IN VIVO SCREENING ASSAYS

• US 20110044907A1
• Filed: 08/20/2010
• Published: 02/24/2011
• Est. Priority Date: 08/21/2009
• Status: Abandoned Application

• Current Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Original Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Inventors: Smith, Victoria, Marshall, Derek
• Application Number:
  US12/860,693
• Publication Number:
  US 20110044907A1
• Time in Patent Office: Days
• US Class Current: 424/9.2
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: A61K 49/0047 Green fluorescent protein [GFP]

Abstract

Disclosed herein are methods for identifying inhibitors of LOXL2 activity using in vivo assays. The assays include, for example, CCL₄-induced liver fibrosis, bleomycin-induced lung fibrosis, collagen-induced arthritis, tumor growth following surgical orthotopic implantation of cultured tumor cells, metastasis following intracardiac injection of cultured tumor cells, tumor xenograft models, and vasculogenesis and/or angiogenesis following implantation of exogenous basement membrane.

Claims

1. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise one or more areas of fibrosis, and wherein a test molecule that ameliorates symptoms of fibrosis is identified as an inhibitor of LOXL2 activity.

2. The method of claim 1, wherein the fibrosis is liver fibrosis.

3. The method of claim 2, wherein the fibrosis is induced by CCl₄ treatment.

4. The method of claim 1, wherein the fibrosis is lung fibrosis.

5. The method of claim 4, wherein the fibrosis is induced by treatment with bleomycin.

18. The method of any of claim 1, 6, 8, 11, 13, or 15, wherein the test molecule is a polypeptide.

19. The method of claim 18, wherein the polypeptide is an antibody.

20. The method of claim 19, wherein the antibody is an anti-LOXL2 antibody.

21. The method of any of claim 1, 6, 8, 11, 13 or 15, wherein the test molecule is a nucleic acid.

22. The method of claim 21, wherein the nucleic acid is a siRNA.

6. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise one or more areas of arthritis, and wherein a test molecule that ameliorates symptoms of arthritis is identified as an inhibitor of LOXL2 activity.

7. The method of claim 6, wherein the arthritis is induced by injection of collagen.

8. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise one or more experimental tumors generated by surgical orthotopic implantation of tumor cells, and wherein a test molecule that reduces tumor volume is identified as an inhibitor of LOXL2 activity.

9. The method of claim 8, wherein the tumor cells are MDA-MB435 cells.

10. The method of claim 8, wherein the experimental tumor is a lung tumor.

11. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise experimental metastases generated by intravascular injection of tumor cells, and wherein a test molecule that reduces the degree of metastasis is identified as an inhibitor of LOXL2 activity.

12. The method of claim 11, wherein the tumor cells are MDA-MB231 cells.

13. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise an exogenous basement membrane, and wherein a test molecule that reduces vasculogenesis of the exogenous basement membrane is identified as an inhibitor of LOXL2 activity.

14. The method of claim 13, wherein the exogenous basement membrane comprises Matrigel.

15. A method for identifying an inhibitor of LOXL2 activity, the method comprising treating animals with a test molecule, wherein the animals comprise one or more areas of desmoplasia, and wherein a test molecule that ameliorates symptoms of desmoplasia is identified as an inhibitor of LOXL2 activity.

16. The method of claim 15, wherein amelioration of symptoms of desmoplasia is evidenced by reduction in collagen crosslinking.

17. The method of claim 15, wherein amelioration of symptoms of desmoplasia is evidenced by reduction in expression of alpha-smooth muscle actin.