Method and composition for the site-selective delivery using viruses encapsulated in microbubbles

• US 8,454,937 B2
• Filed: 08/22/2007
• Issued: 06/04/2013
• Est. Priority Date: 09/12/2006
• Status: Active Grant

• Current Assignee: Temple University of The Commonwealth System of Higher Education
• Original Assignee: Temple University of The Commonwealth System of Higher Education
• Inventors: Claudio, Pier Paolo
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US11/894,821
• Publication Number:
  US 20080063604A1
• Time in Patent Office: 2,113 Days
• Field of Search: None
• US Class Current: 424/9.52
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A61K 41/0028 Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
• CPC Class Codes: A61K 47/6901 Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors
• CPC Class Codes: A61K 9/0009 involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis microelectromechanical systems A61K9/0097
• CPC Class Codes: A61K 9/5015 Organic compounds, e.g. fats, sugars
• CPC Class Codes: A61K 9/5036 Polysaccharides, e.g. gums, alginate; Cyclodextrin
• CPC Class Codes: A61K 9/5052 Proteins, e.g. albumin

Abstract

Methods and compositions for the site-selective delivery of a gene to a site within the body of an animal where virus not encapsulated within the microbubbles is inactivated, for example by treatment with a virus-inactivating agent.

Claims

1. A method for preparing a composition for the site-selective delivery of a virus to a site within the body of an animal, the method comprising:

(1) providing a composition comprising a microbubble, wherein the microbubble comprises

(a) a gas,

(b) a shell surrounding the gas, and

(c) a virus encapsulated within the microbubble; and

(2) treating the composition with a virus-inactivating agent to render virus not encapsulated in the microbubble inactive.

3. The method according to claim 1, wherein the average microbubble diameter is not more than about 10 μ

m.

4. The method according to claim 3, wherein the average microbubble diameter is not more than about 5 μ

M.

5. The method according to claim 1, wherein the microbubble shell comprises a phospholipid.

6. The method according to claim 5, wherein the phospholipid is selected from the group consisting of phosphatidylcholines, phosphatidylglycerols, and phosphatidylserines.

7. The method according to claim 6, wherein the phospholipid is dimyristoylphosphatidylcholine.

8. The method according to claim 1, wherein the microbubble shell comprises a protein.

9. The method according to claim 8, wherein the protein is albumin.

10. The method according to claim 1, wherein the gas comprises nitrogen.

11. The method according to claim 1, wherein the gas comprises at least one fluorocarbon.

12. The method according to claim 11, wherein the gas comprises at least one perfluoralkane.

13. The method according to claim 12, wherein the gas comprises a perfluoroalkane selected from the group consisting of perfluorobutane and perfluorohexane.

14. The method according to claim 1, wherein the virus comprises a viral vector comprising an expression cassette encoding a molecule of interest.

15. The method according to claim 14, wherein the molecule of interest is a therapeutic molecule.

16. The method according to claim 14, wherein the molecule of interest is selected from the group consisting of green fluorescent protein, luciferase, chloramphenicol transferase, β

-glucuronidase, and β

-galactosidase.

17. The method according to claim 1, wherein the virus is selected from the group consisting of adenoviruses, parvoviruses, retroviruses, herpesviruses, poxviruses, vaccinia viruses, and baculoviruses.

18. The method according to claim 17, wherein the virus is an adenovirus.

19. The method according to claim 1, wherein the virus-inactivating agent comprises a chemical composition.

20. The method according to claim 19, wherein the virus-inactivating agent is complement.

44. The method according to claim 15, wherein the therapeutic molecule is a nucleic acid.

45. The method according to claim 44, wherein the nucleic acid is selected from the group consisting of antisense RNA, antisense DNA, ribozymes, and siRNA.

2. A method for the site-selective delivery of a virus to a specific site within the body of an animal, the method comprising:

(1) providing a composition comprising a microbubble, wherein the microbubble comprises

(a) a gas,

(b) a shell surrounding the gas, and

(c) a virus encapsulated within the microbubble;

(2) treating the composition with a virus-inactivating agent to render virus not encapsulated in the microbubble inactive;

(3) administering the composition to the animal; and

(4) directing ultrasound waves to the specific site within the body of the animal to release the virus from the microbubble.

