Granted Patent B2

US 12,239,735 · App. 18/503,327 · Granted Mar 4, 2025

Inventors: Danilo Casimiro (Paris, FR); Sudha Chivukula (Paris, FR); Kirill Kalnin (Paris, FR); Timothy Plitnik (Paris, FR); Timothy Tibbitts (Paris, FR); Frank Derosa (Paris, FR); Anusha Dias (Paris, FR); Rebecca L. Goldman (Paris, FR); Hardip Rajeshbhai Gopani (Paris, FR); Shrirang Karve (Paris, FR); Asad Khanmohammed (Paris, FR); Priyal Patel (Paris, FR)

Assignee: SANOFI

A61K9/1272A61K39/145A61K39/39A61P31/16A61K2039/53A61K2039/55555

Abstract

Claims (70)

1. A pharmaceutical composition comprising a nucleic acid molecule encapsulated in a lipid nanoparticle (LNP), wherein the LNP comprises:

a cationic lipid GL-HEPES-E3-E12-DS-4-E10 at a molar ratio between 35% and 45%,

dimyristoyl-PEG2000 (DMG-PEG2000) at a molar ratio between 0.25% and 2.75%,

cholesterol at a molar ratio between 20% and 35%, and

1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE) at a molar ratio of between 25% and 35%,

wherein all the molar ratios are relative to the total lipid content of the LNP,

wherein the nucleic acid molecule encodes an antigen derived from Propionibacterium acnes.

2. The composition of claim 1 , wherein the LNP comprises:

GL-HEPES-E3-E12-DS-4-E10 at a molar ratio of 40%,

DMG-PEG2000 at a molar ratio of 1.5%,

cholesterol at a molar ratio of 28.5%, and

DOPE at a molar ratio of 30%.

3. The composition of claim 1 , wherein the LNP has an average diameter of 30-200 nm.

4. The composition of claim 1 , wherein the nucleic acid molecule(s) is an mRNA molecule comprising an open reading frame (ORF).

5. The composition of claim 4 , wherein

the LNP comprises two or more mRNA molecules, wherein each mRNA molecule encodes a different Propionibacterium acnes antigen.

6. The composition of claim 4 , wherein

the ORF is codon optimized.

7. The composition of claim 4 , wherein the mRNA molecule comprises at least one 5′ untranslated region (5′ UTR), at least one 3′ untranslated region (3′ UTR), and at least one polyadenylation (poly(A)) sequence.

8. The composition of claim 4 , wherein the mRNA comprises at least one chemical modification.

9. The composition of claim 4 , wherein at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the uracil nucleotides in the mRNA are chemically modified.

10. The composition of claim 8 , wherein the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, 5-methyluridine, 5-methoxyuridine, and 2′-O-methyl uridine.

11. The composition of claim 4 , wherein the composition comprises two or more LNPs, wherein each LNP comprises an mRNA encoding a different Propionibacterium acnes antigen.

12. The composition of claim 1 , wherein the composition is formulated for intramuscular injection.

13. The composition of claim 12 , wherein the composition comprises a phosphate-buffer saline.

14. The composition of claim 12 , wherein the composition comprises trehalose.

15. The composition of claim 14 , wherein the trehalose is at 10% (w/v) of the composition.

16. A kit comprising a container comprising a single-use or multi-use dosage of the composition of claim 1 .

17. The kit of claim 16 , wherein the container is a vial or a pre-filled syringe or injector.

18. A method of preparing the composition of claim 1 , comprising

providing an aqueous buffered solution comprising the nucleic acid molecule,

providing an amphiphilic solution comprising the cationic lipid, the PEGylated lipid, the cholesterol-based lipid, and the helper lipid, and

mixing the aqueous buffered solution and the amphiphilic solution at a ratio of 5:1 to 3:1.

19. The method of claim 18 , comprising mixing the aqueous buffered solution and the amphiphilic solution at a ratio of 4:1.

