SKLB1002 is a potent and ATP-competitive VEGFR2 inhibitor with IC50 of 32 nM.

VEGFR2:32 nM

SKLB1002 significantly inhibited human umbilical vein endothelial cell proliferation, migration, invasion, and lumen formation by inhibiting VEGF-induced phosphorylation of VEGFR2 kinase and downstream protein kinases, including ERK, FAK, and Src.SKLB1002 significantly reduced cytotoxicity of L-02 in normal human cells.

SKLB1002 significantly inhibited human umbilical vein endothelial cell proliferation, migration, invasion, and lumen formation by inhibiting VEGF-induced phosphorylation of VEGFR2 kinase and downstream protein kinases, including ERK, FAK, and Src.SKLB1002 significantly reduced cytotoxicity of L-02 in normal human cells.

Kinase inhibition assays : Kinase inhibition is measured by the use of radiometric assays conducted by Kinase Profiler service. Briefly, in the presence or absence of SKLB1002, VGFR2 (5–10 mU) is incubated in 25-μL reaction solution containing 8 mmol/L 3-(N-morpholino)propanesulfonic acid (MOPS), pH 7.0, 0.2 mmol/L EDTA, 0.33 mg/mL myelin basic protein, 10 mmol/L Mg acetate, and γ-[33P]ATP. After incubation for 40 minutes at room temperature, the reaction is stopped and 10 μL of the reaction solution is then spotted onto a P30 filtermat and washed 3 times for 5 minutes in 75 mmol/L phosphoric acid and once in methanol prior to scintillation counting.

Cell proliferation is measured using MTT assay. Various cells including HUVECs, L-02, B16-F10, HepG2, and SW620 are treated with indicated concentrations of SKLB1002 for 24 hours. Vandetanib and sunitinib serve as positive controls. Each assay is replicated 3 times.(Only for Reference)