The cytochrome P450 enzymes (P450s or CYPs) are a class of heme-containing monooxygenases responsible for the oxidative metabolism of a wide range of endogenous substances and foreign chemicals. Deciphering the physiological functions of CYPs and their relevance to human diseases requires reliable tools for sensing target enzyme in complex biological systems. Over the past few decades, great efforts have been made to develop isoform-specific probe substrates for human CYP(s) and use them in both basic researches and translational applications. The growing database of the ligands and 3D-structures of human CYPs strongly facilitates the design and development of CYP probe substrates with improved specificity. Herein, we summarize the recent progress in the development and applications of molecular probes for human CYPs. The structural and catalytic features of human CYPs, as well as the design principles and strategies used in the development of CYP probe substrates, have been well summarized. Two comprehensive lists of molecular probes for various human CYP enzymes, including non-optical and optical probe substrates, along with their structural information and kinetic parameters, are presented. Finally, the current challenges and future perspectives in this field are proposed and highlighted. The information and knowledge presented here provides practical tools for sensing CYP activities in complex biological systems, exploring the relevance of CYPs to human diseases, as well as screening and characterizing of CYP modulators.

细胞色素P450酶（P450或CYP）是一类含血红素的单加氧酶，负责多种内源性物质和外来化学物质的氧化代谢。解释CYP的生理功能及其与人类疾病的相关性，需要可靠的工具来感测复杂生物系统中的目标酶。在过去的几十年中，为开发用于人CYP的同工型特异性探针底物并在基础研究和翻译应用中使用它们做出了巨大的努力。人CYP配体和3D结构的数据库不断增长，极大地促进了CYP探针底物的设计和开发，并提高了特异性。本文中，我们总结了人类CYP分子探针的开发和应用的最新进展。已经很好地总结了人CYP的结构和催化特性，以及用于CYP探针底物开发的设计原理和策略。列出了两种针对各种人类CYP酶的分子探针的完整清单，包括非光学和光学探针底物，以及它们的结构信息和动力学参数。最后，提出并强调了该领域的当前挑战和未来前景。此处提供的信息和知识为检测复杂生物系统中CYP活性，探索CYP与人类疾病的相关性以及CYP调节剂的筛选和表征提供了实用的工具。已经很好地总结了。列出了两种针对各种人类CYP酶的分子探针的完整清单，包括非光学和光学探针底物，以及它们的结构信息和动力学参数。最后，提出并强调了该领域的当前挑战和未来前景。此处提供的信息和知识为检测复杂生物系统中CYP活性，探索CYP与人类疾病的相关性以及CYP调节剂的筛选和表征提供了实用的工具。已经很好地总结了。列出了两种针对各种人类CYP酶的分子探针的完整清单，包括非光学和光学探针底物，以及它们的结构信息和动力学参数。最后，提出并强调了该领域的当前挑战和未来前景。此处提供的信息和知识为检测复杂生物系统中CYP活性，探索CYP与人类疾病的相关性以及CYP调节剂的筛选和表征提供了实用的工具。提出并强调了该领域的当前挑战和未来前景。此处提供的信息和知识为检测复杂生物系统中CYP活性，探索CYP与人类疾病的相关性以及CYP调节剂的筛选和表征提供了实用的工具。提出并强调了该领域的当前挑战和未来前景。此处提供的信息和知识为检测复杂生物系统中CYP活性，探索CYP与人类疾病的相关性以及CYP调节剂的筛选和表征提供了实用的工具。