle compositions described herein can be used for treatment. For example, the mRNA contained in the nanoparticle composition can encode a therapeutic polypeptide (e.g., in a translatable region) and produce a therapeutic polypeptide when contacting and/or entering (e.g., transfecting) a cell. In other embodiments, the mRNA contained in the nanoparticle composition can encode a polypeptide that can improve or increase the immunity of a subject. For example, the mRNA may encode granulocyte colony stimulating factor or trastuzumab.

In certain embodiments, the mRNA included in the nanoparticle composition can encode a recombinant polypeptide that can replace one or more polypeptides that may be substantially absent in the cell in contact with the nanoparticle composition. Due to genetic mutations of the encoding gene or its regulatory pathway, one or more substantially absent polypeptides may be lacking. Alternatively, the recombinant polypeptide produced by mRNA translation can antagonize the activity of endogenous proteins present in the cell, on the cell surface, or secreted from the cell. Antagonistic recombinant polypeptides may be ideal to counteract the harmful effects caused by the activity of endogenous proteins, such as localization caused by activity changes or mutations. In another alternative, the recombinant polypeptide produced by mRNA translation can indirectly or directly antagonize the activity of biological parts present in the cell, on the cell surface, or secreted from the cell. Antagonistic biological parts may include, but are not limited to, lipids (e.g., cholesterol), lipoproteins (e.g., low-density lipoproteins), nucleic acids, carbohydrates, and small molecule toxins. The recombinant polypeptide produced by mRNA translation can be engineered to be positioned in the cell, such as in a specific compartment such as the nucleus, or can be engineered to be secreted from the cell or transported to the plasma membrane of the cell.

In some embodiments, contacting a cell with a nanoparticle composition comprising mRNA can reduce the innate immune response of the cell to exogenous nucleic acids. The cell can be contacted with a first nanoparticle composition comprising a first amount of a first exogenous mRNA, the first exogenous mRNA comprising a translatable region, and the level of the innate immune response of the cell to the first exogenous mRNA can be determined. Subsequently, the cell can be contacted with a second composition comprising a second amount of a first exogenous mRNA, the second amount being a first exogenous mRNA of a smaller amount than the first amount. Alternatively, the second composition can comprise a second exogenous mRNA of a first amount that is different from the first exogenous mRNA. The step of contacting the cell with the first and second compositions can be repeated one or more times. In addition, the efficiency of polypeptide production (e.g., translation) in the cell can be optionally determined, and the cell can be repeatedly contacted with the first and/or second compositions again until the target protein production efficiency is reached.

The present application further provides a method for treating a disease or condition in a mammal, the method comprising administering to a mammal a therapeutically effective amount of any of the foregoing nanoparticle compositions, in particular, the nanoparticle composition can be used to treat a disease, condition or symptom characterized by a lack or abnormality in protein or polypeptide activity. For example, a nanoparticle composition comprising an mRNA encoding a missing or abnormal polypeptide can be administered or delivered to a cell. Subsequent translation of the mRNA can produce a polypeptide, thereby reducing or eliminating problems caused by a lack or abnormal activity caused by the polypeptide. Because translation may occur rapidly, these methods and compositions can be used to treat acute diseases, conditions or conditions such as sepsis, stroke and myocardial infarction. The therapeutic and/or preventive agents included in the nanoparticle composition may also be able to change the transcription rate of a given species, thereby affecting gene expression.

Diseases, disorders and/or conditions characterized by malfunctioning or aberrant protein or polypeptide activity include, but are not limited to, rare diseases, infectious diseases (such as vaccines and therapeutics), cancer and proliferative diseases, genetic diseases (e.g., cystic fibrosis), autoimmune diseases, diabetes, neurodegenerative diseases, cardiovascular and renal vascular diseases, and metabolic diseases.

