Single vial vaccine formulations

1. A thermostable lyophilized vaccine composition comprising:

a metabolizable oil;

a cake-forming excipient, wherein the cake-forming excipient is (1) a combination of mannitol and trehalose or (2) a trehalose; and

an adjuvant, wherein the adjuvant is a TLR4 agonist,

wherein the composition is formed by lyophilization of an oil-in-water emulsion formulation, does not comprise glycerol, does not contain liposomes, does not contain alum, is in the form of a cake, forms an oil-in-water emulsion upon reconstitution, and is thermostable at a temperature between about 8° C. to about 60° C. for at least 10 months.

2. The composition of claim 1 , wherein the cake-forming excipient is trehalose which is at a concentration of about 10% (w/v) in the oil-in-water emulsion formulation.

3. The composition of claim 1 , wherein the cake-forming excipient is trehalose which is at a concentration of about 5% (w/v) in the oil-in-water emulsion formulation.

4. The composition of claim 1 , wherein the composition is formed by lyophilization of an oil-in water emulsion formulation, wherein the cake-forming excipient is a combination of mannitol and trehalose, wherein the mannitol in the oil-in water emulsion formulation is at a concentration of about 0.1% (w/v) and trehalose in the oil-in water emulsion formulation is at a concentration of about 5% (w/v).

5. The composition of claim 1 , wherein the composition is formed by lyophilization of an oil-in water emulsion formulation, wherein the cake-forming excipient is a combination of mannitol and trehalose, and wherein the mannitol in the oil-in water emulsion formulation is at a concentration of about 2.5% (w/v) and trehalose in the oil-in water emulsion formulation is at a concentration of about 2.5% (w/v).

6. The composition of claim 1 wherein the composition is thermostable for at least 12 months.

7. The composition of claim 1 wherein the composition is thermostable at about 25° C. for at least 10 months.

8. The composition of claim 1 wherein the composition is thermostable at about 37° C. for at least 10 months.

9. The composition of claim 1 wherein the composition is thermostable at about 50° C. for at least 10 months.

10. The composition of claim 1 wherein the composition is in the form of an elegant cake.

11. The composition of claim 1 wherein the cake does not exhibit browning by visual inspection when stored at temperature between about 8° C. to about 60° C. for at least 1 month.

12. The composition of claim 1 , wherein the thermostability of the composition is determined prior to reconstitution of the composition.

13. The composition of claim 12 , wherein the cake does not exhibit shrinking, cracking and/or browning.

14. The composition of claim 1 , wherein the thermostability is determined following reconstitution of the composition.

15. The composition of claim 14 , wherein thermostability is determined by inspection of the oil-in-water emulsion formed upon reconstitution for creaming.

16. The composition of claim 1 , wherein thermostability is determined by assay of the components of the oil-in-water emulsion formed upon reconstitution.

17. The composition of claim 1 , wherein the oil-in-water emulsion formed upon reconstitution has a particle size with a Z-average diameter of less than about 200 nm.

18. The composition of claim 1 , further comprising an antigen.

19. The composition of claim 18 , wherein the antigen is a polypeptide, a nucleic acid encoding a polypeptide, or a pathogen.

20. The composition of claim 1 , wherein the metabolizable oil is squalene, synthetic squalene, grape seed oil, olive oil, or a synthetic isoprenoid.

21. The composition of claim 1 , wherein the TLR4 agonist is MPL, 3d-MPL, or synthetic GLA.

22. The composition of claim 21 , wherein the synthetic GLA has the following structure:

wherein R 1 , R 3 , R 5 , and R 6 are C 11 -C 20 alkyl; and R 2 and R 4 are C 9 -C 20 alkyl.

23. The composition of claim 22 , wherein R 1 , R 3 , R 5 and R 6 are C 11 alkyl; and R 2 and R 4 are C 9 alkyl.

24. The composition of claim 1 , wherein the metabolizable oil is squalene, synthetic squalene, grape seed oil, olive oil or a synthetic isoprenoid.

25. The composition of claim 1 , further comprising 1,2-dimyristoy 1-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycerol-3-phsphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), egg phosphatidylcholine (PC), lecithin, a polysorbate surfactant, or a combination thereof.