21. A composition for the site-selective delivery of a virus to a site within the body of an animal, the composition formed by a process comprising:

(1) providing a composition comprising a microbubble, wherein the microbubble comprises

(a) a gas,

(b) a shell surrounding the gas, and

(c) a virus encapsulated within the microbubble; and

(2) treating the composition with a virus-inactivating agent to render virus not encapsulated in the microbubble inactive.

22. The composition according to claim 21, wherein the average microbubble diameter is not more than about 10 μ

m.

23. The composition according to claim 21, wherein the average microbubble diameter is not more than about 5 μ

m.

24. The composition according to claim 21, wherein the microbubble shell comprises a phospholipid.

25. The composition according to claim 24, wherein the phospholipid is selected from the group consisting of phosphatidylcholines, phosphatidylglycerols, and phosphatidylserines.

26. The composition according to claim 24, wherein the phospholipid is dimyristoylphosphatidylcholine.

27. The composition according to claim 21, wherein the microbubble shell comprises a protein.

28. The composition according to claim 27, wherein the protein is albumin.

29. The composition according to claim 21, wherein the gas comprises nitrogen.

30. The composition according to claim 21, wherein the gas comprises at least one fluorocarbon.

31. The composition according to claim 30, wherein the gas comprises at least one perfluoralkane.

32. The composition according to claim 31, wherein the gas comprises a perfluoroalkane selected from the group consisting of perfluorobutane and perfluorohexane.

33. The composition according to claim 21, wherein the virus comprises a viral vector comprising an expression cassette encoding a molecule of interest.

34. The composition according to claim 33, wherein the molecule of interest is a therapeutic molecule.

35. The composition according to claim 33, wherein the molecule of interest is selected from the group consisting of green fluorescent protein, luciferase, chloramphenicol transferase, β

-glucuronidase, and β

-galactosidase.

36. The composition according to claim 21, wherein the virus is selected from the group consisting of adenoviruses, parvoviruses, retroviruses, herpesviruses, poxviruses, vaccinia viruses, and baculoviruses.

37. The composition according to claim 36, wherein the virus is an adenovirus.

38. The composition according to claim 21, wherein the virus-inactivating agent comprises a chemical composition.

39. The composition according to claim 38, wherein the virus-inactivating agent is complement.

40. The composition according to claim 21, wherein the composition is in the form of an aqueous solution.

46. The composition according to claim 34, wherein the therapeutic molecule is a nucleic acid.

47. The composition according to claim 46, wherein the nucleic acid is selected from the group consisting of antisense RNA, antisense DNA, ribozymes, and siRNA.

41. A composition for the site-selective delivery of a virus to a site within the body of an animal, the composition comprising a microbubble, wherein the microbubble comprises:

(a) a gas,

(b) a shell surrounding the gas, and

(c) a virus encapsulated within the microbubble; and

(d) inactive virus that is not encapsulated within the microbubble.

42. A method for the site-selective delivery of a virus to a specific site within the body of an animal, the method comprising:

(1) administering to the animal a composition comprising a microbubble, wherein the microbubble comprises;

(a) a gas,

(b) a shell surrounding the gas, and

(c) a virus encapsulated within the microbubble; and

(d) inactive virus that is not encapsulated within the microbubble; and

(2) directing ultrasound waves to the specific site within the body of the animal to release the virus from the microbubble.

43. A kit for preparing a composition for the site-selective delivery of a virus to a site within the body of an animal, the kit comprising:

(1) in a first compartment, a composition comprising a microbubble, wherein the microbubble comprises

(a) a gas, and

(b) a shell surrounding the gas;

(2) in a second compartment, a virus-inactivating agent; and

(3) instructional material comprising instructions for preparing a composition of microbubbles comprising encapsulated virus for site-selective delivery of the virus.