Probing of exosites leads to novel inhibitor scaffolds of HCV NS3/4A proteinase.
Probing of exosites leads to novel inhibitor scaffolds of HCV NS3/4A proteinase.
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DOI：
10.1371/journal.pone.0040029
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Strongin AY
中科院分区：
文献类型：
--
作者：
Shiryaev SA;Cheltsov AV;Strongin AY
丙型肝炎是一种难治性疾病，影响着全球数百万人。丙型肝炎病毒（HCV）基因组是单链RNA分子。感染宿主细胞后，病毒RNA被翻译成多蛋白，该多蛋白被宿主和病毒蛋白酶切割成功能性、结构性和非结构性病毒蛋白。多蛋白的切割涉及病毒NS 3/4A蛋白酶，这是一种已证实的药物靶点。HCV在复制时会发生突变，因此，多种新出现的准种会迅速对抗病毒药物（包括NS 3/4A抑制剂）产生耐药性。为了避免耐药性并补充现有的抗病毒药物，需要额外的NS 3/4A抑制剂，其不同于FDA批准的特拉匹韦和博赛匹韦α-酮酰胺抑制剂。为了测试抑制剂开发的潜在新途径，我们探测了NS 3/4A的几个不同的外位点，这些外位点在蛋白酶的活性位点沟之外或与之部分重叠。为此，我们采用虚拟配体筛选，使用开发治疗计划（NCI/NIH）的275，000化合物库和NS 3/4A的X射线晶体结构分别作为配体源和靶。因此，我们确定了几种新的，以前未表征的，纳摩尔范围的抑制性支架，抑制NS 3/4A的体外活性和复制的亚基因组HCV RNA复制子与荧光素酶报告在人肝癌细胞。这些新型抑制剂的结合位点与α-酮酰胺的结合位点没有显著重叠。因此，最常见的耐药突变，包括V36 M、R155 K、A156 T、D168 A和V170 A，并没有显著降低我们鉴定的某些新型抑制剂支架的抑制效力。总的来说，我们开发的计算机策略和软件平台以及我们确定的先导化合物的进一步优化可能会导致新型抗病毒药物的进步。
Hepatitis C is a treatment-resistant disease affecting millions of people worldwide. The hepatitis C virus (HCV) genome is a single-stranded RNA molecule. After infection of the host cell, viral RNA is translated into a polyprotein that is cleaved by host and viral proteinases into functional, structural and non-structural, viral proteins. Cleavage of the polyprotein involves the viral NS3/4A proteinase, a proven drug target. HCV mutates as it replicates and, as a result, multiple emerging quasispecies become rapidly resistant to anti-virals, including NS3/4A inhibitors. To circumvent drug resistance and complement the existing anti-virals, NS3/4A inhibitors, which are additional and distinct from the FDA-approved telaprevir and boceprevir α-ketoamide inhibitors, are required. To test potential new avenues for inhibitor development, we have probed several distinct exosites of NS3/4A which are either outside of or partially overlapping with the active site groove of the proteinase. For this purpose, we employed virtual ligand screening using the 275,000 compound library of the Developmental Therapeutics Program (NCI/NIH) and the X-ray crystal structure of NS3/4A as a ligand source and a target, respectively. As a result, we identified several novel, previously uncharacterized, nanomolar range inhibitory scaffolds, which suppressed of the NS3/4A activity in vitro and replication of a sub-genomic HCV RNA replicon with a luciferase reporter in human hepatocarcinoma cells. The binding sites of these novel inhibitors do not significantly overlap with those of α-ketoamides. As a result, the most common resistant mutations, including V36M, R155K, A156T, D168A and V170A, did not considerably diminish the inhibitory potency of certain novel inhibitor scaffolds we identified. Overall, the further optimization of both the in silico strategy and software platform we developed and lead compounds we identified may lead to advances in novel anti-virals.
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影响因子：
13.5
作者：
Kieffer, Tara L.;Sarrazin, Christoph;Zeuzem, Stefan
通讯作者：
Zeuzem, Stefan
DOI：
10.1016/j.bbapap.2009.06.004
发表时间：
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影响因子：
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作者：
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通讯作者：
Allen, Todd M.
影响因子：
3.7
作者：
Cubero, Maria;Esteban, Juan Ignacio;Quer, Josep
通讯作者：
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影响因子：
29.4
作者：
Hinrichsen, H;Benhamou, Y;Steinmann, GG
通讯作者：
Steinmann, GG