26. The composition of claim 1 , further comprising a surfactant.

27. The composition of claim 26 , wherein the surfactant is poloxomer 188.

28. The composition of claim 1 , further comprising an antioxidant.

29. The composition of claim 28 , wherein the antioxidant is vitamin E.

30. A single vial comprising the thermostable lyophilized vaccine of claim 1 , wherein the composition is contained in the vial.

31. A method of storing the thermostable lyophilized vaccine composition of claim 1 at between about 25° C. to about 60° C. for at least 10 months.

32. A method for generating the thermostable lyophilized vaccine composition of claim 1 , comprising the step of lyophilizing an oil-in-water emulsion to form the thermostable lyophilized vaccine composition, wherein the oil-in-water emulsion prior to lyophilization comprises the metabolizable oil and the cake-forming excipient.

33. A method of stimulating an immune response in a subject comprising: (a) reconstituting the thermostable lyophilized vaccine composition of claim 1 into an oil-in-water emulsion, wherein the composition is in the form of a cake and (b) administering the emulsion to the subject, thereby stimulating an immune response in the subject.

34. The composition of claim 1 , and wherein the adjuvant concentration in the oil-in-water emulsion formed upon reconstitution exhibits no more than about 25% breakdown of the adjuvant concentration in the oil-in-water emulsion formulation prior to lyophilization.

35. The composition of claim 18 , wherein the antigen concentration in the oil-in-water emulsion formed upon reconstitution exhibits no more than about 25% breakdown of the antigen concentration in the oil-in-water emulsion formulation prior to lyophilization.

36. A thermostable lyophilized vaccine composition comprising:

a metabolizable oil;

a cake-forming excipient, wherein the cake-forming excipient is (1) a combination of mannitol trehalose or (2) trehalose; and

an adjuvant, wherein the adjuvant is a TLR4 agonist,

wherein the composition is formed by lyophilization of an oil-in-water emulsion formulation, does not comprise glycerol, does not contain liposomes, is in the form of a cake, forms an oil-in-water emulsion upon reconstitution, and is thermostable at a temperature of about 50° C. for at least 10 months.

37. The composition of claim 36 , wherein the cake-forming excipient is trehalose which is at a concentration of about 10% (w/v) in the oil-in-water emulsion formulation.

38. The composition of claim 36 , wherein the cake-forming excipient is trehalose which is at a concentration of about 5% (w/v) in the oil-in-water emulsion formulation.

39. The composition of claim 36 , wherein the composition is formed by lyophilization of an oil-in water emulsion formulation, wherein the cake-forming excipient is a combination of mannitol and trehalose, wherein the mannitol in the oil-in water emulsion formulation is at a concentration of about 0.1% (w/v) and trehalose in the oil-in water emulsion formulation is at a concentration of about 5% (w/v).

40. The composition of claim 36 , wherein the composition is formed by lyophilization of an oil-in water emulsion formulation, wherein the cake-forming excipient is a combination of mannitol and trehalose, and wherein the mannitol in the oil-in water emulsion formulation is at a concentration of about 2.5% (w/v) and trehalose in the oil-in water emulsion formulation is at a concentration of about 2.5% (w/v).

41. The composition of claim 36 , wherein the cake does not exhibit browning by visual inspection after storage.

42. A thermostable lyophilized vaccine composition comprising:

a metabolizable oil, wherein the metabolizable oil is squalene;

1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC);

a cake-forming excipient, wherein the cake-forming excipient is (1) a combination of mannitol and trehalose or (2) trehalose; and

an adjuvant, wherein the adjuvant is glucopyranosyl lipid adjuvant (GLA),

wherein the composition is in the form of a cake, is formed by lyophilization of an oil-in-water emulsion formulation, and the oil-in-water emulsion formulation does not comprise glycerol, wherein the composition is thermostable at a temperature between 8° C. to 60° C. for at least 10 months.

43. The composition of claim 42 , wherein the composition is thermostable at a temperature of about 50° C. for at least 10 months.

44. The composition of claim 42 , wherein the cake-forming excipient comprises at least about 5% (w/v) trehalose.