Human noroviruses (HuNoVs) are the most common cause of viral gastroenteritis resulting annually in ~219,000 deaths and a societal cost of ~USD 60 billion, and no antivirals or vaccines are available. Here, we assess the anti-norovirus activity of new peptidomimetic aldehydes related to the protease inhibitor rupintrivir. The early hit compound 4 inhibited the replication of murine norovirus (MNV) and the HuNoV GI.1 replicon in vitro (EC₅₀ ~ 1 µM) and swiftly cleared the HuNoV GI.1 replicon from the cells. Compound 4 still inhibits the proteolytic activity. We selected a resistant GI.1 replicon, with a mutation (I109V) in a highly conserved region of the viral protease, conferring a low yield of resistance against compound 4 and rupintrivir. After testing new derivatives, compound 10d was the most potent (EC₅₀ nanomolar range). Molecular docking indicated that the aldehyde group of compounds 4 and 10d bind with Cys139 in the HuNoV 3CL protease by a covalent linkage. Finally, compound 10d inhibited the replication of HuNoV GII.4 in infected zebrafish larvae, and PK studies in mice showed an adequate profile.

中文翻译：

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一类针对病毒蛋白酶的新型诺如病毒抑制剂，具有有效的体外和体内抗病毒活性

人类诺如病毒 (HuNoVs) 是病毒性肠胃炎的最常见原因，每年导致约 219,000 人死亡，社会成本约 600 亿美元，并且没有可用的抗病毒药物或疫苗。在这里，我们评估了与蛋白酶抑制剂 rupintrivir 相关的新型肽模拟醛的抗诺如病毒活性。早期的化合物4在体外抑制了鼠诺如病毒 (MNV) 和 HuNoV GI.1 复制子的复制 (EC ₅₀ ~ 1 µM)，并迅速从细胞中清除了 HuNoV GI.1 复制子。化合物4仍然抑制蛋白水解活性。我们选择了一种抗性 GI.1 复制子，在病毒蛋白酶的高度保守区域中具有突变 (I109V)，从而导致对化合物4的抗性产量低和鲁宾替韦。在测试新衍生物后，化合物10d是最有效的（EC ₅₀纳摩尔范围）。分子对接表明化合物4和10d的醛基通过共价键与HuNoV 3CL蛋白酶中的Cys139结合。最后，化合物10d在受感染的斑马鱼幼虫中抑制了 HuNoV GII.4 的复制，并且在小鼠中的 PK 研究显示了足够的概况。