The present application provides methods involving the administration of nanoparticle compositions comprising one or more therapeutic and/or preventive agents and pharmaceutical compositions comprising nanoparticle compositions. With respect to the features and embodiments of the present application, the terms therapeutic and preventive can be used interchangeably herein. The therapeutic composition or its imaging, diagnostic or preventive composition can be administered to a subject using any reasonable amount and any route of administration that is effective for preventing, treating, diagnosing or imaging a disease, disorder and/or condition and/or any other purpose. The specific dosage for a particular subject may vary depending on the type, age and general condition of the subject; the purpose of management; the specific ingredients; the mode of administration; and the like. The compositions according to the present application can be formulated in unit dosage form for ease of administration and uniform dosage. However, it should be understood that the total daily dosage of the composition of the present application will be determined by the attending physician within the scope of reasonable medical judgment. The specific therapeutically effective, prophylactically effective, or other appropriate dosage level (e.g., for imaging) for any particular patient will depend on a variety of factors, including the severity and identification of the disease being treated, if any; the therapeutic and/or prophylactic agent(s) employed; the specific ingredient employed; the age, weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific pharmaceutical ingredient employed; the duration of the treatment; drugs used in combination or concomitantly with the specific pharmaceutical ingredient employed; and factors well known in the medical community.

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

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

As another example of targeted or specific delivery, mRNA encoding a protein binding partner (e.g., an antibody or its functional fragment, scaffold protein or peptide) or a receptor on the cell surface can be included in the nanoparticle composition. The mRNA may be used to increase the mountain or alternatively to direct the synthesis and extracellular localization of lipids, carbohydrates or other biological parts. Alternatively, other therapeutic and/or preventive agents or elements (e.g., lipids or ligands) of the nanoparticle composition can be selected based on their affinity for specific receptors (e.g., low-density lipoprotein receptors) so that the nanoparticle composition can more easily interact with the target cell population including the receptor.

In some embodiments, the ligand can be a surface-bound antibody, which can allow for the modulation of cell targeting specificity. This is particularly useful because highly specific antibodies can be generated against a target epitope of a desired targeting site. In one embodiment, multiple antibodies are expressed on the cell surface, and each antibody can have a different specificity for the desired target. This approach can increase the affinity and specificity of the targeting interaction.

Targeted cells can 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, cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticular cells, leukocytes, granulocytes, and tumor cells.

In some embodiments, nanoparticle compositions may be targeted to hepatocytes.

The nanoparticle compositions of the present application can be administered by any route. In some embodiments, a composition comprising a preventive, diagnostic or imaging composition is administered by one or more of a variety of routes, including oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal or intradermal, intradermal, rectal, intravaginal, intraperitoneal, intraocular, subretinal, intravitreal, topical (e.g., by powder, ointment, cream, gel, lotion and/or drops), mucosal, nasal, buccal, intestinal, vitreous, intratumoral, sublingual, intranasal; by intratracheal instillation, bronchial instillation and/or inhalation; as an oral spray and/or powder, nasal spray and/or aerosol, and/or through a portal vein catheter. In some embodiments, the composition can be administered intravenously, intramuscularly, intradermally, intraarterially, intratumorally, subcutaneously, intraocularly, subretinal, intravitreal or by inhalation. However, in view of possible progress in drug delivery science, the present application encompasses delivery or description of compositions described herein by any suitable route. Generally, the most suitable route of administration depends on a variety of factors, including the properties of the nanoparticle composition, including one or more treatments and/or preventive measures (e.g., its stability in various body environments (e.g., blood and gastrointestinal tract)), the patient's condition (e.g., whether the patient can tolerate a specific route of administration), etc.

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

Nanoparticle compositions comprising one or more therapeutic and/or preventive agents can be used in combination with one or more other therapeutic, preventive, diagnostic or imaging agents. "In combination with..." does not mean that the agents must be administered and/or formulated to be delivered together at the same time, although these delivery methods are within the scope of the present application. For example, one or more nanoparticle compositions comprising one or more different therapeutic and/or preventive agents can be administered in combination. The composition can be administered simultaneously, before or after one or more other desired treatments or medical procedures. Typically, each agent will be administered at a dose and/or schedule determined for the agent. In some embodiments, the present application includes delivering a composition or its imaging, diagnostic or preventive composition in combination with an agent that improves its bioavailability, reduces and/or changes its metabolism, inhibits its excretion and/or regulates contraction in the body.

