Stable lipid-comprising drug delivery complexes and methods for their production

• US 20050152964A1
• Filed: 12/15/2004
• Published: 07/14/2005
• Est. Priority Date: 01/23/1995
• Status: Active Grant

• Current Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Original Assignee: University Of Pittsburgh (University of Pittsburgh of The Commonwealth System of Higher Education)
• Inventors: Sloane, David L., Loomis, Aaron Garth, Paul, Ralph Wilfred, Huang, Leaf, Gao, Xiang, Sorgi, Frank L.
• Granted Patent: US 7,335,509 B2
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

Novel stable, concentrated, biologically active and ready-to-use lipid-comprising drug delivery complexes and methods for their production are described. The biological activity of the complexes produced are comparable to the formulations prepared according to the prior art admixture method and upon purification, the complexes produced by the method of this invention are 50 to 500 fold more concentrated than the complexes formed by admixture. The method described herein provides for the large scale production of lipid-comprising drug delivery systems useful for gene therapy and other applications.

Claims

1. (canceled)

2. (canceled)

3. (canceled)

4. (canceled)

5. (canceled)

6. (canceled)

7. (canceled)

8. (canceled)

9. (canceled)

10. (canceled)

11. (canceled)

12. A method for producing drug/lipid/polycation complexes having a positive charge excess of lipid and polycation to drug, said method comprising:

mixing said drug with cationic liposomes and at least one polycation in a ratio of drug to lipid to polycation such that said complexes are formed.

16. The method of claim 12, wherein the cationic liposome comprises a cationic lipid and a neutral phospholipid.

17. The method of claim 16, wherein the cationic lipid is DC-Chol.

18. The method of claim 17, wherein the neutral phospholipid is dioleoyl phosphatidylethanolamine.

19. The method of claim 18, wherein the drug is nucleic acid.

20. The method of claim 12, wherein said complex has an average diameter less than 300 nm.

21. The method of claim 20, wherein the average diameter of said formulation remains substantially unchanged for up to one year in storage.

13. (canceled)

14. (canceled)

15. (canceled)

22. A drug/lipid/polycationic polypeptide complex comprising drug, at least one lipid species, and at least one polycationic polypeptide in a ratio such that said complex has a positive charge excess of lipid and polycationic polypeptide to drug.

23. The complex of claim 22, wherein said polycationic polypeptide has an amino acid composition in which arginine residues constitute greater than 30% of the amino acid residues of the polypeptide and lysine residues constitute less than about 5% of the amino acid residues of the polypeptide.

24. The complex of claim 23, wherein said drug is a nucleic acid.

25. The complex of claim 24, wherein said complex has a diameter of less than about 400 nm.

26. The complex of claim 24, wherein said complex has a drug/cationic peptide ratio between about 1:

0.01 and 1;

100.

27. The complex of claim 24, wherein said polypeptide from about 20 to about 100 amino acids in length.

40. A method for delivering drug to cells comprising:

contacting said cells with the complex of claim 22.

41. The method of claim 40, wherein the drug to be delivered is a nucleic acid molecule which encodes a protein or peptide.

43. The method of claim 40, wherein the cells are contacted with the complex in vivo, said method comprising administering the complex to an animal or human in an amount effective to deliver the drug into the cells of the animal or the human.

28. (canceled)

29. A method for producing drug/lipid/polycationic polypeptide complexes, said method comprising mixing drug to lipid to polycationic polypeptide in a ratio of from about 1 μ

g/0.1 nmol/0.01 μ

g to about 1 μ

g/200 nmol/100 μ

g.

31. The method of claim 29, wherein the polycationic polypeptide has an amino acid composition in which the arginine residues constitute greater than about 30% of the amino acid residues of the polypeptide and the lysine residues constitute less than about 5% of the amino acid residues of the polypeptide.

32. The method of claim 31, wherein the drug is a nucleic acid molecule.

33. The method of claim 32, wherein the lipid species is a cationic liposome.

30. (canceled)

34. (canceled)

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)

42. (canceled)

44. (canceled)

45. (canceled)