Virtual Ligand Screening of the National Cancer Institute (NCI) Compound Library Leads to the Allosteric Inhibitory Scaffolds of the West Nile Virus NS3 Proteinase
Virtual Ligand Screening of the National Cancer Institute (NCI) Compound Library Leads to the Allosteric Inhibitory Scaffolds of the West Nile Virus NS3 Proteinase
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DOI：
10.1089/adt.2010.0309
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发表时间：
2011-02-01
影响因子：
1.8
通讯作者：
Strongin, Alex Y.
中科院分区：
文献类型：
--
作者：
Shiryaev, Sergey A.;Cheltsov, Anton V.;Strongin, Alex Y.
黄病毒属的病毒是导致重大人类疾病和死亡的原因。黄病毒非结构（NS）3蛋白的N-末端结构域编码丝氨酸胰凝乳蛋白酶折叠蛋白酶（NS 3 pro）。由黄病毒基因组中的上游基因编码的非结构（NS）2B辅因子的存在对于NS 3 pro展现其蛋白水解活性是必需的。NS 2B-NS 3 pro的双组分功能活性对于病毒多蛋白的加工和复制是必需的。NS 2B-NS 3 pro的结构和功能在黄病毒家族中是保守的。由于NS 2B-NS 3 pro在病毒多聚蛋白前体的翻译后加工中的重要功能，它是抗黄病毒药物的一个有前途的靶点。为了鉴定具有降低的交叉反应性和脱靶效应的选择性抑制剂，我们将策略集中在能够靶向NS 2B-NS 3 pro界面而不是NS 3 pro活性位点的变构抑制剂上。使用虚拟配体筛选的多样性，类似于275，000化合物库和催化结构域的双组分西尼罗河病毒（WNV）NS 2B-NS 3 pro作为受体，我们确定了一个有限的子集的新型抑制性支架。几种发现的化合物作为变构抑制剂发挥作用，并在体外切割测定中表现出纳摩尔范围的效力。抑制剂在使用亚基因组、酶标记的WNV和登革病毒复制子的基于细胞的测定中也是有效的。使用弗林蛋白酶（furin）的体外切割测定证实了抑制剂的选择性，弗林蛋白酶是一种人丝氨酸蛋白酶，其底物偏好与WNV NS 2BNS 3 pro的底物偏好相似。从概念上讲，类似的计算机药物发现策略可以很容易地用于鉴定其他黄病毒的抑制剂。
Viruses of the genus Flavivirus are responsible for significant human disease and mortality. The N-terminal domain of the flaviviral nonstructural (NS)3 protein codes for the serine, chymotrypsin-fold proteinase (NS3pro). The presence of the nonstructural (NS)2B cofactor, which is encoded by the upstream gene in the flaviviral genome, is necessary for NS3pro to exhibit its proteolytic activity. The two-component NS2B-NS3pro functional activity is essential for the viral polyprotein processing and replication. Both the structure and the function of NS2B-NS3pro are conserved in the Flavivirus family. Because of its essential function in the posttranslational processing of the viral polyprotein precursor, NS2B-NS3pro is a promising target for anti-flavivirus drugs. To identify selective inhibitors with the reduced cross-reactivity and off-target effects, we focused our strategy on the allosteric inhibitors capable of targeting the NS2B-NS3pro interface rather than the NS3pro active site. Using virtual ligand screening of the diverse, similar to 275,000-compound library and the catalytic domain of the two-component West Nile virus (WNV) NS2B-NS3pro as a receptor, we identified a limited subset of the novel inhibitory scaffolds. Several of the discovered compounds performed as allosteric inhibitors and exhibited a nanomolar range potency in the in vitro cleavage assays. The inhibitors were also potent in cell-based assays employing the sub-genomic, luciferase-tagged WNV and Dengue viral replicons. The selectivity of the inhibitors was confirmed using the in vitro cleavage assays with furin, a human serine proteinase, the substrate preferences of which are similar to those of WNV NS2BNS3pro. Conceptually, the similar in silico drug discovery strategy may be readily employed for the identification of inhibitors of other flaviviruses.