Contrasting agents for ultrasonic imaging and methods for preparing the same

• US 6,001,335 A
• Filed: 06/18/1996
• Issued: 12/14/1999
• Est. Priority Date: 12/22/1989
• Status: Expired due to Term

• Current Assignee: Lantheus Medical Imaging, Inc. (Lantheus Holdings, Inc.)
• Original Assignee: Imarx Pharmaceutical Corp. (Sycamore Entertainment Group, Inc.)
• Inventors: Unger, Evan C.
• Primary Examiner(s): Dees, Jose' G.
• Assistant Examiner(s): Hartley, Michael G.
• Application Number:
  US08/665,719
• Time in Patent Office: 1,274 Days
• Field of Search: 424/9.51, 424/9.52, 424/450, 424/489, 424/490, 424/498, 424/502, 128/662.02, 600/458, 600/441, 264/4, 264/4.1, 264/4.3, 264/4.32, 427/2.14, 427/213.3, 428/402.2
• US Class Current: 424/9.52
• CPC Class Codes: A61K 41/0028 Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
• CPC Class Codes: A61K 41/0052 Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
• CPC Class Codes: A61K 49/223 Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
• 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 9/127 Liposomes
• 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: Y10T 428/2984 Microcapsule with fluid core [includes liposome]

Abstract

Liposomes suitable as ultrasound contrast agents which contain media of various types including gases, gaseous precursors activated by pH, temperature or pressure, as well as other solid or liquid contrast enhancing agents, are described. Methods of using the same as ultrasound contrast agents are also disclosed. The present invention also comprises novel methods for synthesizing liposomes having encapsulated therein gases.

Claims

1. A method for synthesizing a lipid vesicle for use in ultrasound imaging, the lipid vesicle having encapsulated therein a gas, wherein the method comprises subjecting a solvent-free composition which comprises a gaseous precursor and a lipid in an aqueous medium to a first pressure and thereafter subjecting said composition to a second pressure, wherein said second pressure is a decreased pressure.

2. A method according to claim 1 wherein said lipid in said composition which is subjected to decreased pressure is in the form of a lipid vesicle, said lipid vesicle containing said gaseous precursor.

3. A method according to claim 1 wherein said lipid vesicles comprise one or more lipids selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine and phosphatidic acid.

4. A method according to claim 1 wherein said lipid vesicles further comprise a polymer on the surface of said lipid vesicles.

5. A method according to claim 4 wherein said polymer comprises polyethylene glycol.

6. A method according to claim 3 wherein said lipid vesicles further comprise a polymer on the surface of said lipid vesicles.

7. A method according to claim 6 wherein said polymer comprises polyethylene glycol.

8. A method according to claim 1 wherein said lipid vesicles comprise liposomes.

9. A method according to claim 1 wherein said lipid vesicles comprise one or more phospholipids.

10. A method according to claim 9 wherein said phospholipid is selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, lysolipids, fatty acids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids and polymerizable lipids.

11. A method according to claim 10 wherein said phospholipid is a polymerizable lipid.

12. A method according to claim 1 wherein said lipid vesicles further comprise polyethyleneglycol.

13. A method according to claim 1 further comprising placing said composition under decreased temperature.

14. A method according to claim 13 wherein said decreased temperature is from about 1 to about 4°

C.

15. A method according to claim 8 wherein said liposomes are selected from the group consisting of unilamellar liposomes, oligolamellar liposomes and multilamellar liposomes.

16. A method according to claim 15 wherein said liposomes comprise unilamellar liposomes.

17. A method according to claim 16 wherein said liposomes comprise one or more phospholipids.

18. A method according to claim 17 wherein said phospholipid is selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, lysolipids, fatty acids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids and polymerizable lipids.

19. A method according to claim 18 wherein said phospholipid is a polymerizable lipid.

20. A method according to claim 18 wherein said liposomes further comprise polyethyleneglycol.

21. A method according to claim 15 wherein said liposomes are selected from the group consisting of oligolamellar liposomes and multilamellar liposomes.

22. A method according to claim 21 wherein said liposomes comprise one or more phospholipids.

23. A method according to claim 22 wherein said phospholipid is selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, lysolipids, fatty acids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids and polymerizable lipids.

24. A method according to claim 23 wherein said phospholipid is a polymerizable lipid.

25. A method according to claim 23 wherein said liposomes further comprise polyethyleneglycol.

26. A method according to claim 13 wherein said composition is placed under decreased temperature at about the time said composition is subjected to said first pressure.

27. A method for synthesizing a lipid vesicle having encapsulated therein a gas wherein the method comprises contacting vesicles in an aqueous medium with gas under first and second pressures, wherein said second pressure is a decreased pressure.

28. A method according to claim 27 wherein said lipid vesicle comprises a liposome.

29. A method according to claim 28 wherein said liposome is selected from the group consisting of unilamellar liposomes, oligolamellar liposomes and multilamellar liposomes.

30. A method according to claim 28 wherein said liposome is a unilamellar liposome.

31. A method according to claim 30 wherein said lipid is a phospholipid.

32. A method according to claim 30 wherein said phospholipid is selected from the group consisting of cholesterol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, lysolipids, fatty acids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids and polymerizable lipids.

33. A method according to claim 32 wherein said phospholipid is a polymerizable lipid.

34. A method according to claim 32 wherein said liposomes further comprise polyethyleneglycol.

35. A method according to claim 29 wherein said liposomes are selected from the group consisting of oligolamellar and multilamellar liposomes.

36. A method according to claim 35 wherein said lipid is a phospholipid.

37. A method according to claim 27 wherein said gas is selected from the group consisting of nitrogen, oxygen, carbon dioxide, xenon, argon, neon and helium.

38. A method according to claim 27 further comprising placing said vesicles under decreased temperature.

39. A method according to claim 38 wherein said decreased temperature is from about 1 to about 4°

C.

40. A method according to claim 38 wherein said vesicles are placed under decreased temperature at about the time the vesicles are subjected to said first pressure.

41. A method for synthesizing a lipid vesicle for use in ultrasound imaging, the lipid vesicle having encapsulated therein a gas, the method consisting essentially of subjecting a composition which comprises a gaseous precursor and a lipid in an aqueous medium to a first pressure and thereafter subjecting said composition to a second pressure, wherein said second pressure is a decreased pressure.