上海科技大学杨海涛团队报道了猴痘蛋白酶、核心蛋白酶的底物识别和裂解机制。该研究于2025年4月22日发表于国际一流学术期刊《自然》杂志上。

该课题组人员猴痘病毒（MPXV）apo-CorePro的结构及其与抑制剂阿洛司他丁的复合物，阿洛司特丁是肌营养不良的候选药物2。这些结构表明，CorePro形成了一种具有独特“舞蹈夫妇”折叠的同型二聚体。CorePro的催化中间态由天然底物（I-G18）的醛衍生物表征。该衍生物与催化型半胱氨酸328 （Cys328）共价结合，使病毒蛋白酶的活性位点在底物结合时从载子形式的封闭构象转变为有利的开放构象。基于CorePro-I-G18配合物，研究小组设计了一系列具有腈弹头的拟肽抑制剂，这些抑制剂可以与催化Cys328共价锚定。这些化合物抑制CorePro的IC50值为44.9 ~100.3nM，具有较强的抗痘病毒活性。他们的研究为设计抗痘病毒感染的广谱抑制剂提供了基础。

据了解，痘病毒引起包括天花和猴痘在内的严重疾病，对人类健康构成重大威胁。痘病毒核心蛋白酶（CorePro）对病毒成熟至关重要，在痘病毒中高度保守，使其成为有吸引力的抗病毒靶点。然而，CorePro的结构仍然未知，阻碍了抗病毒药物的开发。

附：英文原文

Title: Substrate recognition and cleavage mechanism of the monkeypox protease, Core protease

Author: Gao, Yan, Xie, Xiong, Zhang, Xiaoyu, Cao, Junyuan, Lan, Weiqi, You, Tian, Li, Dongxu, Dong, Xuxue, Dai, Wenhao, Xiang, Yingchun, Hu, Shulei, Shang, Weijuan, Wu, Botao, Zhang, Yumin, Xu, Jin, Liu, Xiaoce, Wang, Haofeng, Hu, Wanlong, Zhang, Mingjing, Duan, Yinkai, Cui, Wen, Zhou, Hao, Mao, Shengjiang, Jia, Handi, Sun, Zhanqi, Jia, Menghan, Yin, Yue, Nguyen, Henry C., Yang, Kailin, Yang, Bei, Yang, Xiuna, Ji, Xiaoyun, Xiao, Gengfu, Wang, Wei, Zhang, Leike, Rao, Zihe, Liu, Hong, Yang, Haitao

Issue&Volume: 2025-04-22

Abstract: Poxviruses cause severe diseases including smallpox and mpox, which pose major threats to human health. The poxvirus core protease (CorePro) is essential for viral maturation and highly conserved in poxviruses, making it an attractive antiviral target1. However, the structure of CorePro remains unknown, hampering antiviral development. Here, we determined the structures of mpox virus (MPXV) apo-CorePro and its complex with an inhibitor aloxistatin, which is a drug candidate for muscular dystrophy2. These structures show that CorePro forms a homodimer featuring a unique “dancing-couple” fold. The catalytic intermediate state of CorePro was characterized by an aldehyde derivative from a natural substrate (I-G18). This derivative covalently binds to the catalytic cysteine 328 (Cys328), making the active site of viral protease shift from a closed conformation in the apo-form to a favorable open conformation upon substrate binding. Based on the CorePro-I-G18 complex, we then designed a series of peptidomimetic inhibitors with a nitrile warhead, which could covalently anchor with the catalytic Cys328. These compounds inhibit the CorePro with IC50 values of 44.9-100.3 nM, exhibiting potent and broad anti-poxvirus activity as well. Our studies provide the basis for designing wide-spectrum inhibitors against poxvirus infections.

DOI: 10.1038/s41586-025-09014-x

Source: https://www.nature.com/articles/s41586-025-09014-x