Gas filled liposomes and stabilized gas bubbles and their use as ultrasonic contrast agents

• US 5,769,080 A
• Filed: 02/22/1994
• Issued: 06/23/1998
• Est. Priority Date: 12/22/1989
• Status: Expired due to Term

• Current Assignee: Lantheus Medical Imaging, Inc. (Lantheus Holdings, Inc.)
• Original Assignee: The Dupont Merck Pharmaceutical Company
• Inventors: Unger, Evan C., Wu, Guanli
• Primary Examiner(s): JAWORSKI, FRANCIS J
• Application Number:
  US08/199,462
• Time in Patent Office: 1,582 Days
• Field of Search: 128/660.01, 128/662.02, 424/9, 264/4.1, 264/4.3, 422/436
• US Class Current: 600/458
• 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 49/1815 compo-inhalant, e.g. breath tests
• 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: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2982 Particulate matter [e.g., sphere, flake, etc.]

Abstract

Contrast agents for ultrasonic imaging comprising gas filled liposomes prepared using vacuum drying gas instillation methods, and gas filled liposomes substantially devoid of liquid in the interior thereof, are described. Methods of and apparatus for preparing such liposomes and methods for employing such liposomes in ultrasonic imaging applications are also disclosed. Also described are diagnostic kits for ultrasonic imaging which include the subject contrast agents.

Claims

1. A gas filled liposome prepared by a vacuum drying gas instillation method.

2. A gas filled liposome of claim 1 wherein said liposome is comprised of lipid materials selected from the group consisting of fatty acids, lysolipids, dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidic acid, sphingomyelin, cholesterol, cholesterol hemisuccinate, tocopherol hemisuccinate, phosphatidylethanolamine, phosphatidylinositol, lysolipids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids, and polymerized lipids.

3. A gas filled liposome of claim 2 wherein said liposome is comprised of dipalmitoylphosphatidylcholine.

4. A gas filled liposome of claim 2 wherein said liposome comprises polymerized lipids.

5. A gas filled liposome of claim 2 further comprising polyethyleneglycol.

6. A gas filled liposome of claim 2 wherein said liposome is filled with a gas selected from the group consisting of air, nitrogen, carbon dioxide, oxygen, argon, xenon, helium, and neon.

7. A gas filled liposome of claim 6 wherein said liposome is filled with nitrogen gas.

8. A gas filled liposome of claim 1 wherein said liposome is stored suspended in an aqueous medium.

9. A gas filled liposome of claim 1 wherein said liposome is stored dry.

10. A gas filled liposome of claim 1 wherein said liposome has a stability of greater than about three weeks.

11. A gas filled liposome of claim 1 wherein said liposome has a reflectivity of greater than about 2 dB.

12. A gas filled liposome of claim 11 wherein said liposome has a reflectivity of between about 2 dB and about 20 dB.

13. A gas filled liposome of claim 1 wherein said liposome comprises a material for in vivo targeting.

14. A gas filled liposome of claim 13 wherein said targeting material comprises a carbohydrate.

15. A gas filled liposome of claim 1 wherein said liposome is stable to changes in pressure.

16. A gas filled liposome of claim 1 wherein said liposome is selected from the group consisting of oligolamellar liposomes and unilamellar liposomes.

17. A gas filled liposome of claim 16 wherein said liposome comprises oligolamellar liposomes.

18. A gas filled liposome of claim 16 wherein said liposome comprises unilamellar liposomes.

19. A gas filled liposome of claim 18 which comprises a phospholipid.

20. A gas filled liposome of claim 16 which comprises a phospholipid.

21. A gas filled liposome substantially devoid of liquid in the interior thereof.

22. A gas filled liposome of claim 21 wherein said liposome is comprised of lipid materials selected from the group consisting of fatty acids, lysolipids, dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidic acid, sphingomyelin, cholesterol, cholesterol hemisuccinate, tocopherol hemisuccinate, phosphatidylethanolamine, phosphatidylinositol, lysolipids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids, and polymerized lipids.

