Liposomes as contrast agents for ultrasonic imaging and methods for preparing the same

• US 5,088,499 A
• Filed: 08/20/1990
• Issued: 02/18/1992
• Est. Priority Date: 12/22/1989
• Status: Expired due to Term

• Current Assignee: Lantheus Medical Imaging, Inc. (Lantheus Holdings, Inc.)
• Original Assignee: Evan C. Unger
• Inventors: Unger, Evan C.
• Primary Examiner(s): Howell, Kyle L.
• Assistant Examiner(s): Akers, Scott R.
• Application Number:
  US07/569,828
• Time in Patent Office: 547 Days
• Field of Search: 128/662.02, 128/654, 424/44, 424/9, 424/43, 424/450, 436/829, 264/4.1, 264/4.6
• US Class Current: 424/9.51
• 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

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 of providing an image of an internal region of a patient comprising:

(a) administering to the patient a contrast agent selected from the group consisting of (i) a liposome having encapsulated therein a pH-activated gaseous precursor, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane, (ii) a liposome having encapsulated therein a photo-activated gaseous precursor, and (iii) a liposome having encapsulated therein a temperature-activated gaseous precursor;

(b) activating the gaseous precursor; and

(c) scanning the patient using ultrasonic imaging to obtain enhanced visible images of the region, due to the resulting gas containing liposomes.

2. The method according to claim 1 wherein said liposomes are administered intravascularly and have a mean outer diameter of between about 30 nanometers and about 2 microns.

3. The method according to claim 2 wherein the patient is scanned in the area of the patient'"'"'s heart.

4. The method according to claim 2 wherein the patient is scanned in the area of the patient'"'"'s liver, spleen and kidney.

5. The method according to claim 1 wherein the liposomes are administered other than intravascularly and have a mean outer diameter between about 2 microns and about 10 microns.

6. The method of claim 1 wherein the liposomes are comprised of lipids of either natural or synthetic origin selected from the group consisting of fatty acids, lysolipids, dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, sphingomyelin, lysolipids, cholesterol, cholesterol hemisuccinate, glycosphingolipids, glycolipids, glucolipids, sulphatides, and tocopherol hemisuccinate.

8. A method according to claim 1 wherein the contrast agent is a liposome having encapsulated therein a pH-activated gaseous precursor, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane.

9. A method according to claim 8 wherein said pH-activated gaseous precursor is selected from the group consisting of metal carbonate salts and metal bicarbonate salts.

10. A method according to claim 9 wherein said pH-activated gaseous precursor is sodium bicarbonate.

11. A method according to claim 8 wherein said ionophore in said liposome is selected from the group consisting of carbonylcyanide, p-trifluoromethoxy-phenylhydrazone, carbonylcyanide M-chlorophenylhydrazone, carbonylcyanide phenylhydrazine, tetrachloro-2-trifluoromethyl benzimidazole, 5,6-dichloro-2-trifluoromethyl benzimidazole, Uncoupler 1799, gramicidin, alamethicin, filipin, etruscomycin, nystatin, pimaricin, amphotericin, valinomycin, enniatin, beauvericin, monomycin, nonactin, monactin, dinactin, trinactin, tetranactin, antamanide, nigericin, monensin, salinomycin, narisin, mutalomycin, carriomycin, dianemycin, septamycin, A-204 A, X-206, X-537 A, A-23187 and dicyclohexyl-18-crown-6.

12. A method according to claim 1 wherein the contrast agent comprises a liposome having encapsulated therein a photo-activated gaseous precursor.

13. A method according to claim 12 wherein said photo-activated precursor is a diazonium compound.

14. A method according to claim 1 wherein the contrast agent comprises a liposome having encapsulated therein a temperature-activated gaseous precursor.

