1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECCG215022Cat. No.: HY-18991CAS No.: 1813527-81-9分式: CHFNO分量: 499.5作靶點: PKA作通路: Protein Tyrosine Kinase/RTK; Stem Cell/Wnt儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數據體外實驗 DMSO : 28 mg/mL (56.06 mM)*

2、 means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.0020 mL 10.0100 mL 20.0200 mL5 mM 0.4004 mL 2.0020 mL 4.0040 mL10 mM 0.2002 mL 1.0010 mL 2.0020 mL請根據產品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 CCG215022種G蛋偶聯(lián)受體激酶 (GRK) 抑制劑，作于 GRK2，GRK5 和

3、 GRK1，IC50 分別為0.150.07 M，0.380.06 M 和 3.91 M。IC50 & Target IC50 & Target: 3.91.0 M (GRK1), 0.150.07 M (GRK2), 0.380.06 M (GRK5), 12040 M (PKA) 1體外研究CCG215022 has nanomolar potency against both GRK2 and GRK5 and is at least 20-fold more potent than1/2 Master of Small Molecules 您邊的抑制劑師www.MedChemEParo

4、xetine. In the course of a GRK2 structure-based drug design campaign, CCG215022 exhibits nanomolarIC50 values against both GRK2 and GRK5 and good selectivity against other closely related kinases such asGRK1 and PKA. Treatment of murine cardiomyocytes with CCG215022 results in significantly increase

5、scontractility at 20-fold lower concentrations than Paroxetine, an inhibitor with more modest selectivity forGRK2 1.PROTOCOLKinase Assay 1 GRK5 and urea-washed bovine rod outer segments (ROS) are mixed in the dark in buffer containing 20 mMHEPES, pH 7.5, 4 mM MgCl2, and 2 mM EDTA and incubated for 3

6、5 min at room temperature. The reactionmixtures are exposed to ambient fluorescent light for 1 min prior to initiation of the reaction by addition ofATP (with -32PATP) to a final concentration of 1 mM. Final concentration of GRK5 is 100 nM and ROS isbetween 0.75 and 24 M. Reactions are initiated at

7、room temperature, and samples are taken at 2-5 min andthen quenched with SDSloading dye. Proteins are separated using SDS, gel is dried, and theincorporation of -32P is detected using a phosphor storage screen. Rates at 0 min are plotted against theROS concentration, and Vmax and Kmvalues are determ

8、ined using the Michaelis-Menten equation. Vmax ofeach curve is normalized to the Vmax of GRK5561 run in parallel. Melting point determinations in responseto 200 M CCG215022 are performed in 20 mM HEPES, pH 7.0, 5 mM MgCl2, 2 mM DTT, 1 mM CHAPS ata final GRK5 concentration of 0.2 mg/mL and 100 M anil

9、inonaphthalene-8-sulfonic acid using aThermoFluor plate reader. Melting points of GRK5 variants are assayed in a buffer containing 20 mMHEPES, pH 8.0, 200 mM NaCl, 2 mM DTT, 2.5 mM MgCl2, and 0.1 mM anilinonaphthalene-8-sulfonic acidwith or without 5 mM ATP. Final GRK5 concentration for these assays

10、 is 0.1 mg/mL 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Homan KT, et al. Crystal Structure of G Protein-coupled Receptor Kinase 5 in Complex with a Rationally DesignedInhibitor. J Biol Chem.2015 Aug 21;290(34):20649-59.McePdfHeightCaution: Product has not been fully valid