PSI-6206_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries
Data Sheet
BIOLOGICAL ACTIVITY:
PSI–6206 is the deaminated derivative of PSI–6130, which is a potent and selective inhibitor of HCV NS5B polymerase. PSI–6206 low potently inhibits HCV replicon with EC 90 of >100 μM.
IC50 & Target: EC90:>100 μM (HCV replicon)[1]
In Vitro: PSI–6206 (RO2433) is tested for anti–HCV activity in both a cell–based quantitative real–time RT–PCR assay and surrogate bovine viral diarrhea virus (BVDV) assays. PSI–6206 demonstrates no activity or cytoxicity in any assay [1]. The formation of the
5′–triphosphate (TP) of PSI–6130 (PSI–6130–TP) and RO2433 (RO2433–TP) increases with time and reached steady state levels at 48h. RO2433–TP also inhibits RNA synthesis by the native HCV replicase isolated from HCV replicon cells and the recombinant HCV polymerase NS5B [2]. PSI–6206 (RO2433) is the deaminated derivative of PSI–6130, which is a potent and selective inhibitor of HCV NS5B polymerase [3].
PROTOCOL (Extracted from published papers and Only for reference)
Enzyme assay [3]
The DCTD assays were performed by using a spectrophotometric assay for the natural substrate, dCMP, and an HPLC assay for PSI–6130–MP. The spectrophotometric assay was essentially the same as the CDA assay described above. The reactions were
performed at 37 °C, and the change in absorbance at 290 nm (Δ = 1,694 M1 cm1) was monitored. The final enzyme concentration was
9.3 nM. The reactions for the HPLC assay were initiated by adding 0.56 μg of enzyme to a reaction mixture (500 μl) containing 50 mM Tris–HCl, pH 7.5, 2 mM MgCl2, 20 mM β–mercaptoethanol, and the substrate at the indicated concentrations. The reaction mixtures were incubated at 37°C; and at 5, 10, and 20 min, 100 μl of the reaction mixture was removed and quenched with 3.5 to 3.8% (wt/wt)HCl. Prior to HPLC analysis, each quenched sample was passed through a YM–10 Microcon centrifugal filter unit (Millipore), stored at ?20°C, and analyzed within 24 h with a Perkin–Elmer (Waltham, MA) series 200 HPLC system. Strong–anion–exchange HPLC was performed on a Whatman 10–μm Partisil SAX column (Whatman, Maidstone, England) which was preequilibrated with buffer A (0.02M KH2PO4, pH 3.5). Elution of the nucleotides was accomplished with a 10–min linear gradient with from 0 to 10% buffer B (1 M KH2PO4, pH 3.5) at a flow rate of 1.0 ml/min. The nucleotide substrate and product were detected at 254 nm and were quantified on the basis of a standard curve of the peak area versus the concentration.
References:
[1]. Clark JL, et al. Design, Synthesis, and Antiviral Activity of 2'–Deoxy–2'–fluoro–2'–C–methyl–cytidine, a Potent Inhibitor of Hepatitis C Virus Replication. J Med Chem. 2005 Aug 25;48(17):5504–8.
[2]. Ma H, et al. Characterization of the metabolic activation of hepatitis C virus nucleoside inhibitor beta–D–2'–Deoxy–2'–fluoro–2'–C–methylcytidine (PSI–6130) and identification of a novel active 5'–triphosphate species. J Biol Chem. 2007 Oct 12;282(41):29
Product Name:
PSI–6206Cat. No.:
HY-15236CAS No.:
863329-66-2Molecular Formula:
C 10H 13FN 2O 5Molecular Weight:
260.22Target:
HCV Pathway:
Anti–infection Solubility:
10 mM in DMSO
[3]. Wang P, et al. An efficient and diastereoselective synthesis of PSI–6130: a clinically efficacious inhibitor of HCV NS5B polymerase. J Org Chem. 2009 Sep 4;74(17):6819–24.
Caution: Product has not been fully validated for medical applications. For research use only.
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