α-Ketoamides as Broad-Spectrum Inhibitors of Coronavirus and Enterovirus Replication: Structure-Based Design, Synthesis, and Activity Assessment
α-Ketoamides as Broad-Spectrum Inhibitors of Coronavirus and Enterovirus Replication: Structure-Based Design, Synthesis, and Activity Assessment
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DOI：
10.1021/acs.jmedchem.9b01828
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发表时间：
2020-05-14
影响因子：
7.3
通讯作者：
Hilgenfeld, Rolf
中科院分区：
文献类型：
--
作者：
Zhang, Linlin;Lin, Daizong;Hilgenfeld, Rolf
冠状病毒的主要蛋白酶和肠道病毒的3C酶具有相似的活性部位结构，对底物P1位的谷氨酰胺有独特的要求。由于其独特的特异性和在病毒多蛋白加工中的重要作用，这些酶是抗病毒药物开发的合适靶点。为了获得几乎等效的、广谱的抗甲-冠状病毒、β-冠状病毒和肠道病毒的抗病毒药物，我们进行了一种基于结构的模拟多肽的α-酮酰胺类化合物作为主酶和3C酶的抑制剂。作为本研究的一部分，我们确定了六种蛋白酶-抑制剂复合体的晶体结构。合成的化合物被用来对抗重组酶，以及在病毒复制体和病毒感染的细胞培养中进行测试；大多数化合物对细胞没有毒性。事实证明，优化α-酮酰胺的P2取代基对于实现对这三个病毒属的近乎等效性至关重要。在细胞培养中，最好的近效抑制剂11u(P2=环戊基甲基)和11r(P2=环己基甲基)对肠道病毒、甲冠状病毒和β冠状病毒显示出低微摩尔的EC50值。在Huh7细胞中，11R对中东呼吸综合征冠状病毒表现出三位数的皮摩尔活性。
The main protease of coronaviruses and the 3C protease of enteroviruses share a similar active-site architecture and a unique requirement for glutamine in the P1 position of the substrate. Because of their unique specificity and essential role in viral polyprotein processing, these proteases are suitable targets for the development of antiviral drugs. In order to obtain near-equipotent, broad-spectrum antivirals against alphacoronaviruses, betacoronaviruses, and enteroviruses, we pursued a structure-based design of peptidomimetic alpha-ketoamides as inhibitors of main and 3C proteases. Six crystal structures of protease-inhibitor complexes were determined as part of this study. Compounds synthesized were tested against the recombinant proteases as well as in viral replicons and virus-infected cell cultures; most of them were not cell-toxic. Optimization of the P2 substituent of the alpha-ketoamides proved crucial for achieving near-equipotency against the three virus genera. The best near-equipotent inhibitors, 11u (P2 = cyclopentylmethyl) and 11r (P2 = cyclo-hexylmethyl), display low-micromolar EC50 values against enteroviruses, alphacoronaviruses, and betacoronaviruses in cell cultures. In Huh7 cells, 11r exhibits three-digit picomolar activity against the Middle East Respiratory Syndrome coronavirus.