Glumetinib (SCC244) is a novel potent and selective inhibitor of c-Met kinase (IC50: 0.42 nM).

c-Met:0.42 nM (cell free)

Glumetinib在使用ELISA激酶测定法对纯化的c-Met激酶活性表现出高效能(IC50: 0.42 nM)。Glumetinib对c-Met的选择性超过了2,400倍，远高于所评估的312种激酶，包括c-Met家族成员RON以及与之高度同源的激酶Axl、Mer和TyrO3。Glumetinib强烈抑制了HGF诱导的NCI-H441细胞的迁移和侵袭能力，并且在10 nmol/L的剂量下足以阻止大多数细胞的移动，显示出剂量依赖性的作用。

在MKN-45模型中，Glumetinib以10、5和2.5 mg/kg的剂量分别显著抑制肿瘤生长，抑制率分别为99.3%、88.6%和63.6%。此外，在使用5和10 mg/kg剂量的Glumetinib进行21天治疗后，观察到肿瘤停滞。在SNU-5模型中使用Glumetinib治疗得到了类似结果，并且在高剂量组中观察到肿瘤回归。在EBC-1研究中，所有接受Glumetinib治疗的小鼠，其肿瘤质量均减少了超过66.0%，且在10和5 mg/kg治疗组中，每6只小鼠中就有1只未发现肿瘤。此外，在所有测试模型中，Glumetinib在10 mg/kg的效力与INCB28060在15 mg/kg和crizotinib在50 mg/kg的效力相当。

Met, Ron, Axl, TyrO3, and Mer kinases activity were assessed using both ELISA and radiometric protein kinase assays. The kinase selectivity profile of SCC244 (1 μmol/L) was screened against a panel of other 308 recombinant kinases using radiometric protein kinase assays was also performed according to the manufacturer's specifications.

Cells were seeded in 96-well plates at a low density in growth media. The next day, appropriate controls or designated concentrations of compounds were added to each well, and the cells were incubated for 72 hours. HUVECs (passage 3) were seeded in 96-well plates in growth media overnight and transferred to serum-free media for 24 hours. The following day, appropriate controls or designated concentrations of compounds were added to each well, and HGF was added to designated wells at 100 ng/mL. The cells were incubated for 48 hours. Finally, cell proliferation was determined using a sulforhodamine B assay, a thiazolyl blue tetrazolium bromide assay or a cell counting kit (CCK-8) assay.

To assess the pharmacodynamics of SCC244 in tumors, mice bearing established xenograft tumors were treated with a single dose of the compound at 10 or 2.5 mg/kg, and tumors were harvested at several time points. At a designated time following administration, mice were humanely euthanized, and their tumors were resected. The tumors were snap-frozen in liquid nitrogen and then homogenized in 500 μL of protein extraction solution (radioimmunoprecipitation assay, RIPA). The tumor extracts were then subjected to Western blot analysis. The individual bands of phospho-c-Met, phospho-AKT, and phospho-ERK were scanned and quantified using Gel Pro Analyzer software. The relative tyrosine phosphorylation of each sample at the indicated time points was then calculated, with the average value of vehicle-treated sample used as 100%.