Designing a Small Fluorescent Inhibitor to Investigate Soluble Epoxide Hydrolase Engagement in Living Cells.

Autor: Brunst S; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Schönfeld J; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Breunig P; Buchmann Institute for Molecular Life Sciences and Institute for Cell Biology and Neuroscience, Goethe University Frankfurt, Max-von-Laue-Strasse 15, 60438 Frankfurt, Germany., Burgers LD; Institute of Pharmaceutical Biology, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., DeMeglio M; Buchmann Institute for Molecular Life Sciences and Institute for Cell Biology and Neuroscience, Goethe University Frankfurt, Max-von-Laue-Strasse 15, 60438 Frankfurt, Germany., Ehrler JHM; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Lillich FF; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Weizel L; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Hefendehl JK; Buchmann Institute for Molecular Life Sciences and Institute for Cell Biology and Neuroscience, Goethe University Frankfurt, Max-von-Laue-Strasse 15, 60438 Frankfurt, Germany., Fürst R; Institute of Pharmaceutical Biology, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Proschak E; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany., Hiesinger K; Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Max-von-Laue-Strasse 9, 60438 Frankfurt, Germany.
Jazyk: angličtina
Zdroj: ACS medicinal chemistry letters [ACS Med Chem Lett] 2022 Jun 14; Vol. 13 (7), pp. 1062-1067. Date of Electronic Publication: 2022 Jun 14 (Print Publication: 2022).
DOI: 10.1021/acsmedchemlett.2c00073
Abstrakt: 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 50 < 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.
Competing Interests: The authors declare no competing financial interest.
(© 2022 The Authors. Published by American Chemical Society.)
Databáze: MEDLINE
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