BX-912 - PDK-1 抑制剂 - 生命科学试剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEBX-912Cat. No.: HY-11005CAS No.: 702674-56-4分式: CHBrNO分量: 471.35作靶点: PDK-1作通路: PI3K/Akt/mTOR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 (212.16 mM)* means soluble, but saturat

2、ion unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.1216 mL 10.6078 mL 21.2157 mL5 mM 0.4243 mL 2.1216 mL 4.2431 mL10 mM 0.2122 mL 1.0608 mL 2.1216 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，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

3、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: 2.75 mg/mL (5.83 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.75 mg/mL (5.83 mM); Clear solution3. 请依序添加每种溶剂： 10% DMSO 90% corn oil1/3 Master of Small Molecules

4、您边的抑制剂师www.MedChemESolubility: 2.75 mg/mL (5.83 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 BX-912种有效的 PDK1 抑制剂，IC50 为 12 nM。IC50 & Target IC50: 12 nM (PDK1), 110 nM (PKA), 410 nM (KDR), 650 nM (CDK2/cyclin E), 830 nM (Chck1), 850 nM (c-Kit), 1.25 M (PKC), 2.1 M (T-Fyn), 6.1 M (Insulin receptor), 7.4

5、M (GSK3) 1体外研究 BX-912 also inhibits PKA, KDR, CDK2/cyclin E, Chck1, c-Kit, PKC, T-Fyn, Insulin receptor, and GSK3 withIC50s of 110 nM, 410 nM, 650 nM, 830 nM, 850 nM, 1.25 M, 2.1 M, 6.1 M and 7.4 M in kinase assays,respectively. BX-912 is identified in a coupled assay measuring PDK1- and PtdIns-3,4-

6、P2-mediated Aktactivation, which can detect inhibitors of PDK1, AKT2, or other steps critical for activation of AKT2. BX-912potently inhibits PDK1 enzyme activity in a direct kinase assay format (IC50=26), although BX-912 fails toblock preactivated AKT2 activity (IC5010 M). BX-912 binds to the ATP b

7、inding site of PDK1. Theaminopyrimidine backbone of BX-912 adopts a similar orientation in the active site of PDK1. To examine thekinase selectivity of BX-912, its effects on the in vitro activity of 10 different Ser/Thr and tyrosine kinases aredetermined, including the related AGC kinases PKA and P

8、KC. BX-912 is 9-fold selective for PDK1 relativeto PKA. BX-912 blocks PDK1 activity in PTEN-negative PC-3 cells. PTEN-negative PC-3 cells displayconstitutive activation of Akt which is reflected in high levels of the PDK1 product, phospho-Thr308-Akt 1.PROTOCOLKinase Assay 1 PDK1 is assayed in a dire

9、ct kinase assay and a coupled assay format measuring PDK1 and PtdIns-3,4-P2mediated activation of AKT2. For the coupled assay, the final assay mixture (60 L) contains: 15 mM MOPS,pH 7.2, 1 mg/mL bovine serum albumin, 18 mM -glycerol phosphate, 0.7 mM dithiothreitol, 3 mM EGTA, 10mM MgOAc, 7.5 M ATP,

10、 0.2 Ci of -33PATP, 7.5 M biotinylated peptide substrate (biotin-ARRRDGGGAQPFRPRAATF), 0.5 L of PtdIns-3,4-P2-containing phospholipid vesicles, 60 pg of purifiedrecombinant human PDK1, and 172 ng of purified recombinant human AKT2. After incubation for 2 h at roomtemperature, the biotin-labeled pept

11、ide is captured from 10 L of the assay mixture on Streptavidin-coatedSPA beads, and product formation is measured by scintillation proximity in a Wallac MicroBeta counter. Theproduct formed is proportional to the time of incubation and to the amount of PDK1 and inactive AKT2 added.PDK1 is added at s

12、uboptimal levels so that the assay can sensitively detect inhibitors of AKT2 activation aswell as direct inhibitors of PDK1 or AKT2 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The cell lines MDA-468, MDA-453, HCT-116, U87-MG, U2OS, PC

13、-3, B16F10, and MiaPaCa; LOXamelanotic human melanoma cells; and HeLa cells seeded at a low density (1,500-3,000 cells/well, 0.1mL/well, 96-well plates) are incubated overnight. Compound treatments are made by adding 10 L/well ofBX-912 (1, 10, 100 and 1000 nM) in 1% DMSO and growth medium (final con

14、centration of DMSO, 0.1%),followed by brief shaking. Treated cells are incubated for 72 h, and viability is measured by the addition of 10L of the metabolic dye WST-1. The WST-1 signal is read in a plate reader at 450 nm, and a no cell, or zero2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEtime ce

15、ll, background is subtracted to calculate the net signal 1.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 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Cell Syst. 2018 Dec. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Feldman RI, et al. Novel small molecule inhibitors of 3-phosphoinositide-dependent kinase-1. J Biol Che