PDF(Pubmed)
Abstract：
The emergence of severe acute respiratory syndrome coronavirus 2, the causative agent of coronavirus disease 2019, has resulted in the largest pandemic in recent history. Current therapeutic strategies to mitigate this disease have focused on the development of vaccines and on drugs that inhibit the viral 3CL protease or RNA-dependent RNA polymerase enzymes. A less-explored and potentially complementary drug target is Nsp15, a uracil-specific RNA endonuclease that shields coronaviruses and other nidoviruses from mammalian innate immune defenses. Here, we perform a high-throughput screen of over 100,000 small molecules to identify Nsp15 inhibitors. We characterize the potency, mechanism, selectivity, and predicted binding mode of five lead compounds. We show that one of these, IPA-3, is an irreversible inhibitor that might act via covalent modification of Cys residues within Nsp15. Moreover, we demonstrate that three of these inhibitors (hexachlorophene, IPA-3, and CID5675221) block severe acute respiratory syndrome coronavirus 2 replication in cells at subtoxic doses. This study provides a pipeline for the identification of Nsp15 inhibitors and pinpoints lead compounds for further development against coronavirus disease 2019 and related coronavirus infections.
摘要：
SARS-CoV-2的出现，是COVID-19的病原体，导致了近代史上最大的大流行。目前缓解这种疾病的治疗策略集中在疫苗和抑制病毒3CL蛋白酶或RNA依赖性RNA聚合酶的药物的开发上。一个较少探索和潜在的补充药物靶标是Nsp15，这是一种尿嘧啶特异性RNA内切核酸酶，可保护冠状病毒和其他nido病毒免受哺乳动物先天免疫防御。这里,我们对超过100,000个小分子进行高通量筛选以鉴定Nsp15抑制剂。我们表征了效力，机制，选择性,并预测了五种先导化合物的结合模式。我们展示了其中一个，IPA-3是一种不可逆的抑制剂，可能通过共价修饰Nsp15中的Cys残基起作用。此外，我们证明了其中三种抑制剂(六氯酚，IPA-3和CID5675221）在亚毒性剂量下阻止SARS-CoV-2在细胞中的复制。本研究为Nsp15抑制剂的鉴定提供了管道，并确定了用于进一步开发抗COVID-19和相关冠状病毒感染的先导化合物。
