A combinatorial in silico and cellular approach to identify a new class of compounds that target VEGFR2 receptor tyrosine kinase activity and angiogenesis

• Published in: British journal of pharmacology 2012-05, Vol.166 (2), p.737-748
• Main Authors: Kankanala, J, Latham, AM, Johnson, AP, Homer‐Vanniasinkam, S, Fishwick, CWG, Ponnambalam, S
• Format: Article
• Language: English
• Subjects: Angiogenesis; Angiogenesis Inhibitors - pharmacology; Biological and medical sciences; Cell adhesion & migration; Cell Proliferation - drug effects; Computer-Aided Design; Drug Design; endothelium; Human Umbilical Vein Endothelial Cells; Humans; Kinases; Medical sciences; Neovascularization, Physiologic - physiology; Pharmacology. Drug treatments; Protein Kinase Inhibitors - pharmacology; Pyrazoles - pharmacology; Receptor, Fibroblast Growth Factor, Type 1 - antagonists & inhibitors; Receptor, Fibroblast Growth Factor, Type 1 - metabolism; Receptor, Fibroblast Growth Factor, Type 3 - antagonists & inhibitors; Receptor, Fibroblast Growth Factor, Type 3 - metabolism; Research Papers; structure‐based design; tyrosine kinase inhibitor; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - pharmacology; Vascular Endothelial Growth Factor Receptor-2 - antagonists & inhibitors; Vascular Endothelial Growth Factor Receptor-2 - metabolism; VEGFR2; Wound Healing - drug effects
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/j.1476-5381.2011.01801.x

BACKGROUND AND PURPOSE Vascular endothelial growth factor receptor 2 (VEGFR2) is an attractive therapeutic target for the treatment of diseases such as cancer. Small‐molecule VEGFR2 inhibitors of a variety of chemical classes are currently under development or in clinical use. In this study, we describe the de novo design of a new generation pyrazole‐based molecule (JK‐P3) that targets VEGFR2 kinase activity and angiogenesis. EXPERIMENTAL APPROACH JK‐P compound series were designed using de novo structure‐based identification methods. Compounds were tested in an in vitro VEGFR2 kinase assay. Using primary endothelial cells, JK‐P compounds were assessed for their ability to inhibit VEGF‐A‐stimulated VEGFR2 activation and intracellular signalling. We tested these compounds in cell migration, proliferation and angiogenesis assays. KEY RESULTS JK‐P3 and JK‐P5 were predicted to bind the VEGFR2 kinase domain with high affinity, and both compounds showed pronounced inhibition of endogenous VEGFR2 kinase activity in primary human endothelial cells. Only JK‐P3 inhibited VEGF‐A‐stimulated VEGFR2 activation and intracellular signalling. Interestingly, JK‐P3 inhibited endothelial monolayer wound closure and angiogenesis but not endothelial cell proliferation. Both compounds inhibited fibroblast growth factor receptor kinase activity in vitro, but not basic fibroblast growth factor‐mediated signalling in endothelial cells. CONCLUSIONS AND IMPLICATIONS This is the first report that describes an anti‐angiogenic inhibitor based on such a pyrazole core. Using a de novo structure‐based identification approach is an attractive method to aid such drug discovery. These results thus provide an important basis for the development of multi‐tyrosine kinase inhibitors for clinical use in the near future.