Apparatus for preparing gas filled liposomes for use as ultrasonic contrast agents

• US 5,348,016 A
• Filed: 07/07/1993
• Issued: 09/20/1994
• 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, Guanli Wu
• Inventors: Wu, Guanli, Unger, Evan C.
• Primary Examiner(s): JAWORSKI, FRANCIS J
• Application Number:
  US08/088,268
• Time in Patent Office: 440 Days
• Field of Search: 436/829, 128/662.02, 264/4.1, 264/4.6, 428/402.2, 424/450
• US Class Current: 600/448
• 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. An apparatus for preparing contrast agents for ultrasonic imaging by drying liposomes and instilling gas therein comprising:

(i) a vessel for containing liposomes;

(ii) means for sequentially applying a negative pressure to the vessel for a first pre-determined period of time to draw liquid from liposomes contained in the vessel and then subsequently applying a gas to the vessel to gradually eliminate the negative pressure therein over a second pre-determined period of time, the means for sequentially applying the negative pressure and then the gas to gradually eliminate the negative pressure including (a) a vacuum source, (b) a source of pressurized gas, and (c) a conduit having means for selectively placing the vacuum source and the pressurized gas in flow communication with the vessel.

2. An apparatus of claim 1, wherein the vacuum source is a vacuum pump.

3. An apparatus according to claim 1 further comprising means for cooling the liposomes contained in the vessel.

4. An apparatus according to claim 3 wherein the cooling means has means for cooling liposomes contained in the vessel to between about -10°

C. and about -20°

C.

5. An apparatus according to claim 4 wherein the cooling means comprises an ice bath.

6. An apparatus according to claim 1 further comprising means for collecting the liquid drawn from the liposomes.

7. An apparatus according to claim 6 wherein the collecting means is a trap.

8. An apparatus according to claim 7 further comprising means for cooling the trap.

9. An apparatus according to claim 7 wherein the trap comprises first and second members adapted to direct fluid flow therethrough, the members in flow communication with each other and with the conduit, the second member being helically arranged around the first member, the trap comprising a cooling means wherein the cooling means comprises an ice bath enclosing at least a portion of the first and second members.

10. An apparatus for preparing contrast agents for ultrasonic imaging by drying liposomes and instilling gas therein comprising:

(i) a vessel for containing liposomes;

(ii) means for cooling liposomes contained in the vessel to a temperature at least as low as approximately 0°

C.;

(iii) means for applying a negative pressure to the vessel at least as low as approximately 700 mm Hg for a period of time at least as long as approximately 24 hours to draw liquid from the liposomes contained in the vessel;

(iv) a conduit connecting the negative pressurizing means to the vessel, the conduit directing the flow of the liquid;

(v) means for collecting the liquid flowing in the conduit; and

(vi) means for isolating the negative pressurizing means from the vessel and means for thereafter gradually introducing a gas into liposomes contained in the vessel so as to gradually eliminate the negative pressure therein over a period of at least 4 hours.

11. An apparatus according to claim 10, wherein the means for thereafter gradually introducing a gas into liposomes contained in the vessel comprises a source of pressurized gas and a valve, and wherein the conduit further connects the source of pressurized gas to the vessel, said valve being disposed in said conduit.

12. An apparatus for drying liposomes and instilling gas therein comprising:

(i) a vessel for containing liposomes;

(ii) means for sequentially applying a negative pressure to the vessel for a first pre-determined period of time to draw liquid from liposomes contained in the vessel and then subsequently applying a gas to the vessel to gradually eliminate the negative pressure therein over a second pre-determined period of time, the means for sequentially applying the negative pressure and then the gas to gradually eliminate the negative pressure including (a) a vacuum source, (b) a source of pressurized gas, and (c) a conduit having means for selectively placing the vacuum source and the pressurized gas in flow communication with the vessel.

13. An apparatus for drying liposomes and instilling gas therein comprising:

(i) a vessel for containing liposomes;

(ii) means for cooling liposomes contained in the vessel to a temperature at least as low as approximately 0°

C.;

(iii) means for applying a negative pressure to the vessel of at least approximately 700 mm Hg for a period of at least as long as approximately 24 hours to draw liquid from liposomes contained in the vessel;

(iv) a conduit connecting the negative pressurizing means to the vessel, the conduit directing the flow of the liquid;

(v) means for collecting the liquid flowing in the conduit; and

(vi) means for isolating the negative pressurizing means from said vessel and for thereafter gradually introducing a gas into liposomes contained in said vessel so as to gradually eliminate the negative pressure therein over a period of at least 4 hours.

14. An apparatus according to claim 13, wherein the means for isolating said negative pressurizing means and for thereafter gradually introducing a gas into liposomes contained in the vessel comprises a source of pressurized gas and a valve, and wherein the conduit further connects the source of pressurized gas to the vessel, said valve being disposed in said conduit.