Sanders Brian, Pokhrel Suman, Labbe Audrey, Mathews Irimpan, Cooper Connor, Davidson Russell, Phillips Gwyndalyn, Weiss Kevin, Zhang Qiu, O'Neill Hugh, Kaur Manat, Ferrins Lori, Schmidt Jurgen, Reichard Walter, Surendranathan Surekha, Parvathareddy Jyothi, Phillips Lexi, Rainville Christopher, Sterner David, Kumaran Desigan, Andi Babak, Babnigg Gyorgy, Moriarrty Nigel, Adams Paul, Joachimiak Andrzej, Hurst Brett, Kumar Suresh, Butt Tauseef, Jonsson Colleen, Wakatsuki Soichi, Galanie Stephanie, Head Martha, Parks Jerry
Oak Ridge National Laboratory.
Stanford University.
Res Sq. 2022 Jul 21:rs.3.rs-906621. doi: 10.21203/rs.3.rs-906621/v1.
Direct-acting antivirals are needed to combat coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). The papain-like protease (PLpro) domain of Nsp3 from SARS-CoV-2 is essential for viral replication. In addition, PLpro dysregulates the host immune response by cleaving ubiquitin and interferon-stimulated gene 15 protein (ISG15) from host proteins. As a result, PLpro is a promising target for inhibition by small-molecule therapeutics. Here we have designed a series of covalent inhibitors by introducing a peptidomimetic linker and reactive electrophile onto analogs of the noncovalent PLpro inhibitor GRL0617. The most potent compound inhibited PLpro with / = 10,000 M s, achieved sub-μM EC values against three SARS-CoV-2 variants in mammalian cell lines, and did not inhibit a panel of human deubiquitinases at > 30 μM concentrations of inhibitor. An X-ray co-crystal structure of the compound bound to PLpro validated our design strategy and established the molecular basis for covalent inhibition and selectivity against structurally similar human DUBs. These findings present an opportunity for further development of covalent PLpro inhibitors.
需要直接作用抗病毒药物来对抗由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）。SARS-CoV-2的Nsp3的木瓜蛋白酶样蛋白酶（PLpro）结构域对病毒复制至关重要。此外，PLpro通过从宿主蛋白上切割泛素和干扰素刺激基因15蛋白（ISG15）来失调宿主免疫反应。因此，PLpro是小分子治疗药物抑制的一个有前景的靶点。在这里，我们通过在非共价PLpro抑制剂GRL0617的类似物上引入拟肽连接子和反应性亲电试剂设计了一系列共价抑制剂。最有效的化合物以/ = 10,000 M s的速度抑制PLpro，在哺乳动物细胞系中对三种SARS-CoV-2变体的EC值达到亚微摩尔，并且在抑制剂浓度> 30 μM时不抑制一组人去泛素化酶。与PLpro结合的化合物的X射线共晶体结构验证了我们的设计策略，并建立了共价抑制和对结构相似的人DUBs选择性的分子基础。这些发现为共价PLpro抑制剂的进一步开发提供了机会。
