Gas and oxygen carrying lipid vesicles

• US 5,219,538 A
• Filed: 03/01/1991
• Issued: 06/15/1993
• Est. Priority Date: 03/13/1987
• Status: Expired due to Term

• Current Assignee: Novavax Inc.
• Original Assignee: Micro-Pak, Inc.
• Inventors: Henderson, Sheryl L., Mathur, Rajiv, Wallach, Donald F. H.
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US07/662,850
• Time in Patent Office: 837 Days
• Field of Search: 264/4.1, 264/4.3, 264/4.32, 264/4.33, 264/4.6, 264/4.7, 428/402.2, 424/450, 436/829, 514/832
• US Class Current: 428/402.2
• CPC Class Codes: A61K 49/227 Liposomes, lipoprotein vesicles, e.g. LDL or HDL lipoproteins, micelles, e.g. phospholipidic or polymeric
• CPC Class Codes: A61K 8/14 Liposomes; Vesicles
• CPC Class Codes: A61K 8/22 Peroxides; Oxygen; Ozone
• CPC Class Codes: A61K 8/70 containing perfluoro groups, e.g. perfluoroethers
• CPC Class Codes: A61K 9/0026 Blood substitute; Oxygen transporting formulations; Plasma extender
• 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: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: A61K 9/1278 Post-loading, e.g. by ion or pH gradient
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: B01J 13/203 Exchange of core-forming material by diffusion through the capsule wall
• CPC Class Codes: C11D 17/0021 Aqueous microemulsions
• CPC Class Codes: C11D 17/0026 Structured liquid compositions, e.g. liquid crystalline phases or network containing non-Newtonian phase
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

The present invention relates to the production of lipid vesicles having a moderately volatile material such as a perfluorocarbon or a silicone oil encapsulated therein. In another aspect, lipid vesicles having a gas-filled center or core are made. The lipid vesicles having gas-filled central core are made by dehydrating vesicles having the moderately volatile liquid encapsulated in the central core. This drives off the water first, allowing the moderately volatile liquid to stabilize the vesicle structure as it dries, finally forming a central void which can refract light. The preferred vesicles of the invention are paucilamellar vesicles.

Claims

1. A paucilamellar lipid vesicle comprising a large, amorphous central cavity surrounded by 2-10 lipid bilayers having a moderately volatile liquid encapsulated in said central cavity, said moderately volatile liquid having the following properties:

a) it is less volatile than aqueous solutions,

b) it is more volatile than the lipid and any materials forming said lipid bilayers,

c) it can be evaporated from said lipid vesicle without damaging the structural integrity of said lipid vesicle,

d) it is substantially immiscible with aqueous solutions,

e) it is substantially immiscible with the lipid forming said lipid bilayers, and

f) it is substantial unreactive with the lipid in said lipid bilayer.

2. The paucilamellar lipid vesicle of claim 1 wherein said moderately volatile liquid comprises a perfluorocarbon.

3. An oxygen carrier comprising the perfluorocarbon filled vesicle of claim 2.

4. The paucilamellar vesicle of claim 2 wherein said perfluorocarbon is selected from the group consisting of perfluorodecalin, perfluorohexane, perfluorooctane, perfluorodimethylcyclohexane, perfluorotripropylamine, octafluorocyclobutane, and mixtures thereof.

5. The paucilamellar lipid vesicle of claim 4 wherein said perfluorocarbon comprises a perfluorocarbon capable of carrying oxygen.

6. The paucilamellar lipid vesicle of claim 1 further comprising an oil, said oil being immiscible with said lipid, immiscible with said aqueous solutions and immiscible with said moderately volatile liquid.

7. The paucilamellar lipid vesicle of claim 6 wherein said oil has oil-soluble materials carried therein.

8. The paucilamellar lipid vesicle of claim 1 wherein said lipid comprises a lipid with a melting temperature of at least about 40°

C.

9. The paucilamellar lipid vesicle of claim 8 wherein said lipid comprises a nonionic lipid.

10. The paucilamellar lipid vesicle of claim 1 wherein said lipid comprises a phospholipid.

11. A method of manufacturing gas-filled paucilamellar lipid vesicles having 2-10 lipid bilayers surrounding a substantially amorphous, gas-filled central cavity comprising the steps of:

a) hydrating the lipid to be used as the primary structural wall material with an aqueous solution to form paucilamellar lipid vesicles filled with said aqueous solution in said central cavity,

b) mixing said lipid vesicles with a moderately volatile liquid that is (1) substantially immiscible with said lipid, (2) substantially immiscible with aqueous solutions, (3) less volatile than aqueous solutions, (4) more volatile than said lipid, (5) substantially unreactive with lipid, and (6) can be evaporated from said lipid without damaging the structural integrity of said paucilamellar lipid vesicles to form moderately volatile liquid filled paucilamellar lipid vesicles,

c) removing said aqueous solution and said moderately volatile material from said paucilamellar lipid vesicles without damaging the structural integrity of said paucilamellar lipid vesicles,

whereby said gas-filled paucilamellar vesicles are formed.

12. The method of claim 11 wherein said removing step comprises air drying.

13. The method of claim 11 wherein said removing step comprises vacuum drying.

14. The method of claim 11 wherein said removing step comprises lyophilizing.

15. The method of claim 11 wherein said moderately volatile liquid comprises a perfluorocarbon.

16. The method of claim 15 wherein said perfluorocarbon is selected from the group consisting of perfluorodecalin, perfluorohexane, perfluorooctane, perfluorodimethylcyclohexane, perfluorotripropylamine, octafluorocyclobutane, and mixtures thereof.

17. The method of claim 11 wherein said lipid comprises a lipid with a melting temperature of at least about 40°

C.

18. The method of claim 11 wherein said mixing step further comprises mixing oil that is immiscible with said lipid and immiscible in said moderately volatile liquid.

19. The method of claim 11 comprising the step of dispersing said moderately volatile liquid in said lipid before hydration, thereby simultaneously carrying out said steps of hydrating said lipid and mixing said lipid vesicles with said moderately volatile liquid.