Reconstitutable dry powder formulations and methods of use thereof

Abstract

The present disclosure is directed to the use of reconstituted mRNA dry powder particles for parenteral administration. The present disclosure is also directed to a method of generating dry powder particles supplemented with appropriate excipients for optimal thermostability and in vivo expression.

Description

Abstract This disclosure relates to the use of reconstituted mRNA dry powder particles for parenteral administration. This disclosure also relates to a method for generating dry powder particles, wherein suitable excipients are added to the dry powder particles to achieve optimal thermostability and in vivo expression.

Claims

We claim:

1. A dry powder formulation for reconstitution comprising:(a) an excipient that is sucrose or trehalose; and(b) a lipid nanoparticle (LNP) comprising a messenger RNA (mRNA) encapsulated by one or more lipids; wherein the excipient of (a) and total lipids in the LNP of (b) have a weight ratio of at least about 5.

2. The dry powder formulation of claim 1, wherein the excipient is sucrose.

3. The dry powder formulation of claim 1, wherein the excipient is trehalose.

4. The dry powder formulation of any one of claims 1-3, wherein the weight ratio of the excipient to the total lipids in the LNP is at least about 5.6, at least about 11, or at least about 15.

5. The dry powder formulation of claim 4, wherein the weight ratio of the excipient to the total lipids is at least about 11.1 or at least about 15.6.

6. The dry powder formulation of any one of claims 1-5, wherein the LNP comprises one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids.

7. A dry powder formulation for reconstitution comprising(a) an excipient that is a sugar or sugar alcohol; and(b) a lipid nanoparticle (LNP) comprising a messenger RNA (mRNA) encapsulated by one or more lipids; wherein weight ratio of the excipient of (a) to total lipids in the LNP of (b) is at least about 5.

8. The dry powder formulation of claim 7, wherein the sugar or sugar alcohol is sucrose, mannitol, xylitol, lactose, or trehalose.

9. The dry powder formulation of claim 7, wherein the sugar or sugar alcohol is trehalose.

10. The dry powder formulation of claim 7, wherein the sugar or sugar alcohol is sucrose.

11. The dry powder formulation of any one of claims 7-10, wherein the weight ratio of the sugar or sugar alcohol to the total lipids in the LNP is at least about 5.6, at least about11. or at least about 15.

12. The dry powder formulation of claim 7, wherein the weight ratio of the sugar or sugar alcohol to the total lipids in the LNP is at least about 11.1 or at least about 15.6.

13. The dry powder formulation of any one of claims 7-12, wherein the LNP comprises one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids.

14. The dry powder formulation of any one of claims 1-13, wherein the particle size of the reconstituted dry powder formulation is less than about 120 nm.

15. The dry powder formulation of any one of claims 1-14, wherein the encapsulation rate is greater than about 60%.

16. The dry powder formulation of any one of claims 1-15, wherein the particle size of the dry powder is less than about 5pm.

17. The dry powder formulation of any one of claims 1-16, wherein the dry powder formulation is stable after prolonged storage, for example, after prolonged storage at 2-8 °C (e.g., at 4°C).

18. The dry powder formulation of claim 17, wherein the mRNA maintains an integrity of about 80% or greater after storage, e.g., after storage at 2-8°C (e.g., 4°C) for at least about six months.

19. The dry powder formulation of claim 18, wherein the mRNA maintains an integrity of about 80% or greater, e.g., after storage at 2-8°C (e.g., 4°C) for up to about 1 year.

20. The dry powder formulation of claim 19, wherein the mRNA maintains an integrity of about 80% or greater, optionally where mRNA integrity is measured after storage at 2- 8°C (e.g., 4°C) for at least 1 year.

21. The dry powder formulation of any one of claims 1-20, wherein the mRNA encodes a therapeutic protein.

22. The dry powder formulation of any one of claims 1-21, wherein the mRNA encodes an antigen.

23. The dry powder formulation of any one of claims 1-22, wherein the formulation is suitable for a vaccine.

24. The dry powder formulation of any one of claims 1-23, wherein the formulation is suitable for parenteral delivery once reconstituted.

25. The dry powder formulation of claim 24, wherein the formulation is suitable for intramuscular, intravenous, or subcutaneous delivery once reconstituted.

26. The dry powder formulation of any one of claims 1-25, wherein the formulation is suitable for mucosal delivery once reconstituted.

27. The dry powder formulation of claim 26, wherein the formulation is suitable for oral, sublingual, or intranasal delivery once reconstituted.

28. A method of delivering a mRNA in vivo, the method comprising administering to a subject in need thereof a reconstituted form of the dry powder formulation of any one of claims 1-27.

29. A method of treating a disease or disorder in a subject, the method comprising administering to the subject a reconstituted form of the dry powder formulation of any one of claims 1-27.

30. The method of claim 28 or 29, wherein the reconstituted form of the dry powder formulation is administered subcutaneously, intravenously, or intramuscularly.

31. A method of preparing a dry powder formulation for reconstitution, the method comprising:(a) combining a first mixture comprising lipid nanoparticles (LNPs) and an ethanolic solution, thereby obtaining a second mixture, wherein the LNPs comprise a messenger RNA (mRNA) encapsulated by one or more lipids, and wherein the ethanolic solution comprises an excipient that is sucrose or trehalose at a concentration of about 1- 11% w / v, and(b) spray-drying the second mixture, thereby obtaining the dry powder formulation.

32. A method of preparing a reconstituted dry powder formulation, the method comprising:(a) providing a dry powder formulation prepared according to the method of claim 31 ; and(b) reconstituting the dry powder formulation in water or buffer, thereby obtaining the reconstituted dry powder formulation.

33. The method of claim 32, wherein the LNPs in the reconstituted dry powder formulation have a diameter of about 100 nm.

34. The method of any one of claims 31-33, wherein sucrose or trehalose and total lipids in the LNPs have a weight ratio of at least about 5, at least about 11, or at least about 15.

35. The method of any one of claims 31-34, wherein the concentration of sucrose or trehalose is greater than about 2% (w / v), greater than about 5% (w / v), or greater than about 7% (w / v) post-reconstitution.

36. The method of any one of claims 31-35, wherein the concentration of sucrose or trehalose is from about 2% (w / v) to about 10% (w / v) post-reconstitution.

37. The method of any one of claims 32-36, wherein the particle size of the reconstituted dry-powder formulation is less than about 120 nm.

38. The method of any one of claims 32—37, wherein the reconstituted LNP particle size is between about 80 nm and about 120 nm.

39. The method of any one of claims 32-38, wherein the reconstituted LNP particle size is between about 80 nm and about 115 nm.

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