15. A method according to claim 14 wherein said temperature-activated gaseous precursor is methyllactate.

7. A method for diagnosing the presence of diseased tissue in a patient comprising:

(a) administering to the patient a contrast agent selected from the group consisting of (i) a liposome having encapsulated therein a pH-activated gaseous precursor, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane, (ii) a liposome having encapsulated therein a photo-activated gaseous precursor, and (iii) a liposome having encapsulated therein a temperature-activated gaseous precursor;

(b) activating the gaseous precursor; and

(c) scanning the patient using ultrasonic imaging to obtain enhanced visible images of any diseased tissue in the patient, due to the resulting gas containing liposomes.

16. A method of providing an image of an internal region of a patient comprising:

(a) administering to the patient a contrast agent selected from the group consisting of (i) a liposome having encapsulated therein a pH-activated gas, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane, (ii) a liposome having encapsulated therein a photo-activated gas, and (iii) a liposome having encapsulated therein a temperature-activated gas; and

(b) scanning the patient using ultrasonic imaging to obtain enhanced visible images of the region, due to the gas containing liposomes.

17. The method according to claim 16 wherein said liposomes are administered intravascularly and have a mean outer diameter of between about 30 nanometers and about 2 microns.

18. The method according to claim 17 wherein the patient is scanned in the area of the patient'"'"'s heart.

19. The method according to claim 17 wherein the patient is scanned in the area of the patient'"'"'s liver, spleen and kidney.

20. The method according to claim 16 wherein the liposomes are administered other than intravascularly and have a mean outer diameter between about 2 microns and about 10 microns.

21. The method of claim 16 wherein the liposomes are comprised of lipids of either natural or synthetic origin selected from the group consisting of fatty acids, lysolipids, dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, sphingomyelin, lysolipids, cholesterol, cholesterol hemisuccinate, glycosphingolipids, glycolipids, glucolipids, sulphatides, and tocopherol hemisuccinate.

22. A method according to claim 16 wherein the contrast agent is a liposome having encapsulated therein a pH-activated gas, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane.

23. A method according to claim 22 wherein said pH-activated gas is gas activated from gaseous precursors selected from the group consisting of metal carbonate salts and metal bicarbonate salts.

24. A method according to claim 23 wherein said pH-activated gas is gas activated from a gaseous precursor which is sodium bicarbonate.

25. A method according to claim 22 wherein said ionophore in said liposome is selected from the group consisting of carbonylcyanide, p-trifluoromethoxy-phenylhydrazone, carbonylcyanide M-chlorophenylhydrazone, carbonylcyanide phenylhydrazine, tetrachloro-2-trifluoromethyl benzimidazole, 5,6-dichloro-2-trifluoromethyl benzimidazole, Uncoupler 1799, gramicidin, alamethicin, filipin, etruscomycin, nystatin, pimaricin, amphotericin, valinomycin, enniatin, beauvericin, monomycin, nonactin, monactin, dinactin, trinactin, tetranactin, antamanide, nigericin, monensin, salinomycin, narisin, mutalomycin, carriomycin, dianemycin, septamycin, A-204 A, X-206, X-537 A, A-23187 and dicyclohexyl-18-crown-6.

26. A method according to claim 16 wherein the contrast agent comprises a liposome having encapsulated therein a photo-activated gas.

27. A method according to claim 26 wherein said photo-activated gas is activated from a gaseous precursor which is a diazonium compound.

28. A method according to claim 16 wherein the contrast agent comprises a liposome having encapsulated therein a temperature-activated gas.

29. A method according to claim 28 wherein said temperature-activated gas is activated from a gaseous precursor which is methyllactate.

30. A method for diagnosing the presence of diseased tissue in a patient comprising:

(a) administering to the patient a contrast agent selected from the group consisting of (i) a liposome having encapsulated therein a pH-activated gas, said liposome having incorporated therein an ionophore, said ionophore being capable of facilitating the transport of hydrogen or hydroxide ions across the liposome membrane, (ii) a liposome having encapsulated therein a photo-activated gas, and (iii) a liposome having encapsulated therein a temperature-activated gas; and

(b) scanning the patient using ultrasonic imaging to obtain enhanced visible images of any diseased tissue in the patient, due to the gas containing liposomes.