Liposomal vaccine

• US 20050169979A1
• Filed: 01/14/2005
• Published: 08/04/2005
• Est. Priority Date: 07/03/2002
• Status: Abandoned Application

• Current Assignee: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. (Hebrew University of Jerusalem), Receptor BioLogix, Inc.
• Original Assignee: Receptor BioLogix, Inc.
• Inventors: Hagan, Susan Anne, Barenholz, Yechezkel, Michaeli, Dov, Grimes, Stephen, Even-Chen, Simcha
• Application Number:
  US11/036,690
• Publication Number:
  US 20050169979A1
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 38/09 Luteinising hormone-releasing hormone [LHRH] , i.e. Gonadotropin-releasing hormone [GnRH]; Related peptides
• CPC Class Codes: A61K 38/2207 Gastrins; Cholecystokinins [CCK]
• CPC Class Codes: A61K 38/27 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: A61K 9/127 Liposomes

Abstract

The invention provides liposomal vehicles for encapsulating relatively high levels of immunogenic protein substances including immunogens directed against hormones and hormone receptors, such as gastrin and gonadotropin releasing hormone and their receptors. The liposome encapsulating large amounts of immunogens can be injected parenterally to induce effective immune responses without exhibiting significant adverse tissue reactogenicity. Methods for production of the liposomal vaccines and methods of their administration for treatment of diseases and conditions associated with the cognate hormones are also provided.

Claims

1. A liposomal vaccine formulation comprising an immunogenic protein substance and a liposome-forming phospholipid in an ethanolic saline comprising from about 1% to about 10% ethanol by volume.

2. The liposomal vaccine formulation according to claim 1, wherein the ethanolic saline comprises about 5% ethanol by volume.

3. The liposomal vaccine formulation according to claim 1, wherein the ethanolic saline substantially eliminates foaming of the vaccine formulation during mixing.

4. The liposomal vaccine formulation according to claim 1, wherein the weight ratio of phospholipid to immunogenic protein substance is between about 50:

1 and about 1000;

1.

5. The liposomal vaccine formulation according to claim 4, wherein the weight ratio of phospholipid to immunogenic protein substance is about 300:

1.

6. The liposomal vaccine formulation according to claim 1, wherein the immunogenic protein substance is at least about 65% encapsulated within liposomes.

7. The liposomal vaccine formulation according to claim 1, wherein the immunogenic protein substance is at least about 80% associated with liposomes.

8. The liposomal vaccine formulation according to claim 1, wherein the liposome-forming phospholipid comprises one or more of the following:

phosphatidic acid (PA), phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl inositol (PI), dimyristoyl phosphatidyl choline (DMPC) and dimyristoyl phosphatidyl glycerol (DMPG).

9. The liposomal vaccine formulation according to claim 8, wherein the liposome-forming phospholipid is dimyristoyl phosphatidyl choline (DMPC).

10. The liposomal vaccine formulation according to claim 1, comprising multilamellar vesicles (MLVs).

11. The liposomal vaccine formulation according to claim 1, wherein the immunogenic protein substance comprises an immunomimic peptide of a hormone or a hormone receptor.

12. The liposomal vaccine formulation according to claim 11, wherein the immunomimic peptide is peptide having the sequence of gastrin G17, gastrin G34, GnRH, hCG, or fragments thereof.

13. The liposomal vaccine formulation according to claim 12, wherein the peptide sequence is the gastrin G17 of SEQ ID NO:

1.

14. The liposomal vaccine formulation according to claim 12, wherein the peptide sequence is a gastrin G17 fragment sequence selected from the group consisting of SEQ ID NOS:

3-8.

15. The liposomal vaccine formulation according to claim 12, wherein the peptide sequence is the gastrin G34 of SEQ ID NO:

12.

16. The liposomal vaccine formulation according to claim 12, wherein the peptide sequence is GnRH immunomimic peptide of SEQ ID NO:

15.

17. The liposomal vaccine formulation according to claim 12, wherein the peptide sequence is the hCG immunomimic peptide of SEQ ID NO:

16.

18. The liposomal vaccine formulation according to claim 12, wherein the immunomimic peptide is conjugated to an immunogenic carrier through a spacer peptide.