23. A gas filled liposome of claim 22 wherein said liposome is comprised of dipalmitoylphosphatidylcholine.

24. A gas filled liposome of claim 22 wherein said liposome comprises polymerize lipids.

25. A gas filled liposome of claim 22 further comprising polyethyleneglycol.

26. A gas filled liposome of claim 21 wherein said liposome is filled with a gas selected from the group consisting of air, nitrogen, carbon dioxide, oxygen, argon, xenon, helium, and neon.

27. A gas filled liposome of claim 26 wherein said liposome is filled with nitrogen gas.

28. A gas filled liposome of claim 21 wherein said liposome is stored suspended in an aqueous medium.

29. A gas filled liposome of claim 21 wherein said liposome is stored dry.

30. A gas filled liposome of claim 21 wherein said liposome has a shelf life stability of greater than about three weeks.

31. A gas filled liposome of claim 21 wherein said liposome has a reflectivity of greater than about 2 dB.

32. A gas filled liposome of claim 31 wherein said liposome has a reflectivity of between about 2 dB and about 20 dB.

33. A gas filled liposome of claim 21 wherein said liposome comprises a material for in vivo targeting.

34. A gas filled liposome of claim 33 wherein said material comprises a carbohydrate.

35. A gas filled liposome of claim 21 wherein said liposome is stable to changes in pressure.

36. A gas filled liposome of claim 21 wherein said liposome is selected from the group consisting of oligolamellar liposomes and unilamellar liposomes.

37. A gas filled liposome of claim 36 wherein said liposome comprises oligolamellar liposomes.

38. A gas filled liposome of claim 36 wherein said liposome comprises unilamellar liposomes.

39. A gas filled liposome of claim 38 which comprises a phospholipid.

40. A gas filled liposome of claim 36 which comprises a phospholipid.

41. A contrast agent adapted to be inject ed in to the body of a patient and comprising stabilized gas bubbles which comprises gas encapsulated by one or more lipid materials, said stabilized gas bubbles being substantially devoid of liquid in the interior thereof.

42. A contrast agent of claim 41 wherein said stabilized gas bubbles comprise liposomes that are selected from the group consisting of oligolamellar liposomes and unilamellar liposomes.

43. A contrast agent of claim 42 wherein said stabilized gas bubbles comprise oligolamellar liposomes.

44. A contrast agent of claim 42 wherein said stabilized gas bubbles comprise unilamellar liposomes.

45. A contrast agent of claim 44 wherein said lipid materials comprise phospholipids.

46. A contrast agent of claim 42 wherein said lipid materials comprise phospholipids.

47. A contrast agent of claim 41 wherein said lipid materials are selected from the group consisting of fatty acids, lysolipids, dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidic acid, sphingomyelin, cholesterol, cholesterol hemisuccinate, tocopherol hemisuccinate, phosphatidylethanolamine, phosphatidylinositol, lysolipids, sphingomyelin, glycosphingolipids, glucolipids, glycolipids, sulphatides, lipids with ether and ester-linked fatty acids, and polymerized lipids.

48. A contrast agent of claim 47 wherein said lipid materials comprise dipalmitoylphosphatidylcholine.

49. A contrast agent of claim 47 wherein said lipid materials comprise polymerized lipids.

50. A contrast agent of claim 41 further comprising polyethyleneglycol.

51. A contrast agent of claim 41 which comprises a material for in vivo targeting.

52. A contrast agent of claim 51 wherein said material comprises a carbohydrate.

53. A contrast agent of claim 41 wherein said gas is selected from the group consisting of air, nitrogen, carbon dioxide, oxygen, argon, xenon, helium, and neon.

54. A contrast agent of claim 41 wherein said stabilized gas bubbles are stored suspended in an aqueous medium.

55. A contrast agent of claim 41 which is stored dry.

56. A contrast agent of claim 41 wherein said stabilized gas bubbles are stable to changes in pressure.