In vitro study | PHA-665752 significantly inhibited c-Met kinase activity with a Ki of 4 nM and an IC50 of 9 nM and a 50-fold higher selectivity for c-Met compared to various tyrosine and serine-threonine kinases. PHA-665752 effective inhibition of HGF stimulated c-Met autophosphorylation, IC50 25-50 nM. PHA-665752 also significantly inhibited HGF-and c-Met-dependent functions such as cell motility and cell proliferation with an IC50 of 40-50 nM and 18-42 nM, respectively. In addition, PHA-665752 acts on a variety of tumor cell lines, effectively inhibiting phosphorylation of HGF-stimulated or constitutive c-Met of downstream regulators such as Gab-1, ERK, Akt, STAT3, PLC-γ, and FAK. PHA-665752 inhibited cell growth in TPR-MET-transformed BaF3 cells in a dose-dependent manner, and 0.2 μm PHA-665752 also inhibited constitutive cell viability and migration with an inhibition rate of 92.5%. Inhibition of 0.2 μm PHA665752 by c-Met also induced apoptosis in 33.1% cells and increased G1 phase cells from 42.4% to 77.0%. PHA-665752 significantly inhibited c-Met kinase activity with K 1 at 4 nM and IC50 at 9 nM, and was 50-fold more selective for c-Met than various tyrosine and serine-threonine kinases. PHA-665752 effective inhibition of HGF stimulated c-Met autophosphorylation, IC50 25-50 nM. PHA-665752 also significantly inhibited HGF-and c-Met-dependent functions such as cell motility and cell proliferation with an IC50 of 40-50 nM and 18-42 nM, respectively. In addition, PHA-665752 acts on a variety of tumor cell lines, effectively inhibiting phosphorylation of HGF-stimulated or constitutive c-Met of downstream regulators such as Gab-1, ERK, Akt, STAT3, PLC-γ, and FAK. PHA-665752 inhibited cell growth in TPR-MET-transformed BaF3 cells in a dose-dependent manner, and 0.2 μm PHA-665752 also inhibited constitutive cell viability and migration with an inhibition rate of 92.5%. 0.2 M PHA665752 inhibited c-Met, also induced 33.1% cell apoptosis, and the G1 phase cells from 42.4% Increased to 77.0%. |
In vivo study | Consistent with c-Met inhibition of phosphorylation and signal transduction in vivo, PHA-665752 treatment of S114 xenografts inhibited tumor growth in a dose-dependent manner, with doses of 7.5, 15, and 30 mg/kg daily, tumor growth inhibition was 20%, 39% and 68%, respectively. Treatment of the mouse xenograft tumor model with PHA665752 significantly reduced tumor growth by 99%,75%, and 59% in NCI-H69, NCI-H441, and A549, respectively. PHA665752 also significantly inhibited angiogenesis by more than 85%, decreased the production of endothelial growth factor, and increased the production of angiogenesis inhibitor platelet -1. consistent with c-Met inhibition of phosphorylation and signal transduction in vivo, PHA-665752 treatment of S114 xenografts inhibited tumor growth in a dose-dependent manner, with 7.5, 15, and 30 mg/kg dose treatment, tumor growth inhibition was 20%, 39% and 68%, respectively. Treatment of the mouse xenograft tumor model with PHA665752 significantly reduced tumor growth by 99%,75%, and 59% in NCI-H69, NCI-H441, and A549, respectively. PHA665752 also significantly inhibited angiogenesis by more than 85%, decreased the production of endothelial growth factor, and increased the production of angiogenesis inhibitor platelet -1. |