Structural Basis for Antiviral Inhibition of the Main Protease, 3C, from Human Enterovirus 93
Structural Basis for Antiviral Inhibition of the Main Protease, 3C, from Human Enterovirus 93
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DOI：
10.1128/jvi.05062-11
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发表时间：
2011-10-01
影响因子：
5.4
通讯作者：
Coll, Miquel
中科院分区：
文献类型：
--
作者：
Costenaro, Lionel;Kaczmarska, Zuzanna;Coll, Miquel
小核糖核酸病毒科肠道病毒属的成员非常丰富，常见的人类病原体属于鼻病毒（HRV）和肠道病毒（EV）种，包括不同的埃可病毒、柯萨奇病毒和脊髓灰质炎病毒。它们引起广泛的临床表现，从无症状到具有神经和/或心脏表现的严重疾病。EV的大流行爆发可能伴有脑膜炎和/或瘫痪，并且可能是致命的。然而，没有针对大多数EV的有效预防或抗病毒治疗。EV RNA基因组指导单个多聚蛋白的合成，所述多聚蛋白通过3C蛋白酶（3C（pro））在Gln下箭头Gly切割位点处自催化加工成成熟蛋白，所述3C蛋白酶具有窄的保守底物特异性。这些裂解对于病毒复制是必不可少的，使得3C（pro）成为抗病毒药物开发的极好靶点。在这项研究中，我们报告的第一个确定的晶体结构的3C（亲）从肠道病毒B，EV-93，最近确定的病原体，单独和在复杂的抗HRV分子化合物1（AG 7404）和鲁帕韦（AG 7088）的分辨率分别为1.9，1.3和1.5埃。EV-93 3C（pro）采用胰凝乳蛋白酶样折叠，具有与犀牛病毒、柯萨奇病毒和脊髓灰质炎病毒3C蛋白酶相似的正离子构型的氧阴离子孔和底物结合口袋。我们表明化合物1和鲁帕韦在感染细胞中均具有抗EV-93的活性，并在体外抑制EV-93 3C（pro）的蛋白水解活性。这些结果提供了一个框架，进一步结构导向优化的测试化合物，以产生抗病毒药物对广泛的EV物种。
Members of the Enterovirus genus of the Picornaviridae family are abundant, with common human pathogens that belong to the rhinovirus (HRV) and enterovirus (EV) species, including diverse echo-, coxsackie-and polioviruses. They cause a wide spectrum of clinical manifestations ranging from asymptomatic to severe diseases with neurological and/or cardiac manifestations. Pandemic outbreaks of EVs may be accompanied by meningitis and/or paralysis and can be fatal. However, no effective prophylaxis or antiviral treatment against most EVs is available. The EV RNA genome directs the synthesis of a single polyprotein that is autocatalytically processed into mature proteins at Gln down arrow Gly cleavage sites by the 3C protease (3C(pro)), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3C(pro) an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3C(pro) from an enterovirus B, EV-93, a recently identified pathogen, alone and in complex with the anti-HRV molecules compound 1 (AG7404) and rupintrivir (AG7088) at resolutions of 1.9, 1.3, and 1.5 angstrom, respectively. The EV-93 3C(pro) adopts a chymotrypsin-like fold with a canonically configured oxyanion hole and a substrate binding pocket similar to that of rhino-, coxsackie-and poliovirus 3C proteases. We show that compound 1 and rupintrivir are both active against EV-93 in infected cells and inhibit the proteolytic activity of EV-93 3C(pro) in vitro. These results provide a framework for further structure-guided optimization of the tested compounds to produce antiviral drugs against a broad range of EV species.