羧酸酯酶1(CES1)参与众多外源物和内源性脂类的代谢，并与多种代谢性疾病的发生发展密切相关。长期以来，业界一直缺少高效、灵敏且能实时动态监测复杂生命体系中CES1活性的检测方法。为此，本项目拟在前期工作基础上，从生物发光母体的改良和底物的多属性优化两方面入手，采用计算机辅助设计和实验手段相结合的策略，系统开展D-萤光素酯类候选底物的理性设计、结构改造和多属性优化等研究。旨在通过研究策略和方法学的创新，设计研发出属性优良且适于复杂生物体系的新型CES1生物发光底物。在此基础上，针对不同的研究对象和实验目的，构建一整套实用的CES1生化检测方法并将其用于CES1抑制剂/诱导剂的高通量筛选与评价、复杂生物体系中CES1活性的原位检测和可视化分析等研究。上述研究不仅有望为药理学、化学生物学及新药研发等领域提供实用的工具分子和生化检测方法，同时还可为设计研发其它丝氨酸水解酶的特异性光学底物提供借鉴和指导。

Carboxylesterase 1 (CES1) is a key enzyme responsible for the hydrolysis of a wide variety of xenobiotics and endogenous esters, and this enzyme is also closely related to the occurrence and development of various metabolic diseases. Until now, the highly efficient, sensitive, and real-time dynamic methods for sensing CES1 activity in complex biological systems are rarely reported. On the basis of our previous studies, this project aimed to design and develop a novel CES1 bioluminescent substrate with excellent properties, via modification of D-fluorescein as a basic scaffold and optimization of the multiple properties of candidate CES1 probe substrates. The rational design, structural modification and optimization of D-fluorescein esters as candidate CES1 probe substrates will be systematically investigated, with the help of computer-aided design and a panel of experimental techniques. Following innovating research strategies or methodologies, our studies may get a novel CES1 bioluminescent substrate with excellent properties, which will be used for sensing CES1 in complex biological systems. After that, a set of practical biochemical assays for CES1 will be constructed on the basis of different samples and purposes, such assays will be used for high-throughput screening and characterization of CES1 inhibitors or inducers, as well as in situ monitoring and imaging analysis of CES1 activity in complex biological systems. Taken together, these studies are expected to develop practical tools and biochemical detection methods for sensing CES1, which will be very helpful for CES1-associated studies in the fields of pharmacology, chemical biology and drug discovery, as well as provide experience and guidance for the design and development of specific optical substrates for other serine hydrolases.