Bilayer stabilizing components and their use in forming programmable fusogenic liposomes

• US 5,885,613 A
• Filed: 06/07/1995
• Issued: 03/23/1999
• Est. Priority Date: 09/30/1994
• Status: Expired due to Fees

• Current Assignee: University of British Columbia
• Original Assignee: University of British Columbia
• Inventors: Cullis, Pieter R., Madden, Thomas D., Holland, John W.
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US08/485,608
• Time in Patent Office: 1,385 Days
• Field of Search: 426/450, 428/402.2
• 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: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

The present invention provides a fusogenic liposome comprising a lipid capable of adopting a non-lamellar phase, yet capable of assuming a bilayer structure in the presence of a bilayer stabilizing component; and a bilayer stabilizing component reversibly associated with the lipid to stabilize the lipid in a bilayer structure. Such fusogenic liposomes are extremely advantageous because the rate at which they become fusogenic can be not only predetermined, but varied as required over a time scale ranging from minutes to days. Control of liposome fusion can be achieved by modulating the chemical stability and/or exchangeability of the bilayer stabilizing component(s). The fusogenic liposomes of the present invention can be used to deliver drugs, peptide, proteins, RNA, DNA or other bioactive molecules to the target cells of interest.

Claims

1. A fusogenic liposome comprising:

a lipid capable of adopting a non-lamellar phase, yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate, wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines, phosphatidylserines, ceramides, glycolipids and mixtures thereof; and

a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure, wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0.05 mole percent to about 50 mole percent.

2. The fusogenic liposome in accordance with claim 1 wherein said lipid is a phosphatidylethanolamine.

3. The fusogenic liposome in accordance with claim 2 wherein said lipid is an unsaturated phosphatidylethanolamine.

4. The fusogenic liposome in accordance with claim 1 wherein said lipid is a mixture of a phosphatidylethanolamine and a phosphatidylserine.

5. The fusogenic liposome in accordance with claim 1 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid.

6. The fusogenic liposome in accordance with claim 5 wherein said cationic lipid is a member selected from the group consisting of 3β

-(N-(N'"'"',N'"'"'-dimethylaminoethane)carbamoyl)cholesterol, N,N-distearyl-N,N-dimethylammonium bromide, N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide, diheptadecylamidoglycyl spermidine, N-(1-(2,3-dioleyloxy)propyl)-N-(2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate, N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride, N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride and N,N-dioleyl-N,N-dimethylammonium chloride.

7. The fusogenic liposome in accordance with claim 1 further comprising cholesterol.

8. The fusogenic liposome in accordance with claim 1 wherein said polyethyleneglycol has a molecular weight ranging from about 200 to 10,000.

9. The fusogenic liposome in accordance with claim 1 wherein said polyethyleneglycol has a molecular weight ranging from about 1,000 to 8,000.

10. The fusogenic liposome in accordance with claim 1 wherein said polyethyleneglycol has a molecular weight ranging from about 2,000 to 6,000.

11. The fusogenic liposome in accordance with claim 1 wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0.05 mole percent to about 30 mole percent.

12. The fusogenic liposome in accordance with claim 1 wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 1 mole percent to about 20 mole percent.

13. The fusogenic liposome in accordance with claim 7 wherein said cholesterol is present at a concentration ranging from about 0.02 mole percent to about 50 mole percent.

14. The fusogenic liposome in accordance with claim 1 wherein the rate at which said liposome becomes fusogenic can be varied over a timescale ranging from minutes to days.

15. The fusogenic liposome in accordance with claim 1 wherein the rate at which said liposome becomes fusogenic is controlled by controlling the composition of said polyethyleneglycol-ceramide conjugate in said fusogenic liposome.

16. The fusogenic liposome in accordance with claim 1 wherein the rate at which said liposome becomes fusogenic is controlled by controlling the concentration of said polyethyleneglycol-ceramide conjugate in said fusogenic liposome.

17. A method for delivering a therapeutic compound to a target cell at a predetermined rate, comprising;

administering to a host containing said target cell a fusogenic liposome which comprises a polyethyleneglycol-ceramide conjugate, a lipid capable of adopting a non-lamellar phase, yet capable of assuming a bilayer structure in the presence of said polyethyleneglycol-ceramide conjugate and said therapeutic compound or a pharmaceutically acceptable salt thereof, wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines, phosphatidylserines, ceramides, glycolipids and mixtures thereof, and wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0.05 male percent to about 50 mole percent.

18. The method in accordance with claim 17 wherein said lipid is a phosphatidylethanolamine.

19. The method in accordance with claim 17 wherein said lipid is a mixture of an phosphatidylethanolamine and a phosphatidylserine.

20. The method in accordance with claim 17 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid.

21. The method in accordance with claim 17 wherein said fusogenic liposome further comprises cholesterol.

22. The method in accordance with claim 17 wherein said fusogenic liposome is administered intravenously.

23. The method in accordance with claim 17 wherein said fusogenic liposome is administered parenterally.

24. The method in accordance with claim 17 wherein said fusogenic liposome administered to said host is unilamellar.

25. The method in accordance with claim 24 wherein said unilamellar fusogenic liposome has a mean diameter of 0.05 microns to 0.45 microns.

26. The method in accordance with claim 25 wherein said unilamellar fusogenic liposome has a mean diameter of 0.05 microns to 0.2 microns.

27. A method of stabilizing in a bilayer structure a lipid which is capable of adopting a non-lamellar phase, said method comprising:

combining a polyethylene glycol-ceramide conjugate with a lipid which is capable of adopting a non-lamellar phase, yet which is capable of assuming a bilayer structure in the presence of said polyethylene glycol-ceramide conjugate, said polyethylene glycol-ceramide conjugate being selected to be exchangeable or biodegradable such that over a predetermined period of time, said polyethylene glycol-ceramide conjugate is lost from said bilayer structure or modified, thereby rendering said bilayer structure fusogenic wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines, phosphatidylserines, ceramides, glycolipids and mixtures thereof, and wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0.5 mole percent to about 50 mole percent.

28. The method in accordance with claim 27 wherein said lipid is a phosphatidylethanolamine.

29. The method in accordance with claim 27 wherein said lipid is a mixture of a phosphatidylethanolamine and a phosphatidylserine.

30. The method in accordance with claim 27 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid.

31. The method in accordance with claim 27 further comprising cholesterol.