Brunst Steffen, Schönfeld Julia, Breunig Peter, Burgers Luisa D, DeMeglio Murphy, Ehrler Johanna H M, Lillich Felix F, Weizel Lilia, Hefendehl Jasmin K, Fürst Robert, Proschak Ewgenij, Hiesinger Kerstin
Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany.
Buchmann Institute for Molecular Life Sciences and Institute for Cell Biology and Neuroscience, Goethe University Frankfurt, Max-von-Laue-Strasse 15, 60438 Frankfurt, Germany.
ACS Med Chem Lett. 2022 Jun 14;13(7):1062-1067. doi: 10.1021/acsmedchemlett.2c00073. eCollection 2022 Jul 14.
Soluble epoxide hydrolase (sEH) is a promising target for a number of inflammation-related diseases. In addition, inhibition of sEH has been shown to reduce neuroinflammation, which plays a critical role in the development of central nervous system (CNS) diseases such as Alzheimer's disease. In this study, we present the rational design of a small fluorescent sEH inhibitor. Starting from the clinical candidate GSK2256294A, we replaced the triazine moiety with the 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) fluorophore. The resulting fluorescent sEH inhibitor displayed excellent potency in an in vitro enzyme activity assay (IC < 2 nM). The developed inhibitor is applicable in a NanoBRET-based assay system suitable for studying sEH target engagement in living cells. Furthermore, the inhibitor can be used to visualize sEH in sEH-transfected HEK293 cells and in primary mouse astrocytes by fluorescence microscopy.
可溶性环氧化物水解酶（sEH）是多种炎症相关疾病的一个有前景的靶点。此外，已表明抑制sEH可减轻神经炎症，而神经炎症在诸如阿尔茨海默病等中枢神经系统（CNS）疾病的发展中起关键作用。在本研究中，我们展示了一种小型荧光sEH抑制剂的合理设计。从临床候选药物GSK2256294A出发，我们用4-氯-7-硝基苯并-2-恶唑-1,3-二唑（NBD-Cl）荧光团取代了三嗪部分。所得的荧光sEH抑制剂在体外酶活性测定中表现出优异的效力（IC<2 nM）。所开发的抑制剂适用于基于纳米生物发光共振能量转移（NanoBRET）的测定系统，该系统适用于研究活细胞中的sEH靶点结合情况。此外，该抑制剂可通过荧光显微镜用于在转染了sEH的HEK293细胞和原代小鼠星形胶质细胞中可视化sEH。
