CVT-313-Cdk2-Inhibitor-III-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECVT-313Cat. No.: HY-15339CAS No.: 199986-75-9Synonyms: Cdk2 Inhibitor III分式: CHNO分量: 400.47作靶点: CDK作通路: Cell Cycle/DNA Damage储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 100 mg/mL (249.7

2、1 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.4971 mL 12.4853 mL 24.9707 mL5 mM 0.4994 mL 2.4971 mL 4.9941 mL10 mM 0.2497 mL 1.2485 mL 2.4971 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性

3、，体内实验的作液，建议您现现配，当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (6.24 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (6.24 mM); Clear solution1/3 Master of Small Molecules 您边的抑

4、制剂师www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (6.24 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 CVT-313种有效的ATP-竞争性的选择性 CDK2 抑制剂，IC50 为 0.5 M。IC50 & Target cdk2/cyclin A Cdk1/cyclin B Cdk4/cyclin D10.5 M (IC50) 4.2 M (IC50) 215 M (IC50)体外研究 CVT-313 has been shown to inhibit ot

5、her kinases, but at much higher IC50 values, i.e., CDK1 (IC50=4.2 M),CDK4 D1 (IC50=215 M), and MAPK/PKA/PKC (IC501.25 mM), compared to CDK2 (IC50=0.5 M). CVT-313 has been shown to have profound effects on cell proliferation at concentrations of 5-20 M 1. CVT-313is a potent CDK2 inhibitor, which is i

6、dentified from a purine analog library with an IC50 of 0.5 M in vitro.Inhibition is competitive with respect to ATP (Ki=95 nM), and selective CVT-313 has no effect on other,nonrelated ATP-dependent serine/threonine kinases. When added to CDK1 or CDK4, a 8.5- and 430-foldhigher concentration of CVT-3

7、13 is required for half-maximal inhibition of the enzyme activity. Using normaland tumor human/murine cell lines, the effects of CVT-313 on cell proliferation is measured. The IC50 forgrowth inhibition ranged from 1.25 to 20 M 2.PROTOCOLKinase Assay 1 For kinase assays, purified CDC5L(295-795)-His6

8、is mixed with -32PATP, COS-7 cell extract, andincubated in 100 L 20 mM HEPES, pH 7.5, 50 mM NaCl, 2 mM MnCl2, 10 mM MgCl2, 0.5% NP-40, 0.5 mMPMSF, 5 mM benzamidine hydrochloride, 5 mM NaF, 1 mM NaVO3 and the specific inhibitor at 30C for 10minutes. Cell extract as a source of kinase activity is prep

9、ared from subconfluent, serum-stimulated COS-7cells lysed in 20 mM HEPES-NaOH, pH 7.5, 50 mM NaCl, 1% Triton X-100, 10% glycerol, protease andphosphotase inhibitors. Phosphorylated proteins are separated by electrophoresis in 15% polyacrylamide-SDS gels. Specific inhibitors included 20 M staurospori

10、ne, 10 M genistein, 1 M CVT-313, 10 M Rp-MB-cAMPS and 50 M PD98059 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 MRC-5 cells are grown in Dulbeccos modified Eagles medium containing 5% fetal calf serum. CVT313 (0, 5,10, 15 M) is added t

11、o exponentially growing cells in tissue culture. Cell population is measured. Proliferationassays are carried out using the nonradioactive CellTiter 96 kit after 48-h exposure. For FACS analysis ofDNA content, cells are trypsinized, fixed in 70% ice-cold ethanol, and treated with 0.1 mg/mL RNase A a

12、nd40 g/mL propidium iodide for 1 h at 37C 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Biomed Res Int. 2019 May 16;2019:2821731.2/3 Master of Small Molecules 您边的抑制剂师www.MedChemESee more customer validations on HYPERLINK / www.MedChemERE

13、FERENCES1. Graub R, et al. Cell cycle-dependent phosphorylation of human CDC5 regulates RNA processing. Cell Cycle. 2008 Jun 15;7(12):1795-803.2. Brooks EE, et al. CVT-313, a specific and potent inhibitor of CDK2 that prevents neointimal proliferation. J Biol Chem. 1997 Nov14;272(46):29207-11.McePdfH