Identification and analysis of hepatitis C virus NS3 helicase inhibitors using nucleic acid binding assays.
Identification and analysis of hepatitis C virus NS3 helicase inhibitors using nucleic acid binding assays.
复制标题
DOI：
10.1093/nar/gks623
复制
发表时间：
2012-09-01
影响因子：
14.9
通讯作者：
Frick DN
中科院分区：
文献类型：
--
作者：
Mukherjee S;Hanson AM;Shadrick WR;Ndjomou J;Sweeney NL;Hernandez JJ;Bartczak D;Li K;Frankowski KJ;Heck JA;Arnold LA;Schoenen FJ;Frick DN
用于发现和分析抑制丙型肝炎病毒（HCV）NS 3解旋酶的小分子的典型测定产生很少的命中，并且经常被化合物干扰混淆。寡核苷酸结合试验在这里检查作为一种替代。在比较荧光偏振（FP）、均匀时间分辨荧光（HTRF®; Cisbio）和AlphaScreen®（Perkin Elmer）测定之后，选择基于FP的测定来筛选Sigma的药理学活性化合物文库（LOPAC）中抑制NS 3-DNA复合物形成的化合物。四种LOPAC化合物抑制基于FP的测定：金精三羧酸（ATA）（IC 50 = 1.4 μM）、苏拉明钠盐（IC 50 = 3.6 μM）、NF 023水合物（IC 50 = 6.2 μM）和tyrphostin AG 538（IC 50 = 3.6 μM）。除AG 538外，所有的抑制解旋酶催化的链分离，除NF 023外，所有的抑制亚基因组HCV复制子的复制。使用大肠杆菌单链DNA结合蛋白（SSB）的反筛选显示，没有一种新的HCV解旋酶抑制剂对NS 3 h具有特异性。然而，当基于SSB的测定用于分析另一种非特异性解旋酶抑制剂（染料报春碱的主要组分）的衍生物时，其揭示了一些报春碱衍生物（例如PubChem CID 50930730）对HCV NS 3 h的特异性比类似有效的HCV解旋酶抑制剂高30倍。
Typical assays used to discover and analyze small molecules that inhibit the hepatitis C virus (HCV) NS3 helicase yield few hits and are often confounded by compound interference. Oligonucleotide binding assays are examined here as an alternative. After comparing fluorescence polarization (FP), homogeneous time-resolved fluorescence (HTRF®; Cisbio) and AlphaScreen® (Perkin Elmer) assays, an FP-based assay was chosen to screen Sigma’s Library of Pharmacologically Active Compounds (LOPAC) for compounds that inhibit NS3-DNA complex formation. Four LOPAC compounds inhibited the FP-based assay: aurintricarboxylic acid (ATA) (IC50 = 1.4 μM), suramin sodium salt (IC50 = 3.6 μM), NF 023 hydrate (IC50 = 6.2 μM) and tyrphostin AG 538 (IC50 = 3.6 μM). All but AG 538 inhibited helicase-catalyzed strand separation, and all but NF 023 inhibited replication of subgenomic HCV replicons. A counterscreen using Escherichia coli single-stranded DNA binding protein (SSB) revealed that none of the new HCV helicase inhibitors were specific for NS3h. However, when the SSB-based assay was used to analyze derivatives of another non-specific helicase inhibitor, the main component of the dye primuline, it revealed that some primuline derivatives (e.g. PubChem CID50930730) are up to 30-fold more specific for HCV NS3h than similarly potent HCV helicase inhibitors.
登录
查看更多内容
影响因子：
3.7
作者：
Hashem AM;Flaman AS;Farnsworth A;Brown EG;Van Domselaar G;He R;Li X
通讯作者：
Li X
DOI：
10.1038/nrd1853
发表时间：
2005-10
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Kwong AD;Rao BG;Jeang KT
通讯作者：
Jeang KT
影响因子：
5.7
作者：
Kim, JL;Morgenstern, KA;Caron, PR
通讯作者：
Caron, PR
影响因子：
2.7
作者：
Gemma, Sandra;Butini, Stefania;Maga, Giovanni
通讯作者：
Maga, Giovanni
影响因子：
--
作者：
Chen, Ye;Bopda-Waffo, Alain;Kaushik-Basu, Neerja
通讯作者：
Kaushik-Basu, Neerja