It should be understood that the therapeutic, preventive, diagnostic or imaging agents used in combination can be administered together in a single composition or separately in different compositions. Generally, it is desirable that the level of use of the agents used in combination does not exceed the level at which they are used alone. In some embodiments, the level of combined use may be lower than the level used alone.

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

In some embodiments, the method for treating a subject in need or delivering a therapeutic and/or preventive drug to a subject (e.g., a mammal) may include pre-treating the subject with one or more medicaments before administering the nanoparticle composition. For example, the subject may be pre-treated with an effective amount of (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, H1 receptor blocker or H2 receptor blocker. Pre-treatment may occur 24 hours or less (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) before administering the nanoparticle composition, and may occur once, twice or more times, for example, in the case of increasing the dosage.

Example

The present invention is described below in conjunction with specific examples. These examples are not intended to limit the scope of the present invention, but to provide guidance for those skilled in the art to prepare and use the compounds and compositions of the present invention.

The synthetic routes of the compounds of this embodiment are as follows:

Example 1

The synthetic route is as follows:

To a DCM (6 mL) solution of compound A-1 (570 mg, 2.92 mmol), add B (1350 mg, 3.49 mmol) and DCC (910 mg, 4.41 mmol), stir at 25 °C for 15 hours, monitor the reaction by TLC, filter after completion of the reaction, concentrate, and perform silica gel column chromatography (PE:EA=50:1) to obtain compound C-1 (1510 mg, yield 92%).

To the ethanol solution (3 mL) of compound E-1 (1000 mg, 2.17 mmol), add 10 times the equivalent of compound D-1 (2540 mg, 21.67 mmol) and DIEA (840 mg, 6.50 mmol), heat at 75 ° C for 24 hours, monitor the reaction by TLC, and cool to room temperature after the reaction is complete. Add water to the reaction solution for dilution, extract with ethyl acetate (50 mL × 3), concentrate, and perform silica gel column chromatography (PE: EA = 5: 1) to obtain oily compound F-1 (659 mg, yield 61%)

To a mixed solvent of acetonitrile and cyclopentyl methyl ether (cyclopentyl methyl ether: acetonitrile = 1:1, 4 mL) of compound C-1 (300 mg, 0.53 mmol), 1.0 equivalent of compound F-1 (270 mg, 0.53 mmol), K2CO3 (220 mg, 1.59 mmol) and KI (200 mg, 1.20 mmol) were added, and the mixture was heated at 75°C for 24 hours. The reaction was monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature. Water was added to the reaction solution for dilution, and the mixture was extracted with ethyl acetate (30 mL × 3), concentrated, and subjected to silica gel column chromatography (dichloromethane: methanol = 20:1) to ^obtain an oily compound 1 (234 mg, yield 45%). NMR(400MHz, CDCl3)δ5.42(d,J＝5.0Hz,1H),4.91(p,J＝6.3Hz,1H),4.65(ddt,J＝11.5,8.0,4.3Hz,1H),3.69(q,J＝6.0Hz,2H),2.56(s,6H),2.37–2.30(m,6H), 2.09–1.99(m,2H),1. 89(ddt,J＝13.9,9.3,4.4Hz,4H),1.76–1.49(m,11H),1.33(d,J＝24.4Hz,54H),1.17(qd,J＝10.8,4.4Hz,6H),1.06(s,4H),0.98–0.85(m,15H),0.72(s ,3H).MS-ESI(m/z):981(M+H) ⁺ .

Example 2

The preparation method is the same as compound 1. Using 6-bromohexanoic acid, ethanolamine, cholesterol, heptadecan-9-yl 8-bromooctanoate as raw materials, oily compound 2 can be prepared. ¹ H NMR (400MHz, CDCl3) δ5.41 (d, J = 4.9 Hz, 1H), 4.90 (p, J = 6.2 Hz, 1H), 4.65 (ddt, J = 11.5, 8.2, 4.3 Hz, 1H), 3.69 (s, 2H), 2.70 (d, J = 48.8 Hz, 6H), 2.40-2.25 (m, 6H), 2.13-1.96 (m, 2 H),1.89(ddt,J＝14.2,9.7,4.7Hz,4H),1.74–1.46(m,9H),1.33(d,J＝24.3Hz,48H),1.22–1.10(m,6H),1.06(s,4H),0.98–0.86(m,15H),0.72(s,3H). MS-ESI(m/z):925.2(M+H) ⁺ .

