Li Dongsheng, You Baoqing, Guo Keyu, Zhou Wenwen, Li Yan, Wang Chenyin, Chen Xiaofang, Wang Zhen, Zhang Jing, Si Shuyi
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Research Team of Molecular Medicine, The First Clinical Medical School of Shanxi Medical University, Taiyuan 030001, China.
Int J Mol Sci. 2025 Jan 15;26(2):678. doi: 10.3390/ijms26020678.
The recent coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has exerted considerable impact on global health. To prepare for rapidly mutating viruses and for the forthcoming pandemic, effective therapies targeting the critical stages of the viral life cycle need to be developed. Viruses are dependent on the interaction between the receptor-binding domain (RBD) of the viral Spike (S) protein (S-RBD) and the angiotensin-converting enzyme 2 (ACE2) receptor to efficiently establish infection and the following replicate. Targeting this interaction provides a promising strategy to inhibit the entry process of the virus, which in turn has both preventive and therapeutic effects. In this study, we developed a robust and straightforward assay based on the Yeast-Two Hybrid system (Y2H) for identifying inhibitors targeting the S-RBD-ACE2 interaction of SARS-CoV-2. Through high-throughput screening, two compounds were identified as potential entry inhibitors. Among them, IMB-1C was superior in terms of pseudovirus entry inhibition and toxicity. It could bind to both ACE2 and S-RBD and induce conformational change in the S-RBD+ACE2 complex. This is the first study to verify the feasibility of utilizing the Y2H system to discover potent SARS-CoV-2 inhibitors targeting the receptor recognition stage. This approach may also be applied in the discovery of other virus receptor recognition inhibitors.
由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019年新型冠状病毒病（COVID-19）大流行，已对全球健康产生了重大影响。为了应对快速变异的病毒以及即将到来的大流行，需要开发针对病毒生命周期关键阶段的有效疗法。病毒依赖于病毒刺突（S）蛋白的受体结合域（RBD）（S-RBD）与血管紧张素转换酶2（ACE2）受体之间的相互作用，以有效地建立感染并随后进行复制。针对这种相互作用提供了一种有前景的策略来抑制病毒的进入过程，这反过来具有预防和治疗作用。在本研究中，我们基于酵母双杂交系统（Y2H）开发了一种强大且直接的检测方法，用于鉴定针对SARS-CoV-2的S-RBD-ACE2相互作用的抑制剂。通过高通量筛选，鉴定出两种化合物作为潜在的进入抑制剂。其中，IMB-1C在假病毒进入抑制和毒性方面表现更优。它可以与ACE2和S-RBD结合，并诱导S-RBD+ACE2复合物的构象变化。这是第一项验证利用Y2H系统发现针对受体识别阶段的强效SARS-CoV-2抑制剂可行性的研究。这种方法也可能应用于发现其他病毒受体识别抑制剂。
