Target-sensitive immunoliposomes- preparation and characterization

• US 4,957,735 A
• Filed: 02/09/1987
• Issued: 09/18/1990
• Est. Priority Date: 06/12/1984
• Status: Expired due to Fees

• Current Assignee: BioWhittaker, Inc. (Lonza Group AG)
• Original Assignee: The University Of Tennessee Research Corporation (University of Tennessee)
• Inventors: Huang, Leaf
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Saunders, David A.
• Application Number:
  US07/012,321
• Time in Patent Office: 1,317 Days
• Field of Search: 264/4.1, 264/4.3, 264/4.6, 424/85, 424/86, 424/87, 424/85.8, 424/85.91, 424/86, 424/427, 424/450, 436/829, 530/389, 530/390
• US Class Current: 424/178.1
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/586 Liposomes, microcapsules or cells
• 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

Novel target-sensitive immunoliposomes were prepared and characterized. In this invention, target specific binding of antibody-coated liposomes was sufficient to induce bilayer destabilization, resulting in a site-specific release of liposome contents.

Unilamellar liposomes were prepared by using a small quantity of palmitoyl IgG (pIgG) to stabilize the bilayer phase of the unsaturated phosphatidylethanol amine (PE) which by itself does not form stable liposomes. A mouse monoclonal IgG antibody to the glycoprotein D (gD) of Herpes Simplex Virus (HSV) and dioleoyl PE were used in one preferred embodiment.

In another preferred embodiment, potentially cytotoxic antiviral drugs were entrapped in target sensitive (TS) immunoliposomes and delivered to HSV infected cells. Potency was as much as 1000 times superior to the free drug and cytotoxicity was decreased by as much as 3000 fold.

Claims

1. Target sensitive immunoliposomes consisting essentially of phosphatidylethanolamine and a stabilizing amount of fatty acid (C₁₂ to C₂₄) derivatized antibody, said antibody having an affinity for the target cell surface which immunoliposomes release their entrapped contents upon binding to a target cell surface.

2. The target sensitive immunoliposomes of claim 1, wherein the phosphatidylethanolamine is selected from the group consisting of dioleoyl phosphatidylethanolamine and transphosphotidylated phosphtidylethanolamine.

3. The target sensitive immunoliposomes of claim 1, wherein the fatty acid is a palmitic acid derivative.

4. The target sensitive immunoliposomes of claim 1, wherein the antibody is IgG.

5. The target sensitive immunoliposomes of claim 1, wherein the antibody is a monoclonal antibody.

6. The target sensitive immunoliposome of claim 5, wherein the monoclonal antibody recognizes the glycoprotein D of the Herpes Simplex Virus (HSV).

7. The target sensitive immunoliposomes of claim 1, wherein the entrapped contents consist essentially of a pharmaceutically acceptable drug.

8. The target sensitive immunoliposomes of claim 7, wherein the pharmaceutically acceptable drug is selected from the group consisting of cytotoxic and antiviral drugs of the nucleoside analog family.

9. The target sensitive immunoliposomes of claim 8, wherein the cyclotoxic and antiviral drugs are selected from the group consisting of fluorodeoxy uridine, iododeoxyuridine, acyclovir and cytosine arabinoside.

10. The target sensitive immunoliposomes of claim 7, in a form suitable for injection.

11. The target sensitive immunoliposomes of claim 7, in a form suitable for ophthalmic application.

12. The target sensitive immunoliposomes of claim 7, in a form suitable for topical applications.

13. The target sensitive immunoliposomes of claim 1, further consisting essentially of palmitoyl IgG and phosphaticlethanolamine in a molar ratio of not less than 2.5×

10^-4.

14. The target sensitive immunoliposomes of claim 10, wherein the minimal coupling stoichimetry of palmitic acid to antibody is 2.2.

15. A method of delivering therapeutic agents to the surface of cells in need of such treatment comprising the steps of:

(a) forming target sensitive immunoliposomes from phosphatidylethanolamine and a stabilizing amount of fatty acid (C₁₂ to C₂₄) derivatized antibody, said antibody having an affinity for the target cell surface;

(b) entrapping one or more therapeutic agents within the immunoliposomes of step (a); and

(c) administering the immunoliposomes prepared in step (b) to a patient in need of such treatment.

16. The method of claim 15, wherein the phosphatidylethanolamine is selected from the group consisting of dioleoyl phosphatidylethanolamine and transphosphotidylated phosphatidylethanolamine.

17. The method of claim 15, wherein the fatty acid is a palmitic acid derivative.

18. The method of claim 15, wherein the antibodys is IgG.

19. The method of claim 15, wherein the antibody is a monoclonal antibody.

20. The method of claim 19, wherein the monoclonal recognizes the glycoprotein D of the Herpes Simplex Virus (HSV).