Example 3

The preparation method is the same as compound 1. Using 6-bromohexanoic acid, ethanolamine, cholesterol, and 6-bromohexanoic acid undecyl ester as raw materials, an oily compound 3 can be prepared. ¹ H NMR (400 MHz, CDCl3) δ5.46–5.38 (m, 1H), 4.65 (dtd, J＝11.6, 8.6, 4.3 Hz, 1H), 4.10 (t, J＝6.8 Hz, 2H), 3.70 (s, 2H), 2.72 (d, J＝43.8 Hz, 6H), 2.40–2.27 (m, 6H), 2.10–1.97 (m, 2H), 1 .89(ddt,J＝14.4,9.7,4.6Hz,4H),1.75–1.56(m,11H),1.46–1.24(m,32H),1.16(dd,J＝9.9,4.4Hz,6H),1.06(s,4H),0.97–0.87(m,12H),0.72(s,3H) .MS-ESI(m/z):813.0(M+H) ⁺ .

Example 4

The preparation method is the same as compound 1. Using 6-bromohexanoic acid, 6-amino-1-hexanol, cholesterol, and 6-bromohexanoic acid undecyl ester as raw materials, an oily compound 4 can be obtained. ¹ H NMR (400 MHz, CDCl3) δ5.41 (d, J = 5.1 Hz, 1H), 4.65 (dtd, J = 11.6, 8.6, 4.3 Hz, 1H), 4.09 (t, J = 6.8 Hz, 2H), 3.68 (t, J = 6.4 Hz, 2H), 2.61 (s, 6H), 2.39–2.27 (m, 6H), 2.09–1.96 (m, 2H), 1.8 8(ddt,J＝14.1,9.5,4.6Hz,4H),1.74–1.48(m,13H),1.33(d,J＝26.3Hz,38H),1.16(tt,J＝14.1,7.2Hz,6H),1.06(s,4H),0.97–0.86(m,12H),0.71(s, 3H).MS-ESI(m/z):869.0(M+H) ⁺ .

Example 5

The preparation method is the same as compound 1. Using 8-bromooctanoic acid, 6-amino-1-hexanol, cholesterol, and 6-bromohexanoic acid undecyl ester as raw materials, an oily compound 5 can be obtained. ¹ H NMR (400 MHz, CDCl3) δ5.42 (d, J = 5.0 Hz, 1H), 4.73–4.57 (m, 1H), 4.10 (t, J = 6.8 Hz, 2H), 3.70 (t, J = 6.2 Hz, 2H), 3.02 (d, J = 9.4 Hz, 6H), 2.44–2.26 (m, 6H), 2.04 (t, J = 15.3 Hz, 2H) ,1.94–1.77(m,4H),1.76–1.58(m,13H),1.57–1.26(m,42H),1.17(tt,J＝13.4,7.0Hz,6H),1.07(d,J＝2.3Hz,4H),0.99–0.87(m,12H),0.72(s,3H). MS-ESI(m/z):897.1(M+H) ⁺ .

Example 6

The preparation method is the same as compound 1. Using 8-bromooctanoic acid, 6-amino-1-hexanol, cholesterol, and heptadecan-9-yl 8-bromooctanoate as raw materials, an oily compound 6 can be prepared. ¹ H NMR (400 MHz, CDCl3) δ5.41 (d, J = 5.0 Hz, 1H), 4.90 (t, J = 6.2 Hz, 1H), 4.71–4.59 (m, 1H), 3.68 (t, J = 6.6 Hz, 2H), 2.52 (s, 6H), 2.39–2.27 (m, 6H), 2.10–1.97 (m, 2H), 1.88 (ddd, J ＝13.8,9.3,4.9Hz,4H),1.69–1.47(m,11H),1.32(d,J＝23.3Hz,58H),1.16(dd,J＝9.9,4.4Hz,6H),1.06(s,4H),0.96–0.86(m,15H),0.72(s,3H).MS-ESI (m/z):1009.2(M+H) ⁺ .

