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Research Area
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Description
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Cancer |
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Biological Activity
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Description
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SU11274 is a selective Met inhibitor with IC50 of 10 nM. |
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Targets
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Met |
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IC50 |
10 nM [1] |
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In Vitro
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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] |
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In Vivo
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Clinical Trials
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Features
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Protocol
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Kinase Assay
[1]
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| 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. |
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Cell Assay
[2]
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| 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. |
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References |
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[1] Wang X, et al. Mol Cancer Ther, 2003, 2(11):1085-1092.
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[2] Sattler M, et al. Cancer Res, 2003, 63(17), 5462-5469.
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[3] Ma PC, et al. Cancer Res, 2005, 65(4), 1479-1488.
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