PSI-7409-tetrasodium-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPSI-7409 tetrasodiumCat. No.: HY-15745ACAS No.: 1621884-22-7分式: CHFNNaOP分量: 592.12作靶點: HCV作通路: Anti-infection儲存式: Powder -20C 3 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 H2O : 125 mg/mL (211.11 mM)* means soluble, but s

2、aturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 1.6888 mL 8.4442 mL 16.8885 mL5 mM 0.3378 mL 1.6888 mL 3.3777 mL10 mM 0.1689 mL 0.8444 mL 1.6888 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備液，并請注意儲備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 PSI-7409 tetrasodium種 sofosbuvir (PSI-7977) 的 5-三磷酸活性代謝物，能夠抑制 HCV

3、 NS5Bpolymerases，對 GT 1b_Con1，GT 2a_JFH1，GT 3a 和 GT 4a NS5B 聚合酶的 IC50 值分別為 1.6，2.8，0.7 和2.6 M。IC50 & Target IC50: 1.6 M (GT 1b_Con1), 2.8 M (GT 2a_JFH1), 0.7 M (GT 3a), 2.6 M (GT 4a) 1體外研究PSI-7409 tetrasodium is an active 5-triphosphate metabolite, inhibiting HCV NS5B polymerases, with IC50s1/2 Mast

4、er of Small Molecules 您邊的抑制劑師www.MedChemEof 1.6, 2.8, 0.7 and 2.6 M for GT 1b_Con1, GT 2a_JFH1, GT 3a, and GT 4a NS5B polymerases,respectively. PSI-7409 also weakly inhibits human DNA polymerase , with an IC50 of 550 M, but shows noinhibition on DNA Pol and 1. In clone A cells, the levels of PSI-740

5、9 gradually increases to a maximumconcentration of about 25 M over a period of 48 h. PSI-7409 forms at a much faster rate in primary humanhepatocytes, achieving a maximum intracellular concentration of -100 M at 4 h and remains at thatconcentration for 48 h 2.PROTOCOLKinase Assay 1 Human DNA polymer

6、ase , , or is assayed in a 10-L reaction mixture containing 50 mM Tris (pH 7.5), 50mM NaCl, 3 mU/L activated calf thymus DNA, a 20 M concentration of all four natural deoxynucleosidetriphosphates, 4 Ci -32PdCTP, 5 mM MgCl2, and increasing concentrations of PSI-7409 (up to 1 mM), D-ddFCTP, or aphidic

7、olin. DNA polymerase , , or is added to the reaction mixture to give finalconcentrations of 20, 18, and, 50 g/mL, respectively. All reactions are run at 37C and quenched at 30 minby mixing with 1 L of 0.5 M EDTA. The radiolabeled products are quantified. A nonlinear fit is performed todetermine the

8、IC50. The activity of RNA polymerase II is determined in a 25-L in vitro transcription reactionmixture containing 100 ng of cytomegalovirus (CMV) immediate-early promoter DNA, 400 M ATP, CTP, andUTP, 16 M GTP, 10 Ci -32PGTP, 3 mM MgCl2, and various concentrations of PSI-7409 (up to 1 mM),3-dCTP, or

9、-amanitin in transcription buffer (20 mM HEPES pH 7.9, 100 mM KCl, 0.2 mM EDTA, 0.5 mMDTT, and 20% glycerol). All reactions are run at 30C and quenched at 60 min by mixing with 125 L of stopsolution (0.3 M Tris-HCl pH 7.4, 0.3 M sodium acetate, 0.5% SDS, 2 mM EDTA, and 3 g/mL tRNA). TheRNA product i

10、s purified. The resulting samples contains 12 L and the same volume of gel loading dye (98%formamide, 10 mM EDTA, 0.1% xylene cyanol, and 0.1% bromophenol blue) is added. The samples areheated at 90C for 5 min and loaded onto a 6% polyacrylamide sequencing gel. After running, the gel isexposed to a

11、phosphorscreen, and the product is visualized and quantified by using a phosphorimager 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Lam AM, et al. PSI-7851, a pronucleotide of beta-D-2-deoxy-2-fluoro-2-C-methyluridine monophosphate, is a potent and pan-genotypeinhibitor of hepatitis C virus replication. Antimicrob Agents Chemother. 2010 Aug;54(8):3187-96.2. Murakami E, et al. Mechanism of activation of PSI-7851 and its diastereoisomer PSI-7977. J Biol Chem. 2010 Nov 5;285(45):34337-47.McePdfHeightCaution: Pro