Novel 4-Anilinoquinazolines with C-7 Basic Side Chains: Design and Structure Activity Relationship of a Series of Potent, Orally Active, VEGF Receptor Tyrosine Kinase Inhibitors
journal contributionposted on 19.02.2002, 00:00 by Laurent F. Hennequin, Elaine S. E. Stokes, Andrew P. Thomas, Craig Johnstone, Patrick A. Plé, Donald J. Ogilvie, Michael Dukes, Stephen R. Wedge, Jane Kendrew, Jon O. Curwen
We have previously shown that 4-anilinoquinazolines can be potent inhibitors of vascular endothelial growth factor (VEGF) receptor (Flt-1 and KDR) tyrosine kinase activity. A novel subseries of 4-anilinoquinazolines that possess basic side chains at the C-7 position of the quinazoline nucleus have been synthesized. This subseries contains potent, nanomolar inhibitors of KDR (median IC50 0.02 μM, range 0.001−0.04 μM), which are comparatively less potent vs Flt-1 tyrosine kinase (median IC50 0.55 μM, range 0.02−1.6 μM). The compounds also retain some inhibitory activity against the tyrosine kinase associated to the endothelial growth factor receptor (EGFR) (median IC50 0.2 μM, range 0.075−0.8 μM) but demonstrate selectivity vs that associated to the FGF receptor 1 (median IC50 2.5 μM, range 0.9−19 μM). This selectivity profile is also evident in a growth factor-stimulated human endothelial cell (HUVEC) proliferation assay (i.e., inhibition of VEGF > EGF > FGF), with inhibition of VEGF-induced proliferation being achieved at nanomolar concentrations (median IC50 0.06 μM). Further examination of compound 2 (ZD6474) in recombinant enzyme assays revealed excellent selectivity for the inhibition of KDR tyrosine kinase (IC50 0.04 μM) vs the kinase activity of erbB2, MEK, CDK-2, Tie-2, IGFR-1R, PDK, PDGFRβ, and AKT (IC50 range: 1.1 to >100 μM). Anilinoquinazolines possessing basic C-7 side chains exhibited markedly improved aqueous solubility over previously described anilinoquinazolines possessing neutral C-7 side chains (up to 500-fold improvement at pH 7.4). In addition, aqueous solubility of the neutral fraction present at pH 7.4 of the basic subseries of anilinoquinazoline proved to be higher than that of the neutral analogue 1 (ZD4190). Oral administration of representative compounds to mice (50 mg/kg) produced plasma levels between 0.2 and 3 μM at 24 h after dosing. Our development candidate 2 demonstrated a very attractive in vitro profile combined with excellent solubility (330 μM at pH 7.4) and good oral bioavailability in rat and dog (>80 and >50%, respectively). This compound demonstrated highly significant, dose-dependent, antitumor activity in athymic mice. Once daily oral administration of 100 mg/kg of compound 2 for 21 days inhibited the growth of established Calu-6 lung carcinoma xenografts by 79% (P < 0.001, Mann Whitney rank sum test), and substantial inhibition (36%, P < 0.02) was evident with 12.5 mg/kg/day.
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IC 50 2.5 μ Mcompound 2IC 50 0.02 μ MIC 50 rangeStructure Activity RelationshipsolubilityIC 50 0.04 μ MAKT3 μ Mtyrosine kinaseanilinoquinazolineselectivityIGFRgrowth factor receptorIC 50 0.2 μ MZDPDGFRIC 50 0.55 μ Mdevelopment candidate 2pHKDR tyrosine kinaseFGF receptor 1PDKmgMEKCDKHUVECEGFRIC 50 0.06 μ Mvssubserietyrosine kinase activityVEGF Receptor Tyrosine Kinase Inhibitorsinhibition