Delivery of biologically active substance to target sites in the body of patients

• US 6,258,378 B1
• Filed: 02/03/1999
• Issued: 07/10/2001
• Est. Priority Date: 02/09/1998
• Status: Expired due to Term

• Current Assignee: Bracco Suisse SA
• Original Assignee: Bracco Research SA
• Inventors: Hiver, Agnés, Yan, Feng, Schneider, Michel
• Primary Examiner(s): Kishore, Gollamudi S.
• Application Number:
  US09/243,787
• Time in Patent Office: 888 Days
• Field of Search: 424/450, 424/1.21, 424/9.321, 424/9.51, 424/812, 436/829
• US Class Current: 424/450
• 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 9/0009 involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis microelectromechanical systems A61K9/0097
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: Y10S 424/812 Liposome comprising an antibody, antibody fragment, antigen, or other specific or nonspecific immunoeffector
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

The invention concerns a method for administering bioactive substances to patients at selected sites in the body and remotely promoting delivery of said media to selected organs or tissues in the body. The method comprises providing an administrable formulation comprising, dispersed in an aqueous carrier liquid, liposomes filled with bioactive substances and gas-filled microspheres, injecting said formulation into the circulation of a patient so that it is directed to a site of interest, and applying ultrasound pulses to said site so as to make the gas-filled microbodies explode and the gas confined therein to expand in the carrier liquid, the energy of expansion of said confined gas causing the liposome vesicles to open and release the trapped substances at said site. Also disclosed are formulations for delivery of biologically active substances to selected target sites in the organism, the formulations comprising an aqueous suspension of gas-filled microvesicles and liposomes filled with active susbstances such as drugs or diagnostic agents. The formulations are available in a kit form in which the kit comprises sterile precursor components.

Claims

1. A method of delivering a drug or diagnostic agent to a selected site in the body of a patient comprising the steps of:

(i) injecting into the circulation of said patient an administrable formulation consisting essentially of, as a suspension in a carrier liquid, microballoons bounded by a solid polymeric membrane carrying an entrapped physiologically acceptable gas and liposome vesicles filled with a liquid containing said drug or diagnostic agent;

(ii) allowing said formulation to reach the selected site through the circulation of said patient, and

(iii) ultrasonically irradiating said site so as to cause the microballoons to explode and the entrapped gas to expand in the carrier liquid, the energy of expansion of said gas causing the liposome vesicles to open and release the drug or diagnostic agent at said site.

3. The method of claim 1 or 2, wherein the microballoons or microbubbles are exploded by irradiation with ultrasonic pulses having frequencies of about 0.3 to 3 MHz.

4. The method of claim 3, wherein the ultrasonic pulses are provided by a standard or a modified echography probe adapted to simultaneously monitor the reflected echo signal and thereby provide an image of the irradiated site.

9. The method of claim 1, wherein the membrane of the microballoons is made from a natural or a synthetic polymer.

10. The formulation of claim 1 or 2, wherein the entrapped physiologically acceptable gas is selected from the group consisting of SF₆, CF₄, C₂F₆, C₃F₆, C₃F₈, C₄F₆, C₄F₈, C₄F₁₀, C₅F₁₀, C₅F₁₂, C₆F₁₄, air, oxygen, nitrogen, carbon dioxide, noble gases, and mixtures thereof.

12. The method of claim 1 or 2, wherein the liposome vesicles and the air or gas-filled microballoons or microbubbles have affinity for each other.

13. The method of claim 12, wherein the liposome vesicles and the microspheres are each provided with the respective components of a conjugate pair.

14. The method of claim 13, wherein an antigen is present in the liposome membrane and an antibody in the gas-filled component, or vice-versa, so that antigen-antibody conjugation will cause the liposome vesicles and the microballoons or microbubbles to be brought together.

15. The method of claim 12, wherein both the liposome vesicles and the microballoons or microbubbles are provided with a donor coupler element and the formulation further includes a multisite acceptor element, whereby donor and acceptor will become conjugated and the liposome vesicles and the microballoons or microbubbles are brought together.

2. A method of delivering a drug or diagnostic agent to a selected site in the body of a patient comprising the steps of:

(i) injecting into the circulation of said patient an administrable formulation consisting essentially of, as a suspension in a carrier liquid, microbubbles bounded by an evanescent gas/liquid interfacial closed surface and carrying an entrapped physiologically acceptable gas and liposome vesicles filled with a liquid containing said drug or diagnostic agent;

(ii) allowing said formulation to reach the selected site through the circulation of said patient, and

(iii) ultrasonically irradiating said site so as to cause the microbubbles to explode and the entrapped gas to expand in the carrier liquid, the energy of expansion of said gas causing the liposome vesicles to open and release the drug or diagnostic agent at said site.

5. The method of claim 2, wherein the carrier liquid comprises amphipatic compounds to stabilize the gas-containing microbubbles against premature collapse.

6. The method of claim 5, wherein the amphipatic compounds are phospholipids.

7. The method of claim 6, wherein the phospholipids are saturated.

8. The method of claim 6, wherein the microbubbles are stabilized by a monolayer of phospholipids at the gas/liquid interface.

11. The method of claim 6, wherein the phospholipids are neutral phospholipids selected from the group consisting of hydrogenated phosphatidyl, choline (HSPC), dipalmitoyl-, distearoyl- and diarachidoyl phosphatidylcholine (DPPC, DSPC, DAPC), negatively charged phospholipids selected from the group consisting of dipalmitoyl and distearoyl phosphatidic acid (DPPA), DSPA), dipalmitoyl and distearoyl phosphatidylserine (DPPS, DSPS), dipalmitoyl and distearoyl phosphatidylglycerol (DPPG, DSPG);

reactive phospholipids selected from the group consisting of phosphatidyl ethanolamine derivatives coupled to a polyethyleneglycol, a biotinyl, a glutaryl, a caproyl or a succinyl amine.