19. The liposomal vaccine formulation according to claim 18, wherein the spacer peptide is selected from the group consisting of SEQ ID NOS:

9-11.

20. The liposomal vaccine formulation according to claim 1, further comprising a cytokine a muramyl-dipeptide or a murametide.

21. The liposomal vaccine formulation according to claim 1, which is a sterile injectable formulation.

22. The liposomal vaccine formulation according to claim 1, further comprising an excipient that facilitates hydration of the formulation, the excipient comprising one or more of:

(i) from about 0.01% to about 10% by weight of a saccharide;

(ii) from about 0.01% to about 10% by weight of a tricarboxylic acid; and

(iii) a buffer at a pH from about 5.0 to about 9.0.

23. The liposomal vaccine formulation according to claim 22, wherein the saccharide is sucrose.

24. The liposomal vaccine formulation according to claim 22, wherein the tricarboxylic acid is citric acid.

25. The liposomal vaccine formulation according to claim 22, wherein the buffer is a phosphate buffer, a citrate buffer or a bicarbonate buffer.

26. The liposomal vaccine formulation according to claim 22, wherein the buffer has a pH of from about 6.0 to 8.0.

27. The liposomal vaccine formulation according to claim 22, wherein the buffer has a pH of about 7.

28. The liposomal vaccine formulation according to claim 1, wherein the vaccine is in a dose of from about 50 μ

g to about 5 mg.

29. A method of treatment of a gastrointestinal disease or disorder comprising administering to a patient in need thereof an effective amount of a liposomal vaccine formulation comprising an immunomimic peptide having the sequence of gastrin G17, gastrin G34, or fragments thereof, and a liposome-forming phospholipid in an ethanolic saline comprising from about 1% to about 10% ethanol by volume.

30. The method of treatment according to claim 29, wherein the ethanolic saline comprises about 5% ethanol.

31. The method of treatment according to claim 29, wherein the weight ratio of phospholipid to immunogenic protein substance is between about 50:

1 and about 1000;

1.

32. The method of treatment according to claim 31, wherein the weight ratio of phospholipid to immunogenic protein substance is about 300:

1.

33. The method of treatment according to claim 29, wherein the immunomimic peptide is at least about 65% encapsulated within the liposomes.

34. The method of treatment according to claim 29, wherein the immunomimic peptide is at least about 80% associated with liposomes.

35. The method of treatment according to claim 29, wherein the phospholipid comprises one or more of the following:

phosphatidic acid (PA), phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl inositol (PI), dimyristoyl phosphatidyl choline (DMPC) and dimyristoyl phosphatidyl glycerol (DMPG).

36. The method of treatment according to claim 35, wherein the phospholipid is dimyristoyl phosphatidyl choline (DMPC).

37. The method of treatment according to claim 29, wherein the liposomal vaccine formulation comprises multilamellar vesicles (MLVs).

38. A method of preparing a liposomal vaccine formulation comprising:

(a) providing a liposome-forming phospholipid in an organic solvent and an aqueous solution comprising an immunogenic protein substance, wherein the weight ratio of liposome-forming phospholipid to immunogenic protein substance is between about 50;

1 and about 1000;

1;

(b) mixing the organic solvent containing the liposome-forming phospholipid and the aqueous solution comprising the immunogenic protein substance to form an emulsion;

(c) removing the organic solvent from to form a gel-like mixture; and

(d) hydrating the gel-like mixture with ethanolic saline comprising from about 1% to about 10% ethanol by volume.

39. The method according to claim 38, wherein the ethanolic saline comprises about 5% ethanol by volume.

40. The method according to claim 38, wherein the weight ratio of phospholipid to immunogenic protein substance is about 300:

1.

41. The method according to claim 38, wherein the immunogenic protein substance is at least about 65% encapsulated within liposomes.

42. The method according to claim 38, wherein the immunogenic protein substance is at least about 80% associated with liposomes.

43. A pharmaceutical composition comprising a liposomal vaccine formulation comprising an immunogenic protein substance and a liposome-forming phospholipid in an ethanolic saline comprising from about 1% to about 10% ethanol by volume.