Example 7

The preparation method is the same as compound 1. Using 8-bromooctanoic acid, ethanolamine, cholesterol, and heptadecan-9-yl 8-bromooctanoate as raw materials, oily compound 7 can be prepared. ¹ H NMR (400 MHz, CDCl3) δ5.45–5.38 (m, 1H), 4.90 (p, J = 6.3 Hz, 1H), 4.65 (ddt, J = 11.4, 8.1, 4.3 Hz, 1H), 3.67 (t, J = 5.1 Hz, 2H), 2.75 (d, J = 6.5 Hz, 2H), 2.63 (d, J = 8.0 Hz, 4H), 2.40–2.26 (m, 6H ),2.10–1.96(m,2H),1.90(d,J＝13.6Hz,4H),1.72–1.48(m,9H),1.33(d,J＝24.7Hz,52H),1.22–1.10(m,6H),1.06(s,4H),0.97–0.88(m,15H),0.72( s,3H).MS-ESI(m/z):953.2(M+H)+.

Example 8

The preparation method is the same as compound 1. Using 8-bromooctanoic acid, ethanolamine, cholesterol, and 6-bromohexanoic acid undecyl ester as raw materials, oily compound 8 can be prepared. ¹ H NMR (400MHz, CDCl3) δ5.44–5.38 (m, 1H), 4.65 (ddt, J = 11.4, 8.0, 4.2 Hz, 1H), 4.09 (t, J = 6.8 Hz, 2H), 3.64 (t, J = 5.2 Hz, 2H), 2.71 (t, J = 5.2 Hz, 2H), 2.58 (p, J = 4.9 Hz, 4H), 2.39–2.27 (m, 6H), 2.09–1. 96(m,2H),1.88(ddd,J＝14.0,9.4,4.9Hz,4H),1.73–1.46(m,11H),1.43–1.25(m,36H),1.17(ddd,J＝19.5,9.7,5.9Hz,6H),1.06(s,4H),0.95–0.88(m, 12H),0.71(s,3H).MS-ESI(m/z):841.1(M+H) ⁺ .

Embodiment 9

The preparation method is the same as compound 1. Using 4-bromobutyric acid, 6-amino-1-hexanol, cholesterol, and heptadecan-9-yl 8-bromooctanoate as raw materials, oily compound 9 can be prepared. ¹ H NMR (400MHz, CDCl3) δ5.41 (d, J＝4.9 Hz, 1H), 4.90 (t, J＝6.2 Hz, 1H), 4.65 (dtd, J＝11.6, 8.5, 4.2 Hz, 1H), 3.69 (t, J＝6.5 Hz, 2H), 2.50 (s, 6H), 2.34 (dt, J＝17.5, 6.9 Hz, 6H), 2.12–1.9 6(m,2H),1.87(tdd,J＝13.1,9.4,4.9Hz,4H),1.71–1.47(m,11H),1.46–1.24(m,50H),1.22–1.11(m,6H),1.06(s,4H),0.98–0.87(m,15H),0.72(s, 3H).MS-ESI(m/z):953.3(M+H) ⁺ .

Example 10

The preparation method is the same as compound 1. Using 4-bromobutyric acid, ethanolamine, cholesterol, and heptadecan-9-yl 8-bromooctanoate as raw materials, an oily compound 10 can be prepared. ¹ H NMR (400 MHz, CDCl3) δ5.41 (dd, J=4.9,1.8 Hz,1H), 4.90 (p, J=6.3 Hz,1H), 4.66 (dtd, J=11.5,8.4,4.2 Hz,1H), 3.60 (t, J=5.2 Hz,2H), 2.65 (t, J=5.2 Hz,2H), 2.53 (dt, J=19.5,6.7 Hz,4H), 2.33 (dt, J=1 1.4,7.6Hz,6H),2.10–1.96(m,2H),1.95–1.76(m,4H),1.71–1.44(m,9H),1.43–1.23(m,44H),1.16(dd,J＝10.0,4.3Hz,6H),1.06(s,4H),0.97–0.85 (m,15H),0.72(s,3H).MS-ESI(m/z):897.1(M+H) ⁺ .

