Cinanserin is an inhibitor of the 3C-like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro
Cinanserin is an inhibitor of the 3C-like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro
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DOI：
10.1128/jvi.79.11.7095-7103.2005
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发表时间：
2005-06-01
影响因子：
5.4
通讯作者：
Jiang, HL
中科院分区：
文献类型：
--
作者：
Chen, LL;Gui, CS;Jiang, HL
严重急性呼吸综合征相关冠状病毒（SARS CoV）的3C样蛋白酶（3CL（pro））在病毒的生命周期中起着重要作用，是抗SARS CoV药物最有希望的靶点之一。在这项研究中，一个包含8,000多种现有药物结构信息的数据库通过对接方法进行了虚拟筛选，以确定SARS-CoV 3CL（pro）的潜在结合分子。作为筛选的靶标，使用酶的结合口袋的同源模型和晶体结构。Cinanserin（SQ 10，643）是一种特征良好的5-羟色胺拮抗剂，在20世纪60年代进行了人体初步临床试验，在筛选中显示出较高的评分，并被选择用于进一步的实验评价。用表面等离子体共振技术研究了肉桂色林及其盐酸盐与SARS冠状病毒3CL（pro）及相关的人冠状病毒229 E（HCoV-229 E）的结合。这两种酶的催化活性被抑制，50%抑制浓度（IC 50）值为5 μ M，用荧光底物进行测试。在组织培养试验中进一步评价了桂南色林的抗病毒活性，即基于HCoV-229 E的复制子系统和感染性SARS-CoV和HCoV-229 E的定量试验。所有试验均显示在无毒药物浓度下对冠状病毒复制的强烈抑制作用。病毒RNA和感染性颗粒的水平降低了4个对数单位，IC 50值范围为19至34 μ M。这些发现表明，旧药物cinanserin是SARS冠状病毒复制的抑制剂，最有可能通过抑制3CL蛋白酶发挥作用。
The 3C-like proteinase (3CL(pro)) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is one of the most promising targets for anti-SARS-CoV drugs due to its crucial role in the viral life cycle. In this study, a database containing structural information of more than 8,000 existing drugs was virtually screened by a docking approach to identify potential binding molecules of SARS-CoV 3CL(pro). As a target for screening, both a homology model and the crystallographic structure of the binding pocket of the enzyme were used. Cinanserin (SQ 10,643), a well-characterized serotonin antagonist that has undergone preliminary clinical testing in humans in the 1960s, showed a high score in the screening and was chosen for further experimental evaluation. Binding of both cinanserin and its hydrochloride to bacterially expressed 3CL(pro) of SARS-CoV and the related human coronavirus 229E (HCoV-229E) was demonstrated by surface plasmon resonance technology. The catalytic activity of both enzymes was inhibited with 50% inhibitory concentration (IC50) values of 5 mu M, as tested with a fluorogenic substrate. The antiviral activity of cinanserin was further evaluated in tissue culture assays, namely, a replicon system based on HCoV-229E and quantitative test assays with infectious SARS-CoV and HCoV-229E. All assays revealed a strong inhibition of coronavirus replication at nontoxic drug concentrations. The level of virus RNA and infectious particles was reduced by up to 4 log units, with IC50 values ranging from 19 to 34 mu M. These findings demonstrate that the old drug cinanserin is an inhibitor of SARS-CoV replication, acting most likely via inhibition of the 3CL proteinase.