CHIR-98014 - GSK-3 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECHIR-98014Cat. No.: HY-13076CAS No.: 252935-94-7分式: CHClNO分量: 486.31作靶點(diǎn): GSK-3作通路: PI3K/Akt/mTOR; Stem Cell/Wnt儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 11 mg/mL (22.62 mM; Need ultra

2、sonic and warming)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.0563 mL 10.2815 mL 20.5630 mL5 mM 0.4113 mL 2.0563 mL 4.1126 mL10 mM 0.2056 mL 1.0282 mL 2.0563 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 CHIR-98014種有效的，細(xì)胞通透的 GSK-3 抑制劑，可抑制 GSK-3 和 GSK-3 的活性，IC50 值分別為0.

3、65 和 0.58 nM；CHIR-98014 對(duì) cdc2 和 erk2 的作較弱。IC50 & Target GSK-3 GSK-3 cdc20.58 nM (IC50) 0.65 nM (IC50) 3700 nM (IC50)體外研究CHIR 98014 inhibits human GSK-3 with Ki value of 0.87 nM. CHIR 98014 causes GS stimulation in CHO-IR1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEcells and rat hepatocytes, wit

4、h EC50s of 106 nM and 107 nM, respectively 1. CHIR-98014 (1 M) reducesthe viability of ES-CCE cells by 52%, with IC50 of 1.1 M. Moreover, CHIR-98014 in combination with CHIR-99021 results in a significant activation of the Wnt/beta-catenin pathway in ES-D3 cells. In CHIR-98014treated cells, the T ge

5、ne expression is induced up to 2,500-fold. CHIR-98014 (1 M) also yields around 50%Brachyury-positive cells, with EC50 of 0.32 M 2. CHIR98014 (10 M) prevents loss of neurites caused by20 M PrP1-30 in cortical and hippocampal neurons, and substantially decreases the amount of dead cells3.體內(nèi)研究 CHIR 980

6、14 (30 mg/kg, i.p.) exhibits a significant reduction in fasting hyperglycemia within 4 h of treatmentand shows improved glucose disposal during an ipGTT in markedly diabetic and insulin-resistant db/db mice1.PROTOCOLKinase Assay 1 Polypropylene 96-well plates are filled with 300 L/well buffer (50 mM

7、 tris HCl, 10 mM MgCl2, 1 mM EGTA, 1mM dithiothreitol, 25 mM -glycerophosphate, 1 mM NaF, 0.01% BSA, pH 7.5) containing kinase, peptidesubstrate, and any activators. Information on the kinase concentration, peptide substrate, and activator forthese assays is as follows: GSK-3 (27 nM, and 0.5 M bioti

8、n-CREB peptide); GSK-3 (29 nM, and 0.5 Mbiotin-CREB peptide); cdc2 (0.8 nM, and 0.5 M biotin histone H1 peptide); erk2 (400 units/mL, and myelinbasic protein-coated Flash Plate); PKC- (1.6 nM, 0.5 M biotin-histone H1 peptide, and 0.1 mg/mLphosphatidylserine + 0.01 mg/mL diglycerides); PKC- (0.1 nM,

9、0.5 M biotin-PKC-86 peptide, and 50 g/mLphosphatidylserine + 5 g/mL diacylglycerol); akt1 (5.55 nM, and 0.5 M biotin phospho-AKT peptide); p70S6 kinase (1.5 nM, and 0.5 M biotin-GGGKRRRLASLRA); p90 RSK2 (0.049 units/mL, and 0.5 M biotin-GGGKRRRLASLRA); c-src (4.1 units/mL, and 0.5 M biotin-KVEKIGEGT

10、YGVVYK); Tie2 (1 g/mL, and 200nM biotin-GGGGAPEDLYKDFLT); flt1 (1.8 nM, and 0.25 M KDRY1175 B91616 biotin- GGGGQDGKDYIVLPI-NH2); KDR (0.95 nM, and 0.25 M KDRY1175 B91616 biotin- GGGGQDGKDYIVLPI-NH2); bFGF receptor tyrosine kinase (RTK; 2 nM, and 0.25 M KDRY1175 B91616biotin-GGGGQDGKDYIVLPI-NH2); IGF

11、1 RTK (1.91 nM, and 1 M biotin-GGGGKKKSPGEYVNIEFG-amide);insulin RTK (using DG44 IR cells); AMP kinase (470 units/mL, 50 M SAMS peptide, and 300 M AMP);pdk1 (0.25 nM, 2.9 nM unactivated Akt, and 20 M each of DOPC and DOPS + 2 M PIP3); CHK1 (1.4 nM,and 0.5 M biotin-cdc25 peptide); CK1- (3 nM, and 0.2

12、 M biotin-peptide); DNA PK (see 31); andphosphatidylinositol (PI) 3-kinase (5 nM, and 2 g/mL PI). Test compounds or controls are added in 3.5 L ofDMSO, followed by 50 L of ATP stock to yield a final concentration of 1 M ATP in all cell-free assays. Afterincubation, triplicate 100-L aliquots are tran

13、sferred to Combiplate eight plates containing 100 L/well 50 MATP and 20 mM EDTA. After 1 h, the wells are rinsed five times with PBS, filled with 200 L of scintillationfluid, sealed, left 30 min, and counted in a scintillation counter. All steps are performed at room temperature.Inhibition is calcul

14、ated as 100% (inhibited no enzyme control)/(DMSO control no enzyme control) 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 The viability of the mouse ES cells is determined after exposure to different concentrations of GSK3 inhibitorsfor

15、 three days using the MTT assay. The decrease of MTT activity is a reliable metabolism-based test forquantifying cell viability; this decrease correlates with the loss of cell viability. 2,000 cells are seededovernight on gelatine-coated 96-well plates in LIF-containing ES cell medium. On the next d

16、ay the medium is2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEchanged to medium devoid of LIF and with reduced serum and supplemented with 0.1-1 M BIO, or 1-10 MSB-216763, CHIR-99021 or CHIR-98014. Basal medium without GSK3 inhibitors or DMSO is used ascontrol. All tested conditions are analyzed

17、in triplicates 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Blood is obtained by shallow tail snipping at lidocaine-anesthetized tips. Blood glucose is measured directlyAdministration 1 or heparinized plasma is collected for measurement of g

18、lucose or insulin. Animals are prebled andrandomized to vehicle control or GSK-3 inhibitor treatment groups. For glucose tolerance tests (GTTs),animals are fasted throughout the procedure with food removal early in the morning, 3 h before first prebleed(db/db mice), or the previous night, 16 h befor

19、e the bleed (ZDF rats). When the time course of plasmaglucose and insulin changes in fasting ZDF rats is measured, food is removed -16 h before test agentadministration. The glucose challenges in the GTT are 1.35 g/kg i.p. (ipGTT) or 2 g/kg via oral gavage(oGTT). Test inhibitors are formulated as so

20、lutions in 20 mM citrate-buffered 15% Captisol or as finesuspensions in 0.5% carboxymethylcellulose 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Cancer Res. 2019 Feb 1;79(3):534-545.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Ring DB, et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation o