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

VEGFR2:32 nM

在4T1和CT26肿瘤模型中,SKLB1002和局部热疗产生协同抗血管生成,抗癌和促进细胞凋亡功效.在携带SW620或HepG2异种移植物的无胸腺小鼠中,SKLB1002（100 mg/kg每天,i.p.）引起显著的肿瘤生长抑制,抑制肿瘤血管生成并诱导肿瘤细胞凋亡.SKLB1002在斑马鱼胚胎中,明显阻断胚胎形成和肿瘤诱导的血管生成,对正常细胞增殖没有影响或影响很小.

SKLB1002通过抑制VEGF诱导的VEGFR2激酶磷酸化和下游蛋白激酶,包括ERK,FAK和Src,明显抑制人脐静脉内皮细胞增殖,迁移,侵袭和管腔形成。SKLB1002对正常人细胞L-02细胞毒性显著降低。

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)