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Details for Patent: 5,305,757

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Details for Patent: 5,305,757

Title: Gas filled liposomes and their use as ultrasonic contrast agents
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.
Inventor(s): Unger; Evan C. (Tucson, AZ), Wu; Guanli (Tucson, AZ)
Assignee:
Filing Date:Feb 12, 1993
Application Number:08/017,683
Claims:1. A method for preparing contrast agents for ultrasonic imaging comprising the following steps:

(i) placing liposomes under negative pressure;

(ii) incubating said liposomes under said negative pressure for a time sufficient to remove substantially all liquid from said liposomes; and

(iii) instilling gas into said liposomes until ambient pressures are achieved.

2. A method of claim 1 further comprising allowing said liposomes to cool prior to and during step (i) to a temperature between about -10.degree. C. and about -20.degree. C., allowing said liposomes to warm during step (ii) to a temperature between about 10.degree. C. and about 20.degree. C., and allowing said liposomes to warm during step (iii) to ambient temperatures.

3. A method of claim 1 wherein said negative pressure is between about 700 mm Hg and about 760 mm Hg and is applied for about 24 to about 72 hours.

4. A method of claim 1 where said gas is instilled into said liposomes over a period of about 4 to about 8 hours.

5. A method of claim 1 where said gas is selected from the group consisting of air, nitrogen, carbon dioxide, oxygen, argon, xenon, neon, and helium.

6. A method of claim 5 where said gas is nitrogen.

7. A method of claim 1 further comprising, after step (iii), extruding said liposomes through at least one filter of a selected pore size.

8. A contrast agent produced by the method of claim 1.

9. A method for preparing contrast agents for ultrasonic imaging comprising the following steps:

(i) allowing liposomes to cool to a temperature between about -10.degree. C. and about -20.degree. C.;

(ii) placing said liposomes under a negative pressure of between about 700 mm Hg to about 760 mm Hg;

(iii) incubating said liposomes under said negative pressure for about 24 to about 72 hours to remove substantially all liquid from said liposomes; while allowing said liposomes to warm to a temperature between about 10.degree. C. and about 20.degree. C.; and

(iv) instilling gas into said liposomes over a period of about 4 to about 8 hours until ambient pressures are achieved, while allowing said liposomes to warm to ambient temperature.

10. A method of claim 9 where said gas is selected from the group consisting of air, nitrogen, carbon dioxide, oxygen, argon, xenon, neon, and helium.

11. A method of claim 10 where said gas is nitrogen.

12. A method of claim 9 further comprising, after step (iv), extruding said liposomes through at least one filter of a selected pore size.

13. A contrast agent produced by the method of claim 9.

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

(a) administering to the patient a contrast agent comprising gas filled liposomes prepared by a vacuum drying gas instillation method; and

(b) scanning the patient using ultrasonic imaging to obtain visible images of the region.

15. A method according to claim 14 wherein the patient is scanned in the area of the patient's left heart.

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

(a) administering to the patient a contrast agent comprising gas filled liposomes substantially devoid of liquid in the interior thereof; and

(b) scanning the patient using ultrasonic imaging to obtain visible images of the region.

17. A method according to claim 16 wherein the patient is scanned in the area of the patient's left heart.

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

(i) administering to the patient a contrast agent comprising gas filled liposomes prepared by a vacuum drying gas instillation method; and

(ii) scanning the patient using ultrasonic imaging to obtain visible images of any diseased tissue in the patient.

19. A method according to claim 18 wherein the patient is scanned in the area of the patient's left heart.

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

(i) administering to the patient a contrast agent comprising gas filled liposomes substantially devoid of liquid in the interior thereof; and

(ii) scanning the patient using ultrasonic imaging to obtain visible images of any diseased tissue in the patient.

21. A method according to claim 20 wherein the patient is scanned in the area of the patient's left heart.

22. A method for preparing gas filled liposomes comprising the following steps:

(i) placing liposomes under negative pressure;

(ii) incubating said liposomes under said negative pressure for a time sufficient to remove substantially all liquid from said liposomes; and

(iii) instilling gas into said liposomes until ambient pressures are achieved.

23. A gas filled liposome produced by the method of claim 22.

24. A method for preparing gas filled liposomes comprising the following steps:

(i) allowing liposomes to cool to a temperature between about -10.degree. C. and about -20.degree. C.;

(ii) placing said liposomes under a negative pressure of between about 700 mm Hg to about 760 mm Hg;

(iii) incubating said liposomes under said negative pressure for about 24 to about 72 hours to remove substantially all liquid from said liposomes; while allowing said liposomes to warm to a temperature between about 10.degree. C. and about 20.degree. C.; and

(iv) instilling gas into said liposomes over a period of about 4 to about 8 hours until ambient pressures are achieved, while allowing said liposomes to warm to ambient temperature.

25. A gas filled liposome produced by the method of claim 24.
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