PSI-6206 (RO 2433) (RO2433) is a selective HCV RNA polymerase inhibitor.

PSI-6206 (RO2433) 在一个基于细胞的定量实时RT-PCR分析和替代性牛病毒性腹泻病毒(BVDV)分析中被测试其抗HCV活性，结果显示在任何分析中均未展现出活性或细胞毒性[1]。PSI-6130 (PSI-6130-TP) 与 RO2433 (RO2433-TP) 的5′-三磷酸（TP）形成随时间增加，并在48小时达到稳态水平。RO2433-TP 还能抑制由HCV复制子细胞分离出的原生HCV复制酶以及重组HCV聚合酶NS5B的RNA合成[2]。PSI-6206 (RO2433) 是PSI-6130的脱氨基衍生物，PSI-6130是一种强效且选择性的HCV NS5B聚合酶抑制剂[3]。

Protein Kinase Assays: The in vitro kinase assays are performed in 96-well plates (30 μL) at ambient temperature for 15–45 min using the recombinant glutathione S-transferase-fused kinase domains (4-100 ng, depending on specific activity). [γ33P]ATP is used as phosphate donor and polyGluTyr-(4:1) peptide as acceptor. With the exception of protein kinase C-α, cyclin-dependent kinase 1/cycB and protein kinase A are protamine sulfate (200 μg/mL), histone H1 (100 μg/mL), and the heptapeptide Leu-Arg-Arg-Ala-Ser-Leu-Gly (known as Kemptide Bachem) respectively and are used as peptide substrates. Assays are optimized for each kinase using the following ATP concentrations: 1.0 μM (c-Kit, c-Met, c-Fms, c-Raf-1, and RET), 2.0 μM (EGFR, erbB2, ErbB3, and ErbB4), 5.0 μM (c-abl), 8.0 μM (Flt-1, Flt-3, Flt-4, Flk, KDR, FGFR-1, and Tek), 10.0 μM (PDGFR-β, protein kinase C-α, and cyclin-dependent kinase 1), and 20.0 μM (c-Src and protein kinase A). The reaction is terminated by the addition of 20 μL 125 mM EDTA. Thirty μL (c-abl, c-Src, insulin-like growth factor-1R, RET-Men2A, and RET-Men2B) or 40 μL (all other kinases) of the reaction mixture is transferred onto Immobilon-polyvinylidene difluoride membrane, presoaked with 0.5% H3PO4 and mounted on a vacuum manifold. Vacuum is then applied and each well rinsed with 200 μL 0.5% H3PO4. Membranes are removed and washed four times with 1.0% H3PO4 and once with ethanol. Dried membranes are counted after mounting in a Packard TopCount 96-well frame and with the addition of 10 μL/well of Microscint. IC50 values (±SE) are calculated by linear regression analysis of the percentage inhibition and are averages of at least three determinations.