SB1317 (TG02) is a potent inhibitor of cyclin dependant kinases (CDKs), Janus kinase 2 (JAK2), and Fms-like tyrosine kinase-3 (FLT3).

SB1317是一种具有高度独特的激酶抑制谱的化合物，能够抑制一个由63个激酶组成的面板中的17种激酶，其中11种属于CDK/JAK/FLT家族。其余的，包括Lck、Fyn、Fms、TYRO3、ERK5和p38δ，都与炎症和增殖过程有关。在最高测试浓度25 μM时，人类CYP1A2、3A4、2C9和2C19同工酶未被SB1317抑制，但SB1317以IC50=0.95 μM抑制CYP2D6，接近于最大耐受剂量时观察到的血浆Cmax。SB1317在HCT-116（IC50=0.079 μM）和HL-60（IC50=0.059 μM）细胞中抑制细胞增殖[1]。SB1317是一种新型的、小分子的强效CDK/JAK2/FLT3抑制剂，主要通过CYP3A4和CYP1A2在体外代谢。除CYP2D6（IC50=1 μM）外，SB1317不抑制任何主要的人类CYPs，并且在体外的人类肝细胞中不显著诱导CYP1A和CYP3A4[2]。

The recombinant enzymes (CDK2/cyclin A, JAK2, and FLT3) are used. All assays are carried out in 384-well white microtiter plates using the PKLight assay system. This assay platform is a luminometric assay for the detection of ATP in the reaction using a luciferase-coupled reaction. The compounds are tested at eight concentrations prepared from 3- or 4-fold serial dilution starting at 10 μM. For CDK2/cyclin A assay, the reaction mixture consisted of the following components in 25 μL of assay buffer (50 mM Hepes, pH 7.5, 10 mM MgCl2, 5 mM MnCl2, 5 mM BGP, 1 mM DTT, 0.1 mM sodium orthovanadate), 1.4 μg/mL of CDK2/cyclin A complex, 0.5 μM RbING substrate, and 0.5 μM ATP. The mixture is incubated at room temperature for 2 h. Then 13 μL of PKLight ATP detection reagent is added and the mixture is incubated for 10 min. Luminescence signals are detected on a multilabel plate reader. The analytical software Prism 5.0 is used to generate IC50 values from the data[1].

SB1317 is prepared in DMSO and stored, and then diluted with appropriate medium before use[1]. All cell lines are obtained from the American Type Culture Collection and cultured. For proliferation assays in 96-well plates, 20 000 cells are seeded in 100 μL of medium and treated the following day with compounds (e.g., SB1317 ) (in triplicate) at concentrations up to 10 μM for 48 h. Cell viability is monitored using the CellTiter-96 Aqueous One solution cell proliferation assay. Dose-response curves are plotted to determine IC50 values for the compounds using the XL-fit software[1].