Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205, United States.
Department of Chemistry, Krieger School of Arts and Sciences, Johns Hopkins University, Baltimore, Maryland 21218, United States.
ACS Chem Biol. 2022 Jan 21;17(1):17-23. doi: 10.1021/acschembio.1c00721. Epub 2021 Dec 14.
Macrodomains are a class of conserved ADP-ribosylhydrolases expressed by viruses of pandemic concern, including coronaviruses and alphaviruses. Viral macrodomains are critical for replication and virus-induced pathogenesis; therefore, these enzymes are a promising target for antiviral therapy. However, no potent or selective viral macrodomain inhibitors currently exist, in part due to the lack of a high-throughput assay for this class of enzymes. Here we developed a high-throughput ADP-ribosylhydrolase assay using the SARS-CoV-2 macrodomain Mac1. We performed a pilot screen that identified dasatinib and dihydralazine as ADP-ribosylhydrolase inhibitors. Importantly, dasatinib inhibits SARS-CoV-2 and MERS-CoV Mac1 but not the closest human homologue, MacroD2. Our study demonstrates the feasibility of identifying selective inhibitors based on ADP-ribosylhydrolase activity, paving the way for the screening of large compound libraries to identify improved macrodomain inhibitors and to explore their potential as antiviral therapies for SARS-CoV-2 and future viral threats.
巨域结构域是一类保守的 ADP-核糖基水解酶，由具有大流行潜力的病毒表达，包括冠状病毒和甲病毒。病毒巨域结构域对于复制和病毒诱导的发病机制至关重要；因此，这些酶是抗病毒治疗的有希望的靶点。然而，目前没有有效的或选择性的病毒巨域抑制剂，部分原因是缺乏针对这类酶的高通量测定法。在这里，我们使用 SARS-CoV-2 的巨域 Mac1 开发了一种高通量 ADP-核糖基水解酶测定法。我们进行了一项初步筛选，发现达沙替尼和二氢氯嗪是 ADP-核糖基水解酶抑制剂。重要的是，达沙替尼抑制 SARS-CoV-2 和 MERS-CoV Mac1，但不抑制最接近的人类同源物 MacroD2。我们的研究证明了基于 ADP-核糖基水解酶活性鉴定选择性抑制剂的可行性，为筛选大型化合物库以鉴定改进的巨域抑制剂并探索它们作为 SARS-CoV-2 和未来病毒威胁的抗病毒治疗方法铺平了道路。
