Description

Cancer

Description

SU11274 is a selective Met inhibitor with IC50 of 10 nM.

Targets

IC₅₀

In Vitro

SU11274 exhibits greater than 50-fold selectivity for Met versus Flk and more than 500 times selectivity versus other tyrosine kinases such as FGFR-1, c-src, PDGFbR, and EGFR. SU11274 inhibits the phosphorylation of key regulators of the PI3K pathway, including AKT, FKHR, or GSK3β. SU11274 treatment inhibits the growth of TPR-MET-transformed BaF3 cells in a dose-dependent manner with IC50 of <3 μm="" in="" the="" absence="" of="" interleukin="" 3,="" without="" growth="" inhibition="" of="" baf3="" cells="" transformed="" by="" other="" oncogenic="" tyrosine="" kinases,="" including="" bcr-abl,="" tel-jak2,="" tel-abl,="" and="" tel-pdgfβr.="" in="" addition="" to="" cell="" growth,="" su11274="" treatment="" significantly="" inhibits="" the="" migration="" of="" baf3.="" tpr-met="" cells="" by="" 44.8%="" and="" 80%="" at="" 1="" μm="" and="" 5="" μm,="" respectively.="" su11274="" inhibits="" hgf-dependent="" phosphorylation="" of="" met="" as="" well="" as="" hgf-dependent="" cell="" proliferation="" and="" motility="" with="" an="" ic50="" of="" 1-1.5="" μm.="" in="" h69="" and="" h345="" cells="" which="" have="" functional="" met="" receptor,="" su11274="" inhibits="" the="" hgf-induced="" cell="" growth="" with="" ic50="" of="" 3.4="" μm="" and="" 6.5="" μm,="" respectively.="" su11274="" induces="" g1="" cell="" cycle="" arrest="" with="" cells="" in="" g1="" phase="" increased="" from="" 42.4%="" to="" 70.6%="" at="" 5="" μm,="" and="" induces="" caspase-dependent="" apoptosis="" by="" 24%="" at="" 1="" μm.="">^[2] SU11274 inhibits cell viability in c-Met-expressing non-small cell lung cancer (NSCLC) cells with IC50 values of 0.8-4.4 μM, and abrogates hepatocyte growth factor-induced phosphorylation of c-Met and its downstream signaling. ^[3]

In Vivo

Clinical Trials

Features

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.

Cell Lines

BaF3.TPR-MET, H69 and H345 cells

Concentrations

Dissolved in DMSO, final concentrations ~10 μM

Incubation Time

24, 48, and 72 hours

Methods

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.