Abstract：
The family of human-infecting coronaviruses (HCoVs) poses a serious threat to global health and includes several highly pathogenic strains that cause severe respiratory illnesses. It is essential that we develop effective broad-spectrum anti-HCoV agents to prepare for future outbreaks. In this study, we used PROteolysis TArgeting Chimera (PROTAC) technology focused on degradation of the HCoV main protease (Mpro), a conserved enzyme essential for viral replication and pathogenicity. By adapting the Mpro inhibitor GC376, we produced two novel PROTACs, P2 and P3, which showed relatively broad-spectrum activity against the human-infecting CoVs HCoV-229E, HCoV-OC43, and SARS-CoV-2. The concentrations of these PROTACs that reduced virus replication by 50 % ranged from 0.71 to 4.6 μM, and neither showed cytotoxicity at 100 μM. Furthermore, mechanistic binding studies demonstrated that P2 and P3 effectively targeted HCoV-229E, HCoV-OC43, and SARS-CoV-2 by degrading Mpro within cells in vitro. This study highlights the potential of PROTAC technology in the development of broad-spectrum anti-HCoVs agents, presenting a novel approach for dealing with future viral outbreaks, particularly those stemming from CoVs.
摘要：
人类感染冠状病毒（HCoV）家族对全球健康构成严重威胁，包括几种导致严重呼吸道疾病的高致病性菌株。我们必须开发有效的广谱抗HCoV药物，为未来的疫情做好准备。在这项研究中,我们使用ProteesolutionTrogeting嵌合体（PROTAC）技术专注于HCoV主要蛋白酶（Mpro）的降解，一种对病毒复制和致病性至关重要的保守酶。通过调整Mpro抑制剂GC376，我们生产了两种新型PROTACs，P2和P3对感染人类的CoVHCoV-229E显示出相对广谱的活性，HCoV-OC43和SARS-CoV-2。这些使病毒复制减少50％的PROTACs的浓度范围为0.71至4.6μM，并且在100μM时均未显示细胞毒性。此外，机制结合研究表明，P2和P3有效地靶向HCoV-229E，HCoV-OC43和SARS-CoV-2通过体外降解细胞内的Mpro。这项研究强调了PROTAC技术在开发广谱抗HCoV药物方面的潜力，提出了一种应对未来病毒爆发的新方法，特别是那些源于CoV的。
