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[Cancer Research 62, 4645-4655, August 15, 2002]
© 2002 American Association for Cancer Research

Experimental Therapeutics

ZD6474 Inhibits Vascular Endothelial Growth Factor Signaling, Angiogenesis, and Tumor Growth following Oral Administration

Stephen R. Wedge1, Donald J. Ogilvie, Michael Dukes, Jane Kendrew, Rosemary Chester, Janet A. Jackson, Sarah J. Boffey, Paula J. Valentine, Jon O. Curwen, Helen L. Musgrove, George A. Graham, Gareth D. Hughes, Andrew P. Thomas, Elaine S. E. Stokes, Brenda Curry, Graham H. P. Richmond, Peter F. Wadsworth, Alison L. Bigley and Laurent F. Hennequin

Departments of Cancer and Infection Research [S. R. W., D. J. O., M. D., J. K., R. C., J. A. J., S. J. B., P. J. V., J. O. C., H. L. M., G. A. G., G. D. H., A. P. T., E. S. E. S., B. C.] and Safety Assessment [G. H. P. R., P. F. W., A. L. B.], AstraZeneca, Cheshire SK10 4TG, United Kingdom, and AstraZeneca Pharma, Centre de Recherches, 51689 Reims Cedex 2, France [L. F. H.]

ZD6474 [N-(4-bromo-2-fluorophenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine]is a potent, p.o. active, low molecular weight inhibitor of kinase insert domain-containing receptor [KDR/vascular endothelial growth factor receptor (VEGFR) 2] tyrosine kinase activity (IC50 = 40 nM). This compound has some additional activity versus the tyrosine kinase activity of fms-like tyrosine kinase 4 (VEGFR3;IC50 = 110 nM) and epidermal growth factor receptor (EGFR/HER1; IC50 = 500 nM) and yet demonstrates selectivity against a range of other tyrosine and serine-threonine kinases. The activity of ZD6474 versus KDR tyrosine kinase translates into potent inhibition of vascular endothelial growth factor-A (VEGF)-stimulated endothelial cell (human umbilical vein endothelial cell) proliferation in vitro (IC50 = 60 nM). Selective inhibition of VEGF signaling has been demonstrated in vivo in a growth factor-induced hypotension model in anesthetized rat: administration of ZD6474 (2.5 mg/kg, i.v.) reversed a hypotensive change induced by VEGF (by 63%) but did not significantly affect that induced by basic fibroblast growth factor. Once-daily oral administration of ZD6474 to growing rats for 14 days produced a dose-dependent increase in the femoro-tibial epiphyseal growth plate zone of hypertrophy, which is consistent with inhibition of VEGF signaling and angiogenesis in vivo. Administration of 50 mg/kg/day ZD6474 (once-daily, p.o.) to athymic mice with intradermally implanted A549 tumor cells also inhibited tumor-induced neovascularization significantly (63% inhibition after 5 days; P < 0.001). Oral administration of ZD6474 to athymic mice bearing established (0.15–0.47 cm3), histologically distinct (lung, prostate, breast, ovarian, colon, or vulval) human tumor xenografts or after implantation of aggressive syngeneic rodent tumors (lung, melanoma) in immunocompetent mice, produced a dose-dependent inhibition of tumor growth in all cases. Statistically significant antitumor activity was evident in each model with at least 25 mg/kg ZD6474 once daily (P < 0.05, one-tailed t test). Histological analysis of Calu-6 tumors treated with 50 mg/kg/day ZD6474 for 24 days showed a significant reduction (>70%) in CD31 (endothelial cell) staining in nonnecrotic regions. ZD6474 also restrained growth of much larger (0.9 cm3 volume) Calu-6 lung tumor xenografts and induced profound regression in established PC-3 prostate tumors of 1.4 cm3 volume. ZD6474 is currently in Phase I clinical development as a once-daily oral therapy in patients with advanced cancer.




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