Embodiment 11

To a solution of compound C-3 (500 mg, 0.93 mmol) in acetonitrile (4 mL), add 0.45 equivalents of 6-amino-1-hexanol (50 mg, 0.42 mmol), K2CO3 (180 mg, 1.30 mmol) and KI (160 mg, 0.96 mmol), heat at 75 ° C for 24 hours, monitor the reaction by TLC, and cool to room temperature after the reaction is complete. Add water to the reaction solution for dilution, extract with ethyl acetate (30 mL × 3), concentrate, and silica gel column chromatography (dichloromethane: methanol = 30: 1) to obtain oily compound 11 (215 mg, yield 50%) ¹ H NMR (400MHz, CDCl3) δ5.45–5.38(m,2H),4.65(dt,J＝8.1,3.2Hz,2H),3.68(t,J＝6.5Hz,2H),2.47(s,4H),2.34(dd,J＝7.4,3.5Hz,8H),2.09–1.96(m,4H),1.94 –1.74(m,10H),1.70– 1.46(m,16H),1.45–1.24(m,22H),1.16(ddd,J＝18.9,11.0,4.2Hz,12H),1.06(s,8H),0.95(d,J＝6.5Hz,6H),0.90(dd,J＝6.6,1.9Hz,12H),0.72(s,6H) .MS-ESI(m/z):1027.2(M+H) ⁺ .

Example 12

The preparation method is the same as compound 11. White solid compound 12 can be prepared using 4-bromobutyric acid, ethanolamine and cholesterol as raw materials. ¹ H NMR (400MHz, CDCl3) δ5.48–5.39 (m, 2H), 4.67 (dt, J＝7.9, 3.5Hz, 2H), 3.65 (s, 2H), 2.66 (d, J＝39.2Hz, 4H), 2.37 (dd, J＝7.8, 5.2Hz, 8H), 2.15–1.98 (m, 4H), 1.91 (ddd, J＝13.4, 9.0, 4.8Hz, 10H ),1.70–1.48(m,14H),1.40(d,J＝8.2Hz,16H),1.18(ddd,J＝19.0,11.1,4.3Hz,12H),1.08(s,8H),0.98(d,J＝6.4Hz,6H),0.93(dd,J＝6.6,1.8Hz,12H),0.7 4(s,6H).MS-ESI(m/z):971.2(M+H) ⁺ .

Embodiment 13

The preparation method is similar to the above compound.

Embodiment 14

The preparation method is similar to the above compound.

Embodiment 15

The preparation method is the same as compound 1, using 4-bromobutyric acid, ethanolamine, cholesterol, 6-bromohexanoic acid undecyl ester as raw materials to obtain oily compound 15, ¹ H NMR (400MHz, CDCl ₃ )δ4.08(t,J＝6.8Hz,2H),3.58(t,J＝5.3Hz,2H),2.62(t,J＝5.3Hz,2H),2.51(dt,J＝14.4,7.3Hz,5H),2.33(t,J＝7.4Hz,6H),2.02(ddt,J＝20.5,16.9,4.3Hz,2H),1.94–1.75(m,5H),1.73–1.44(m,14H),1.43–1.24(m,25H),1.23–0.98(m,8H),0.97–0.82(m,14H),0.71(s,3H).MS-ESI(m/z):785.0(M+H) ⁺ 。

Example 16

The preparation method is the same as compound 1, using 4-bromobutyric acid, 6-amino-1-hexanol, cholesterol, 6-bromohexanoic acid undecyl ester as raw materials to obtain oily compound 15, ¹ H NMR (400MHz, CDCl ₃ )δ4.07(t,J＝6.8Hz,2H),3.65(t,J＝6.6Hz,2H),2.46(dt,J＝11.7,7.2Hz,6H),2.32(td,J＝7.6,1.8Hz,6H),2.08–1.93(m,3H),1.92–1.71(m,6H),1.71–1 .46(m,16H),1.46–1.26(m,28H),1.26–0.85(m,27H),0.70(s,3H).MS-ESI(m/z):841.1(M+H) ⁺ .

