SU11274 (Met Kinase Inhibitor)(IC50=10 nM) is a specific Met inhibitor. It shows no significant effects on PGDFRβ, EGFR or Tie2.

MET:0.01 μM

SU11274抑制PI3K通路关键调节器的磷酸化, 包括AKT, FKHR,或GSK3β。SU11274抑制HGF依赖的Met磷酸化,及HGF依赖的细胞增殖和活动,IC50为1-1.5 μM。在TPR-MET转化的BaF3细胞中,白细胞介素- 3存在时,用SU11274处理细胞,以剂量依赖的方式抑制细胞生长。SU11274（IC50<3 μM）则不会抑制其他致癌酪氨酸激酶转化的BaF3细胞生长,包括BCR-ABL, TEL-JAK2, TEL-ABL,和TEL-PDGFβR。在BaF3.TPR-MET细胞中,经SU11274处理,也显著抑制细胞迁移,按1 μM 和5 μM处理,抑制分别为44.8% 和 80%。在具有功能性Met受体的H69和H345细胞中,SU11274抑制 HGF诱导的细胞生长,IC50分别为3.4 μM 和 6.5 μM。表达c-Met的非小细胞肺癌(NSCLC)细胞中,SU1127抑制细胞活力,IC50为0.8-4.4 μM,同时去除肝脏生长因子诱导的c-Met 及其下游信号磷酸化。

In vitro Met kinase assay: A chimeric protein is constructed containing the cytoplasmic domain of human c-Met fused to Glutathione S-transferase (GST) and expressed in SF9 cells. The c-Met kinase GST-fusion protein is used for an ELISA-based Met biochemical assay using the random copolymer poly(Glu:Tyr) (4:1) immobilized on microtiter plates as a substrate. IC50 value is determined with various concentrations of SU11274 in a buffer containing 5 μM ATP and 10 mM MnCl2, 50 mM HEPES (pH 7.5), 25 mM NaCl, 0.01% BSA, and 0.1 mM Na orthovanadate. The kinase reaction is performed for 5 minutes at room temperature. The extent of substrate phosphorylation is measured using horseradish peroxidase-conjugated anti-pTyr antibodies.

Cells are exposed to various concentrations of SU11274 in the presence or absence of HGF for 24, 48, and 72 hours. The number of viable cells is determined using the MTT assay or trypan blue exclusion. Cell Cycle and apoptosis are measured by fluorescence-activated cell sorter analysis via propidium iodide staining and Annexin V-positive staining, respectively. (Only for Reference)