蛋白酶是一种水解蛋白质和肽键的酶，因此它们控制几乎所有的生物过程。我们对蛋白酶功能的理解已经从非选择性的消化酶发展到高度专业化的分子剪刀，通过有限水解调控复杂的信号网络。蛋白酶的催化活性在多个层面上被严密地调节，从基因表达、运输和成熟到翻译后修饰。但是，当这种微妙的平衡被打破时，许多疾病就会产生，包括癌症、炎症性疾病、糖尿病和神经退行性疾病。对蛋白酶在病理生理学中的作用的新认识表明，这些酶代表了开发治疗性抑制剂和化学探针以可视化其冗余活性的优秀的分子靶点。最近，已经开发了许多平台技术来鉴定和优化蛋白酶底物和抑制剂，并以此为基础进一步开发化学探针和治疗药物。由于取得了巨大的成功，蛋白酶在治疗和诊断方面的临床潜力正在迅速增长，而且远未全部探索。因此，可以选择性地靶向异常的蛋白酶活性的小分子正在疾病细胞中出现。在本综述中，我们描述了蛋白酶药物设计以及小分子活性基探针的现代趋势，以在临床环境中可视化选择的蛋白酶。 版权所有©2022作者。由安徽省教育厅资助。 上海中医药大学.

Proteases are enzymes that hydrolyze peptide bonds in proteins and peptides; thus, they control virtually all biological processes. Our understanding of protease function has advanced considerably from nonselective digestive enzymes to highly specialized molecular scissors that orchestrate complex signaling networks through a limited proteolysis. The catalytic activity of proteases is tightly regulated at several levels, ranging from gene expression through trafficking and maturation to posttranslational modifications. However, when this delicate balance is disturbed, many diseases develop, including cancer, inflammatory disorders, diabetes, and neurodegenerative diseases. This new understanding of the role of proteases in pathologic physiology indicates that these enzymes represent excellent molecular targets for the development of therapeutic inhibitors, as well as for the design of chemical probes to visualize their redundant activity. Recently, numerous platform technologies have been developed to identify and optimize protease substrates and inhibitors, which were further used as lead structures for the development of chemical probes and therapeutic drugs. Due to this considerable success, the clinical potential of proteases in therapeutics and diagnostics is rapidly growing and is still not completely explored. Therefore, small molecules that can selectively target aberrant protease activity are emerging in diseases cells. In this review, we describe modern trends in the design of protease drugs as well as small molecule activity-based probes to visualize selected proteases in clinical settings.Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.