Test example

Experimental Example 1 mRNA-LNP Encapsulation and Performance Testing

The mRNA stock solution was dispersed in a 20mM acetic acid solution (pH 5.0) to a final concentration of 200μg/mL (aqueous phase). The mixture was mixed into a lipid mixture (oil phase) according to the molar ratio of Example Compound: Cholesterol: DSPC: DMG-PEG2000 = 50:38.5:10:1.5. The flow rates of the aqueous phase and the oil phase were controlled to mix the mRNA with the lipid mixture by T mixing to obtain the mRNA encapsulated in LNP. The encapsulated LNP was diluted 10 times with a buffer solution, and then concentrated by ultrafiltration, and the diluent was replaced. Finally, the LNP was concentrated to 200ug/mL of mRNA, and the pH of the LNP was adjusted to about 7-8. Finally, the total content and free content of mRNA in the LNP were detected using a Ribogreen kit and 10% OTG as a demulsifier, and the encapsulation rate of the LNP was calculated. The final LNP product was diluted with diluent, 1 ml was added to the particle size pool, and placed on the Malvern ZetaSizer instrument to detect the particle size of the LNP. The results are shown in Table 1.

Particle size, PDI and encapsulation efficiency are important quality attributes of lipid nanoparticles. As shown in the table below, the tested compounds all have good encapsulation efficiency, particle size suitable for mRNA delivery and narrow PDI. LNPs based on compound 3, compound 4, compound 5 and compound 8 have precipitation during the preparation process and poor drugability, so no further studies were conducted.

Table 1: LNP characterization data of example compounds

Compound No. Encapsulation rate/% Particle size/nm PDI 1 97.72 78.76 0.04752 2 97.85 78.35 0.02884 3 98.1 96.34 0.1799 4 63.9 149.8 0.1513 5 89.3 128.9 0.006391 6 85.5 91.39 0.07442 7 79.4 80.68 0.04916 8 92.9 107 0.1525 9 93.6 82.38 0.06221 15 98.9 81.65 0.1541 16 94.1 118.2 0.2082

Test Example 2 Test of intravenous injection delivery effect

1. Encapsulating the mRNA expressing Luciferase into the LNP formulations of compound MC3, compound 1, compound 2, compound 9, compound 15 and compound 16. The LNP formulation preparation method and the mRNA encapsulation method are as described in Experimental Example 1;

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

3. Detect the Luciferase fluorescence expression intensity of each mouse at 6 hours/24 hours.

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

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

Luciferase experiment is the main method to test the expression of mRNA in vivo through fluorescence intensity. According to the results in Figure 1, 6 hours after intravenous injection, compound 15 and compound 16 groups basically did not produce fluorescence, while compound 1, compound 2, and compound 9 groups all produced fluorescence. Compared with the positive control group MC3, the expression of compound 9 group was slightly lower, compound 2 group was comparable, and compound 1 group had significantly higher expression, indicating that it has excellent in vivo expression effect.

As shown in Figure 2, 24 hours after intravenous injection, compared with the positive control group MC3, the compound 1, compound 2, and compound 9 groups had significantly higher expression in the spleen, especially the compound 1 group, which showed that it had excellent spleen targeted delivery effect.

Test Example 3 Test of intramuscular injection delivery effect

1. Encapsulate the mRNA expressing Luciferase into the LNP formulations of compound MC3, compound SM102, compound 1, compound 2, and compound 9. The LNP formulation preparation method and the mRNA encapsulation method are as described in Experimental Example 1;

2. The loaded LNP formulation was injected intramuscularly into Balb/c mice at a dose of 5 μg/mouse, with 5 mice in each group.

