Department of Pharmacology and Toxicology, College of Pharmacy, The University of Arizona, 1703 E Mabel St, Tucson, AZ, 85721-0207, USA.
Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, TX, 77555, USA.
Antiviral Res. 2022 Sep;205:105381. doi: 10.1016/j.antiviral.2022.105381. Epub 2022 Jul 11.
SARS-CoV-2 has raised the alarm to search for effective therapy for this virus. To date several vaccines have been approved but few available drugs reported recently still need approval from FDA. Remdesivir was approved for emergency use only. In this report, the SARS-CoV-2 3CLpro was expressed and purified. By using a FRET-based enzymatic assay, we have screened a library consisting of more than 300 different niclosamide derivatives and identified three molecules JMX0286, JMX0301, and JMX0941 as potent allosteric inhibitors against SARS-CoV-2 3CLpro, with IC values similar to that of known covalent inhibitor boceprevir. In a cell-based antiviral assay, these inhibitors can inhibit the virus growth with EC in the range of 2-3 μM. The mechanism of action of JMX0286, JMX0301, and JMX0941 were characterized by enzyme kinetics, affinity binding and protein-based substrate digestion. Molecular docking, molecular dynamics (MD) simulations and hydration studies suggested that JMX0286, JMX0301, JMX0941 bind specifically to an allosteric pocket of the SARS-CoV-2 3CL protease. This study provides three potent compounds for further studies.
SARS-CoV-2 引发了寻找针对该病毒有效疗法的警报。迄今为止，已有几种疫苗获得批准，但最近报道的少数可用药物仍需 FDA 批准。瑞德西韦仅获准紧急使用。在本报告中，我们表达和纯化了 SARS-CoV-2 3CLpro。通过使用基于 FRET 的酶促测定法，我们筛选了一个由超过 300 种不同尼氯柳胺衍生物组成的文库，并鉴定出三种分子 JMX0286、JMX0301 和 JMX0941 作为针对 SARS-CoV-2 3CLpro 的有效变构抑制剂，其 IC 值与已知共价抑制剂博赛泼维相似。在基于细胞的抗病毒测定中，这些抑制剂可以以 2-3 μM 的范围抑制病毒生长，EC 值。JMX0286、JMX0301 和 JMX0941 的作用机制通过酶动力学、亲和力结合和基于蛋白质的底物消化进行了表征。分子对接、分子动力学 (MD) 模拟和水合研究表明，JMX0286、JMX0301 和 JMX0941 特异性结合到 SARS-CoV-2 3CL 蛋白酶的变构口袋中。这项研究为进一步研究提供了三种有效化合物。
