Fibrinogen-coated liposomes

• US 5,658,588 A
• Filed: 03/31/1995
• Issued: 08/19/1997
• Est. Priority Date: 03/31/1995
• Status: Expired due to Term

• Current Assignee: University of Cincinnati
• Original Assignee: University of Cincinnati
• Inventors: Deanglis, Ashley P., Retzinger, Gregory Scott
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US08/414,368
• Time in Patent Office: 872 Days
• Field of Search: 424/450, 436/829, 264/4.1, 264/4.3, 514/2, 514/21
• US Class Current: 424/450
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911

Abstract

A method for preparing fibrinogen-coated liposomes is disclosed. In this process, fibrinogen and an acylating agent are reacted in the presence of a dispersion of liposomes under specifically defined reaction conditions. The liposomes formed using this process, pharmaceutical compositions containing those liposomes, and the methods of clotting blood and delivering pharmaceutically-active agents and/or other chemicals utilizing those pharmaceutical compositions are also disclosed.

Claims

1. A liposome wherein a hemostatically effective amount of a C₈ -C₂₀ acylated fibrinogen is covalently incorporated into the lipid bilayer of said liposome.

2. A liposome according to claim 1 which is formed from phospholipid materials selected from the group consisting of phosphatidylcholine, phosphatidic acid, lysophosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, sphingolipids, phosphatidylglycerol, sphingomyelin, cardiolipin, glycolipids, gangliosides, cerebrosides, and mixtures thereof.

3. A liposome according to claim 2 having a size of from about 0.05 to about 20 microns.

4. A liposome according to claim 3 having a saturated or subsaturated film of acylated fibrinogen on the liposome surface.

5. A liposome according to claim 4 wherein the acylated fibrinogen is a C₁₂ -C₁₈ acylated fibrinogen.

6. A liposome according to claim 5 wherein from about 1% to about 95% of the ε

-amino functional sites of the lysines of the fibrinogen are acylated.

7. A liposome according to claim 6 wherein the acylated fibrinogen is a C₁₆ acylated fibrinogen.

8. A liposome according to claim 7 having a size of from about 0.2 to about 1 micron.

9. A liposome according to claim 8 having a size of about 0.4 micron.

10. A liposome according to claim 5 containing from about 0.1 to about 1.0 milligram of acylated fibrinogen per milligram of liposome.

11. A liposome according to claim 8 containing from about 0.6 to about 0.7 mg. of acylated fibrinogen per milligram of liposome.

12. A liposome according to claim 11 wherein from about 20% to about 25% of the ε

-amino functional sites of the lysines of fibrinogen are acylated.

13. A liposome according to claim 6 which additionally comprises a safe and effective amount of a pharmaceutically-active agent incorporated within said liposome.

14. A liposome according to claim 13 wherein said pharmaceutically active agent is selected from the group consisting of anti-inflammatory agents, clot dissolving agents, radiodiagnostic agents, and mixtures thereof.

15. A liposome according to claim 6 which additionally comprises a safe and effective amount of nucleic acids incorporated within said liposome.

16. A pharmaceutical composition comprising a safe and effective amount of the liposome according to claim 6, and the balance of said composition being a pharmaceutically-acceptable compatible carrier.

17. A pharmaceutical composition according to claim 16 which comprises from about 0.001% to about 98% of said liposome.

18. A pharmaceutical composition comprising a safe and effective amount of the liposome according to claim 11, and the balance of said composition being a pharmaceutically-acceptable compatible carrier.

19. A pharmaceutical composition according to claim 18 which comprises from about 0.001% to about 98% of said liposome.

20. A pharmaceutical composition comprising a safe and effective mount of the liposome according to claim 14, and the balance being a pharmaceutically acceptable compatible carrier.

21. A pharmaceutical composition according to claim 20 which comprises from about 0.001% to about 98% of said liposome.

35. A method for providing hemostasis at a wound site comprising administering a safe and effective amount of the pharmaceutical composition according to claim 17 to said wound site together with a safe and catalytically effective amount of thrombin.

36. A method for delivering a pharmaceutically active agent to the site of accumulation of fibrin or fibrinogen comprising the administration of a safe and effective mount of a pharmaceutical composition according to claim 21 to a patient in need of such treatment.

22. A method of preparing fibrinogen-coated liposomes comprising reacting fibrinogen and an acylating agent in the presence of a dispersion of liposomes at a buffered pH of from about 6.5 to about 8.5 and a temperature of from about 0°

C. to about 50°

C., wherein the molar ratio of said acylating agent to said fibrinogen is from about 1;

1 to about 5,000;

1, and there is sufficient fibrinogen in the reaction to form a film on the surface of said liposomes.

23. A method according to claim 22 wherein the liposomes are formed from a phospholipid material.

24. A method according to claim 23 wherein the acylating agent is selected from the group consisting of C₈ -C₂₀ acid chlorides, C₈ -C₂₀ acid anhydrides and mixtures thereof.

25. A method according to claim 24 wherein the acylating agent is a C₈ -C₂₀ acid chloride.

26. A method according to claim 25 wherein the size of said liposomes is from about 0.05 to about 20 microns.

27. A method according to claim 26 wherein the molar ratio of acylating agent to fibrinogen is from about 2,500:

1 to about 5,000;

1.

28. A method according to claim 27 wherein the acylating agent is a C₁₂ -C₁₈ acid chloride.

29. A method according to claim 28 wherein the liposomes have a size of from about 0.2 to about 1 micron.

30. A method according to claim 29 wherein the acylating agent is palmitoyl chloride.

31. A method according to claim 30 wherein the molar ratio of acylating agent to fibrinogen is about 2,500:

1.

32. A method according to claim 31 wherein the size of the liposomes is about 0.4 microns.

33. The liposomes made according to the process of claim 22.

34. The liposomes made according to the process of claim 32.