Ang-2-VEGF-A CrossMab, a novel bispecific human IgG1 antibody blocking VEGF-A and Ang-2 functions simultaneously, mediates potent antitumor, antiangiogenic, and antimetastatic efficacy

Clin Cancer Res. 2013 Dec 15;19(24):6730-40. doi: 10.1158/1078-0432.CCR-13-0081. Epub 2013 Oct 4.

Abstract

Purpose: VEGF-A blockade has been clinically validated as a treatment for human cancers. Angiopoietin-2 (Ang-2) expression has been shown to function as a key regulator of tumor angiogenesis and metastasis.

Experimental design: We have applied the recently developed CrossMab technology for the generation of a bispecific antibody recognizing VEGF-A with one arm based on bevacizumab (Avastin), and the other arm recognizing Ang-2 based on LC06, an Ang-2 selective human IgG1 antibody. The potency of Ang-2-VEGF CrossMab was evaluated alone and in combination with chemotherapy using orthotopic and subcutaneous xenotransplantations, along with metastasis analysis by quantitative real-time Alu-PCR and ex vivo evaluation of vessels, hypoxia, proliferation, and apoptosis. The mechanism of action was further elucidated using Western blotting and ELISA assays.

Results: Ang-2-VEGF-A CrossMab showed potent tumor growth inhibition in a panel of orthotopic and subcutaneous syngeneic mouse tumors and patient or cell line-derived human tumor xenografts, especially at later stages of tumor development. Ang-2-VEGF-A CrossMab treatment led to a strong inhibition of angiogenesis and an enhanced vessel maturation phenotype. Neoadjuvant combination with chemotherapy resulted in complete tumor regression in primary tumor-bearing Ang-2-VEGF-A CrossMab-treated mice. In contrast to Ang-1 inhibition, anti-Ang-2-VEGF-A treatment did not aggravate the adverse effect of anti-VEGF treatment on physiologic vessels. Moreover, treatment with Ang-2-VEGF-A CrossMab resulted in inhibition of hematogenous spread of tumor cells to other organs and reduced micrometastatic growth in the adjuvant setting.

Conclusion: These data establish Ang-2-VEGF-A CrossMab as a promising antitumor, antiangiogenic, and antimetastatic agent for the treatment of cancer.

MeSH terms

  • Angiopoietin-2 / antagonists & inhibitors
  • Angiopoietin-2 / immunology*
  • Animals
  • Antibodies, Bispecific / administration & dosage*
  • Antibodies, Monoclonal, Humanized / administration & dosage
  • Bevacizumab
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Humans
  • Immunoglobulin G / administration & dosage
  • Immunoglobulin G / immunology
  • Mice
  • Neoplasm Metastasis
  • Neoplasms / drug therapy*
  • Neoplasms / immunology
  • Neovascularization, Pathologic / drug therapy*
  • Neovascularization, Pathologic / immunology
  • Vascular Endothelial Growth Factor A / antagonists & inhibitors
  • Vascular Endothelial Growth Factor A / immunology*

Substances

  • Angiopoietin-2
  • Antibodies, Bispecific
  • Antibodies, Monoclonal, Humanized
  • Immunoglobulin G
  • Vascular Endothelial Growth Factor A
  • Bevacizumab