Anti-vascular endothelial growth factor receptor 2 (VEGFR2) antibody and methods of use thereof for detecting VEGFR2 and for inhibiting tumor growth, tumor angiogenesis and/or inducing cancer cell cytotoxicity

Patent metadata

Field	Value
Publication number	US-10196447-B2
Application number	US-201615563950-A
Country	US
Kind code	B2
Filing date	Apr 12, 2016
Priority date	Apr 14, 2015
Publication date	Feb 5, 2019
Grant date	Feb 5, 2019

Abstract

An isolated antibody or an antigen-binding fragment thereof having a specific binding affinity to an epitope located within the domain 1 or domain 3 of human vascular endothelial growth factor receptor 2 (VEGFR2; SEQ ID NO: 74) is disclosed. The epitope within the domain 3 of the VEGFR2 is located between amino acid residues 250 and 270 of SEQ ID NO: 74. Use of the antibody or antigen-binding fragment thereof in the manufacture of a medicament for inhibiting tumor growth, tumor angiogenesis, and/or inducing cancer cell cytotoxicity in a subject in need thereof is also disclosed. Also disclosed is a method of detecting the presence of VEGFR2 in a tumor vascular endothelial cell or a cancer cell in a biological sample.

First claim

What is claimed is:

1. An isolated antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region (V H ) and a light chain variable region (V L ), the V H comprising V H CDR1, V H CDR2, and V H CDR3, and the V L comprising V L CDR1, V L CDR2, and V L CDR3,
wherein:
the V H CDR1, V H CDR2, V H CDR3 comprise the amino acid sequence of SEQ NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, respectively; and the V L CDR1, V L CDR2, and V L CDR3 comprise the amino acid sequence of SEQ ID NO: 9, Asp Ala Ser, and SEQ ID NO: 10 or 73, respectively.

2. The antibody or antigen-binding fragment thereof of claim 1 , wherein:
the V L CDR3 comprises the amino acid sequence of SEQ ID NO: 73.

3. The antibody or antigen-binding fragment thereof of claim 1 , wherein
the V H comprises the amino acid sequence of SEQ ID NO: 76; and
the V L comprises the amino acid sequence of SEQ ID NO: 77 or 78.

4. The antibody or antigen-binding fragment thereof of claim 1 , which is a single-chain variable fragment, a Fab fragment or a Fv fragment.

5. The antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody is a fully human antibody.

6. A composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 5 and a pharmaceutically acceptable vehicle or carrier.

7. The composition of claim 6 , further comprising a therapeutically effective amount of a chemotherapeutic agent.

8. The antibody or antigen-binding fragment thereof of claim 3 , wherein the antibody is a fully human antibody.

9. The antibody or antigen-binding fragment thereof of claim 4 , wherein the antibody is a fully human antibody.

10. The antibody or antigen-binding fragment thereof of claim 2 , wherein the antibody is a fully human antibody.

11. An isolated antibody or an antigen-binding fragment thereof, or a single-chain variable fragment, which comprises:
(a) a heavy chain variable region (V H ) comprising the amino acid sequence of SEQ ID NO: 76; and
(b) a light chain variable region (V L ) comprising the amino acid sequence of SEQ ID NO: 77 or 78.

12. A method of detecting the presence of VEGFR2 on tumor vascular endothelial cells or cancer cells in a biological sample, comprising:
(i) admixing the antibody or antigen-binding fragment thereof of claim 1 with the biological sample;
(ii) allowing the antibody or antigen-binding fragment thereof and the VEGFR2 on the tumor vascular endothelial cells or cancer cells in the biological sample to interact and form a complex; and
(iii) detecting the presence of the VEGFR2 on the tumor vascular endothelial cells or cancer cells in the complex.

13. A method of detecting the presence of VEGFR2 on tumor vascular endothelial cells or cancer cells in a biological sample, comprising:
(i) admixing the antibody or antigen-binding fragment thereof of claim 2 with the biological sample;
(ii) allowing the antibody or antigen-binding fragment thereof and the VEGFR2 on the tumor vascular endothelia cells or cancer cells in the biological sample to interact and from a complex; and
(iii) detecting the presence of the VEGFR2 on the tumor vascular endothelial cells or cancer cells in the complex.

14. A method for inhibiting tumor growth, tumor angiogenesis, and/or inducing cytotoxicity in cancer cells, comprising:
administering to a subject in need thereof a composition comprising a therapeutically effective amount of the isolated antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier, wherein the tumor and/or cancer cell expresses VEGFR2.

15. A method for inhibiting tumor growth, tumor angiogenesis, and/or inducing cancer cell cytotoxicity in a subject in need thereof, comprising:
administering to the subject in need thereof a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 2 , wherein the antibody is a fully human antibody.

16. The method of claim 15 , wherein the tumor or cancer is at least one selected from the group consisting of pancreatic, breast, lung, leukemia, prostate and ovary cancer.

17. A method for inhibiting tumor growth, tumor angiogenesis, and/or inducing cancer cell cytotoxicity in a subject in need thereof, comprising:
administering to the subject in need thereof a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 5 .

18. The method of claim 17 , wherein the tumor or cancer is at least one selected from the group consisting of pancreatic, breast, lung, leukemia, prostate and ovary cancer.

19. A method for inhibiting tumor growth, tumor angiogenesis, and/or inducing cancer cell cytotoxicity in a subject in need thereof, comprising:
administering to the subject in need thereof a therapeutically effective amount of the antibody or antigen-binding fragment thereof of claim 8 .

20. The method of claim 19 , wherein the tumor or cancer is at least one selected from the group consisting of pancreatic, breast, lung, leukemia, prostate and ovary cancer.

Assignees

• Academia Sinica

Inventors

• Wu Han-Chung
• Lu Ruei-Min
• Chiu Chiung-Yi
• Liu I-Ju
• Chang Yu-Ling

Classifications

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K2317/73

  Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation · CPC title

• A61P35/02

  specific for leukemia · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title