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↓Gly cleavage sites by the 3C protease (3Cpro), which has narrow, conserved substrate specificity. These cleavages are essential for virus replication, making 3Cpro an excellent target for antivirus drug development. In this study, we report the first determination of the crystal structure of 3Cpro 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 Å, respectively. The EV-93 3Cpro 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 3Cpro 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.
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机译:Picornaviridae家族的肠病毒属成员很多,常见的人类病原体属于鼻病毒(HRV)和肠病毒(EV)物种,包括多种回声,柯萨奇和脊髓灰质炎病毒。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。电动车大流行爆发可能伴有脑膜炎和/或瘫痪,并可能致命。但是,没有针对大多数电动车的有效预防或抗病毒治疗。 EV RNA基因组指导单个多蛋白的合成,该蛋白通过3C蛋白酶(3C pro sup>)在Gln↓Gly裂解位点自动催化加工成成熟蛋白,该酶具有狭窄的保守底物特异性。这些裂解对于病毒复制至关重要,使3C pro sup>成为抗病毒药物开发的绝佳靶标。在这项研究中,我们报告了由肠道病毒B EV-93(一种最近鉴定出的病原体)单独和与抗HRV分子化合物1配合使用时首次确定3C pro sup>的晶体结构的方法( AG7404和rupintrivir(AG7088)的分辨率分别为1.9、1.3和1.5Å。 EV-93 3C pro sup>采用类似于胰凝乳蛋白酶的折叠,带有一个标准配置的氧阴离子孔和一个类似于犀牛,柯萨奇和脊髓灰质炎3C蛋白酶的底物结合袋。我们发现化合物1和rupintrivir都对被感染细胞中的EV-93具有活性,并在体外抑制EV-93 3C pro sup>的蛋白水解活性。这些结果为进一步测试化合物的结构指导优化提供了一个框架,以生产针对多种EV物种的抗病毒药物。
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