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

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

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

As shown in Figure 3, compound 1, compound 2, and compound 9 can also produce fluorescence when administered by intramuscular injection. Among them, the expression of compound 1 and compound 2 are significantly better than MC3, and compound 9 is equivalent to MC3. The expression of compound 1 is better than the positive control group SM102, and compound 2 is only slightly lower than SM102. This reflects that the above compounds also have excellent delivery effects when administered by intramuscular injection.

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

Claims (15)

1. A compound of formula (I), or a pharmaceutically acceptable salt or a stereoisomer thereof, X ₁ and X ₂ are independently selected from C=O or O, and Y ₁ and Y ₂ are independently selected from C=O or O, provided that X ₁ and Y ₁ , X ₂ and Y ₂ are not C=O or O at the same time; _R1 is R; R ₂ and R ₃ are independently selected from H, C1-C14 alkyl, and R, provided that R ₂ and R ₃ are not H at the same time; The R is Ra, Rb, Rc, Rd, Re are independently selected from H, C1-C6 alkyl, R' is selected from C1-C10 alkyl, C1-C10 alkenyl; R ₄ , R ₅ , R ₆ , and R ₇ are independently selected from H, C1-C6 alkyl; Dashed lines represent single or double bonds; n is selected from 0, 1, 2, 3, 4, 5, 6; o and p are independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₂ and R ₃ are independently selected from H, C1-C14 alkyl, provided that R ₂ and R ₃ are not H at the same time, preferably, R ₂ and R ₃ are C6-C11 straight chain or branched chain alkyl, more preferably, R ₂ and R ₃ are C8 straight chain or branched chain alkyl. 3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R' is a C8 alkyl group or a C8 alkenyl group, preferably, R' is The dashed line represents a single bond or a double bond. More preferably, R is: Most preferably, said R is: 4. The compound or pharmaceutically acceptable salt or stereoisomer thereof according to any one of claims 1 to 3, wherein o is selected from 1, 2, 3, 4, 5, 6, 7, preferably, o is 5. 5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein said p is selected from 1, 2, 3, 4, 5, 6, 7, preferably, said p is 7. 6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein n is selected from 2, 3, 4, 5, 6; preferably n is 2 or 6. 7. The compound or pharmaceutically acceptable salt or stereoisomer thereof according to any one of claims 1 to 6, wherein _X1 and _X2 are C=O, and _Y1 and _Y2 are O, or _X1 is O, Y1 is C=O, and _X2 is C=O, and Y2 is O, or _X1 is C=O, _Y1 is O, and _X2 is _O , and Y2 is C=O, or _X1 is C=O, Y1 is O, and X2 is O, and _Y2 is C=O, or _X1 and _X2 are O, and _Y1 and _Y2 are C=O. 8. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, which is a compound of formula (II): 9. A compound or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein the compound is selected from Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 Compound 6 Compound 7 Compound 8 Compound 9 Compound 10 Compound 11 Compound 12 Compound 13 Compound 14 Compound 15 Compound 16 10. A nanoparticle composition comprising a lipid component, wherein the lipid component comprises the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof. 11. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof in the preparation of a lipid nanoparticle composition. 12. A pharmaceutical composition comprising the nanoparticle composition according to any one of claims 10 and a pharmaceutically acceptable carrier. 13. A method for producing a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the nanoparticle composition of any one of claim 10 or the pharmaceutical composition of claim 12 to deliver a therapeutic and/or prophylactic agent to the cell, wherein the therapeutic and/or prophylactic agent is an mRNA encoding the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest. 14. A method for treating a disease or disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of the nanoparticle composition of any one of claim 10 or the pharmaceutical composition of claim 12. 15. A method for specifically delivering a therapeutic and/or prophylactic agent to a mammalian organ, the method comprising administering the nanoparticle composition of any one of claim 10 or the pharmaceutical composition of claim 12 to the mammal, wherein the administration comprises contacting the mammalian organ with the nanoparticle composition, thereby delivering the therapeutic and/or prophylactic agent to the organ.