20. The method of claim 18 , wherein the aqueous buffered solution is an acidic buffered solution.

21. The method of claim 20 , wherein the aqueous buffered solution comprises 1 mM citrate and 150 mM sodium chloride with a pH of about 4.5.

22. The method of claim 18 , wherein the amphiphilic solution is an ethanol solution.

23. A method of eliciting an immune response in a subject in need thereof, comprising administering to the subject a prophylactically effective amount of the composition of claim 1 .

24. The method of claim 23 wherein the composition is administered intramuscularly, intranasally, intravenously, subcutaneously, or intradermally.

25. The method of claim 23 , wherein the method comprises administering to the subject one or more doses of the composition, each dose comprising 1-250 μg of mRNA.

26. The method of claim 23 , wherein the method comprises administering to the subject one or more doses of the composition, each dose comprising 2.5, 5, 15, 45, or 135 μg of mRNA.

27. The method of claim 23 , wherein the method comprises administering to the subject two doses of the composition with an interval of 2-6 weeks.

28. The method of claim 23 , wherein the method comprises administering to the subject two doses of the composition with an interval of 4 weeks.

29. A method of treating or preventing Propionibacterium acnes infections or reducing one or more symptoms of Propionibacterium acnes infections, comprising administering to the subject an effective amount of a pharmaceutical composition comprising a nucleic acid molecule encoding an antigen derived from Propionibacterium acnes encapsulated in a lipid nanoparticle (LNP), wherein the LNP comprises:

a cationic lipid GL-HEPES-E3-E12-DS-4-E10 at a molar ratio between 35% and 45%,

dimyristoyl-PEG2000 (DMG PEG2000) at a molar ratio between 0.25% and 2.75%,

cholesterol at a molar ratio between 20% and 35%, and

1,2-dioleoyl-SN-glycero-3-phosphoethanolamine (DOPE) at a molar ratio of between 25% and 35%,

wherein all the molar ratios are relative to the total lipid content of the LNP.

30. The method of claim 29 , wherein the composition is administered intramuscularly, intranasally, intravenously, subcutaneously, or intradermally.

31. The method of claim 29 , wherein the LNP comprises:

GL-HEPES-E3-E12-DS-4-E10 at a molar ratio of 40%,

DMG-PEG2000 at a molar ratio of 1.5%,

cholesterol at a molar ratio of 28.5%, and

DOPE at a molar ratio of 30%.

32. The method of claim 29 , wherein the LNP has an average diameter of 30-200 nm.

33. The method of claim 29 , wherein the nucleic acid molecule(s) is an mRNA molecule comprising an open reading frame (ORF).

34. The method of claim 29 , wherein

the LNP comprises two or more mRNA molecules, wherein each mRNA molecule encodes a different antigen.

35. The method of claim 33 , wherein the ORF is codon optimized.

36. The method of claim 33 , wherein the mRNA molecule comprises at least one 5′ untranslated region (5′ UTR), at least one 3′ untranslated region (3′ UTR), and at least one polyadenylation (poly(A)) sequence.

37. The method of claim 33 , wherein the mRNA comprises at least one chemical modification.

38. The method of claim 37 , wherein at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the uracil nucleotides in the mRNA are chemically modified.

39. The method of claim 37 , wherein the chemical modification is selected from the group consisting of pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methyluridine, 5-methyluridine, 5-methoxyuridine, and 2′-O-methyl uridine.

40. The method of claim 29 , wherein the LNP has an average diameter of 80-150 nm.

41. The method of claim 29 , wherein the composition comprises 1-10 mg/mL of the LNP.

42. The method of claim 29 , wherein the composition comprises 1 mg/mL of the LNP.

43. The method of claim 29 , wherein the LNP comprises 1-20 nucleic acid molecules.

44. The method of claim 29 , wherein the LNP comprises 5-10 or 6-8 nucleic acid molecules.

45. The method of claim 29 , wherein the composition comprises two or more LNPs, wherein each LNP comprises an mRNA encoding a different antigen.

Assignments (6)

Priority Claims (1)

Continuity (5)

References Cited (329)