RKI-1447 - ROCK 抑制剂 - 生命科学试剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemERKI-1447Cat. No.: HY-15755CAS No.: 1342278-01-6分式: CHNOS分量: 326.37作靶点: ROCK作通路: Cell Cycle/DNA Damage; Stem Cell/Wnt; TGF-beta/Smad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 : 50 mg/mL (

2、153.20 mM)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (7.66 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (7.66 mM); Clear solution1/3 Master of Small Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 RKI-1447效的ROCK1和ROCK2的分抑制剂，IC50值分别为14.5 nM 和6.2 nM

3、。IC50 & Target ROCK2 ROCK16.2 nM (IC50) 14.5 nM (IC50)体外研究 RKI-1447 is a Type I kinase inhibitor that binds the ATP binding site through interactions with the hingeregion and the DFG motif. RKI-1447 suppresses phosphorylation of the ROCK substrates mLC-2 and MYPT-1 in human cancer cells, but has no

4、effect on the phosphorylation levels of the AKT, MEK and S6 kinase atconcentrations as high as 10 M. RKI-1447 is also highly selective at inhibiting ROCK-mediated cytoskeletonre-organization. RKI-1447 inhibits migration, invasion and anchorage-independent tumor growth of breastcancer cells 1.体内研究 RK

5、I-1447 is highly effective at inhibiting the outgrowth of mammary tumors in a transgenic mouse model.RKI-1447 inhibits mammary tumor growth by 87% and on average the mammary tumors from RKI-1447treated mice are 7.7 fold smaller compared to those tumors from mice treated with the vehicle control 1.PR

6、OTOCOLKinase Assay 1 Compounds are tested on three separate days with 8 point dilutions performed in duplicate to determineaverage IC50 values. The assay conditions are optimized to 15 L of kinase reaction volume with 5 ng ofenzyme in 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EGTA, and 0.01% Brij-35.

7、The reaction isincubated for 1 h at room temperature in the presence of 1.5 M of peptide substrate with 12.5 M of ATP(for ROCK1) or 2 M of substrate with 50 M of ATP (for ROCK2). The reaction is then stopped and the ratioof phosphorylated to unphosphorylated peptides is determined by selective cleav

8、age of only theunphosphorylated peptide. The ratio of the signals at 445 nm and 520 nm is measured. IC50 values aredetermined using fitted curves with GraphPad Prism 5 software 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 NIH 3T3 cells

9、 are plated at 8000 cells/well in 8-chamber slides in serum free media for 24 hours, and treatedwith vehicle, 1 M RKI-1447 or 1 M RKI-1313 for 1 hour. The cells are then stimulated with complete media,10 M LPA, 200 ng/mL bradykinin or 30 ng/mL PDGF for 30 mins. After stimulation, the cells are fixed

10、 in 4%paraformaldehyde and stained with Texas-Red Phalloidin. Mounting medium containing DAPI is then addedand at least 100 cells per well are observed 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice: MMTV/neu transgenic mice are treated i

11、.p. daily for 14 days with either Vehicle 20%-2-hydroxypropyl-Administration 1 beta-cyclodextrin (HPCD) or 200 mpk RKI-1447 dissolved in freshly prepared HPCD. The percent change involume is calculated on the basis of the tumor volume on the last day of treatment (Vf) relative to that on theday of i

12、nitiation of treatment (V0). The average percent change in tumor volume is then calculated for eachtreatment group 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEREFERENCES1. Patel RA, et al. RKI-1447 is a potent inhibitor of the Rho-associated ROCK kinases with anti-invasive and antitumor activities in breastcancer. Cancer Res. 2012 Oct 1;72(19):5025-34.McePdfHeightCa