Extrusion technique for producing unilamellar vesicles

• US 5,008,050 A
• Filed: 02/13/1989
• Issued: 04/16/1991
• Est. Priority Date: 06/20/1984
• Status: Expired due to Term

• Current Assignee: Cephalon Ltd (Swedish Orphan Biovitrum AB (publ))
• Original Assignee: The Liposome Co., Inc. (Perrigo Company PLC)
• Inventors: Cullis, Pieter R., Hope, Michael J., Bally, Marcel B.
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US07/310,495
• Time in Patent Office: 792 Days
• Field of Search: 264/4.3, 428/402.2, 424/450, 436/829
• US Class Current: 264/4.3
• CPC Class Codes: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: B01D 2317/02 Elements in series
• CPC Class Codes: B01D 61/147 Microfiltration
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

A method for reducing the lamellarity of a population of liposomes is provided which comprises repeatedly passing the liposomes under pressure through a filter which has a pore size equal to or less than about 100 nm. In certain embodiments, the method is used to convert a population of previously formed multilamellar liposomes into a population of substantially unilamellar liposomes. In accordance with other aspects of the disclosure, liposomes are prepared directly from a lipid powder or pellet and buffer without the use of any solvents, detergents or other extraneous materials.

Claims

1. A method for reducing the lamellarity of a population of liposomes comprising repeatedly passing the liposomes under pressure through a filter which has a pore size (equal to or less than) no larger than about 100 nm.

2. The method of claim 1 wherein the population of liposomes is passed through the filter more than two times.

3. The method of claim 1 wherein the population of liposomes is passed through the filter at a pressure greater than 100 pounds per square inch.

4. The method of claim 3 wherein the flow rate through the filter is greater than or equal to about 20 milliliters per minute.

5. The method of claim 1 wherein the filter has straight through channels.

6. The method of claim 5 wherein the filter is a polycarbonate filter.

7. The method of claim 6 wherein the filter comprises two or more polycarbonate filters arranged in series.

8. The method of claim 1 wherein the population of liposomes has a lipid concentration greater than 60 umol/ml.

9. The method of claim 1 wherein the population of liposomes is passed through the filter more than two times at a pressure greater than 100 pounds per square inch and wherein the filter has straight through channels.

10. A method for preparing a population of substantially unilamellar liposomes comprising the steps of:

(a) preparing multilamellar liposomes; and

(b) repeatedly passing the liposomes under pressure through a filter which has a pore size no larger than about 100 nm.

11. The method of claim 10 wherein the liposomes are passed through the filter more than two times.

12. The method of claim 10 wherein the liposomes are passed through the filter at a pressure greater than 100 pounds per square inch.

13. The method of claim 10 wherein the flow rate through the filter is greater than or equal to about 20 milliliters per minute.

14. The method of claim 10 wherein the filter has straight through channels.

15. The method of claim 14 wherein the filter is a polycarbonate filter.

16. The method of claim 15 wherein the filter comprises two or more polycarbonate filters arranged in series.

17. The method of claim 10 wherein the population of liposomes has a lipid concentration greater than 60 umol/ml.

18. The method of claim 10 wherein the multilamellar liposomes are prepared by mechanical agitation of an aqueous suspension of one or more lipids.

19. The method of claim 10 including the additional step of subjecting the liposomes to a freeze-thaw cycle to increase their trapped volumes.

20. The method of claim 10 wherein the multilamellar liposomes are prepared by mechanical agitation of an aqueous suspension of one or more lipids, wherein the liposomes are passed through the filter more than two times at a pressure greater than 100 pounds per square inch, and wherein the filter has straight through channels.

21. The method of claim 20 including the additional step of subjecting the liposomes to a freeze-thaw cycle to increase their trapped volumes.

22. A method for preparing liposomes without the use of solvents, detergents or other extraneous materials comprising the steps of:

(a) preparing a mixture of a lipid powder or pellet and an aqueous buffer; and

(b) repeatedly passing the mixture under pressure through a filter, wherein the filter has a pore size less than or equal to about 100 nm, the mixture is passed through the filter more than two times, and the resulting liposomes are substantially unilamellar.

23. The method of claim 22 wherein the mixture is passed through the filter at a pressure greater than 100 pounds per square inch.

24. The method of claim 23 wherein the flow rate through the filter is greater than or equal to about 20 milliliters per minute.

25. The method of claim 23 wherein the filter has straight through channels.

26. The method of claim 25 wherein the filter is a polycarbonate filter.

27. The method of claim 26 wherein the filter comprises two or more polycarbonate filters arranged in series.

28. The method of claim 22 wherein the mixture has a lipid concentration greater than 60 umol/ml.

29. The method of claim 22 including the additional step of subjecting the liposomes to a freeze-thaw cycle to increase their trapped volumes.