Wang Ping, Xia Yang-Liu, Zou Li-Wei, Qian Xing-Kai, Dou Tong-Yi, Jin Qiang, Li Shi-Yang, Yu Yang, Wang Dan-Dan, Luo Qun, Ge Guang-Bo, Yang Ling
Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, P. R. China.
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, P. R. China.
Chemistry. 2017 Aug 10;23(45):10800-10807. doi: 10.1002/chem.201701384. Epub 2017 Jun 27.
A practical two-photon fluorescent probe was developed for highly sensitive and selective sensing of the activities of catechol-O-methyltransferase (COMT) in complex biological samples. To this end, a series of 3-substituted 7,8-dihydroxycoumarins were designed and synthesized. Among them, 3-BTD displayed the best combination of selectivity, sensitivity, reactivity, and fluorescence response following COMT-catalyzed 8-O-methylation. The newly developed two-photon fluorescent probe 3-BTD can be used for determining the activities of COMT in complex biological samples and bio-imaging of endogenous COMT in living cells and tissue slices with good cell permeability, low cytotoxicity, and high imaging resolution. All these findings suggest that 3-BTD holds great promise for developing therapeutic molecules that target COMT, as well as for exploring COMT-associated biological processes and its biological functions in living systems. Furthermore, the strategy also sheds new light on the development of fluorescent probes for other conjugative enzymes.
开发了一种实用的双光子荧光探针，用于在复杂生物样品中高灵敏度和高选择性地检测儿茶酚-O-甲基转移酶（COMT）的活性。为此，设计并合成了一系列3-取代的7,8-二羟基香豆素。其中，3-BTD在COMT催化的8-O-甲基化反应后，展现出选择性、灵敏度、反应活性和荧光响应的最佳组合。新开发的双光子荧光探针3-BTD可用于测定复杂生物样品中COMT的活性，并对活细胞和组织切片中的内源性COMT进行生物成像，具有良好的细胞通透性、低细胞毒性和高成像分辨率。所有这些发现表明，3-BTD在开发靶向COMT的治疗分子以及探索COMT相关的生物过程及其在生命系统中的生物学功能方面具有巨大潜力。此外，该策略也为其他共轭酶荧光探针的开发提供了新的思路。
