Liposomal defensins

• US 5,766,624 A
• Filed: 05/24/1995
• Issued: 06/16/1998
• Est. Priority Date: 10/25/1993
• Status: Expired due to Fees

• Current Assignee: Transave Inc. (Insmed Incorporated)
• Original Assignee: Liposme Company Incorporated
• Inventors: Perkins, Walter, Ahmad, Imran, Janoff, Andrew S.
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US08/449,598
• Time in Patent Office: 1,119 Days
• Field of Search: 424/450, 424/417, 514/2, 514/21
• US Class Current: 424/450
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/04 Immunostimulants

Abstract

Defensins are microbicidal, tumoricidal cytotoxic protein components of animal host defense systems. They can be entrapped in liposomes containing release-inhibiting lipid such that the defensins are neutralized, and their release from the liposomes is inhibited; however, the defensins remain effective when exposed to endocytosed material in endocytic vesicles. Liposomal defensin formulations are administered to animals for the treatment or prevention of microbial infections, for the treatment of cancers and for the treatment of disorders characterized by a deficiency of protein-mediated cytotoxic activity in cytoplasmic granules.

Claims

1. A method of treating a mammal for an infection by a microbe sensitive to a defensin, the method comprising administering to the mammal a liposome comprising:

(a) a bilayer comprising a lipid which comprises palmitoyloleoyl phosphatidylcholine, dioleoyl phosphatidylcholine or distearoyl phosphatidylcholine and cholesterol;

(b) an internal compartment comprising an aqueous buffer; and

,

(c) a neutralized defensin selected from the group consisting of prototypical mammalian defensins, beta-defensins, indolicidin, magainins and insect defensins,

wherein an amount of the liposome comprising an anti-infection effective amount of the defensin is administered.

2. The method of claim 1, wherein the mammal is an immunocompromised human.

3. The method of claim 1, wherein the microbe is a fungus.

4. The method of claim 1, wherein the mammal is an immunocompromised human and the infection is by a fungus selected from the group consisting of Aspergilli and Cryptococci.

5. The method of claim 1, wherein the anti-infection effective amount of the defensin is between about 1 and about 1000 mg of the defensin per kg of the mammal'"'"'s body weight.

9. The method of claim 1 or 6, wherein the liposome has multiple bilayers.

10. The multilamellar liposome of claim 9 having a solute entrapped in its internal compartments, wherein the concentration of solute in each of the compartments is substantially equal.

11. The method of claim 1 or 6, wherein the defensin is indolicidin.

12. The method of claim 1 or 6, wherein the defensin is associated with the bilayer.

13. The method of claim 1 or 6, wherein the defensin is entrapped in an internal compartment of the liposome.

14. The method of claim 1 or 6, wherein the defensin comprises at most about 20 mole percent of the liposome.

15. The method of claim 1 or 6, wherein the aqueous buffer is a citrate buffer.

16. The method of claim 1 or 6, wherein the lipid further comprises an ionizable cationic lipid.

17. The liposome of claim 16, wherein the lipid further comprises about 5 mole percent of the lipid 1,2-dipalmitoyl-3-(N,N-dimethylamino)-propane.

18. The method of claim 1 or 6 wherein the lipid further comprises a headgroup-modified lipid.

19. The method of claim 1 or 6, wherein the liposome comprises an additional bioactive agent selected from the group consisting of antiparasitic, antifungal, antibacterial or antineoplastic compounds.

20. The method of claim 1 or 6, wherein the lipid comprises dioleoyl phosphatidylcholine, wherein the liposome comprises a citrate buffer and about 0.5 mole percent of the defensin indolicidin, and wherein the liposome is a multilamellar liposome having substantially equal interlamellar solute distribution.

21. The method of claim 1 or 6, wherein the lipid comprises distearoyl phosphatidylcholine and cholesterol, wherein the liposome comprises a citrate buffer and about 0.5 mole percent of the defensin indolicidin, and wherein the liposome is a multilamellar liposome having substantially equal interlamellar solute distribution.

22. The method of claim 1 or 6, wherein the liposome is dehydrated and subsequently rehydrated prior to administration.

6. A method of treating a mammal afflicted with a cancer responsive to a defensin, the method comprising administering to the mammal a liposome comprising:

(a) a bilayer comprising a lipid which comprises palmitoyloleoyl phosphatidylcholine, dioleoyl phosphatidylcholine or distearoyl phosphatidylcholine and cholesterol;

(b) an internal compartment comprising an aqueous buffer; and

, (c) a neutralized defensin selected from the group consisting of prototypical mammalian defensins, beta-defensins, indolicidin, magainins and insect defensins,

wherein an amount of the liposome comprising an anticancer effective amount of the defensin is administered to the mammal.

7. The method of claim 6, wherein the cancer is a leukemia or lymphoma.

8. The method of claim 6, wherein the anticancer effective amount of the defensin is between about 1 and about 1000 mg of the defensin per kg of the mammal'"'"'s body weight.