Development and characterization of a fluorescent tracer for the free fatty acid receptor 2 (FFA2/GPR43)

Hansen, A. H., Sergeev, E. , Pandey, S. K., Hudson, B. D. , Christiansen, E., Milligan, G. and Ulven, T. (2017) Development and characterization of a fluorescent tracer for the free fatty acid receptor 2 (FFA2/GPR43). Journal of Medicinal Chemistry, 60(13), pp. 5638-5645. (doi: 10.1021/acs.jmedchem.7b00338) (PMID:28570808)

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The free fatty acid receptor 2 (FFA2/GPR43) is considered a potential target for treatment of metabolic and inflammatory diseases. Here we describe the development of the first fluorescent tracer for FFA2 intended as a tool for assessment of thermodynamic and kinetic binding parameters of unlabeled ligands. Starting with a known azetidine FFA2 antagonist, we used a carboxylic acid moiety known not to be critical for receptor interaction as attachment point for a nitrobenzoxadiazole (NBD) fluorophore. This led to the development of 4 (TUG-1609), a fluorescent tracer for FFA2 with favorable spectroscopic properties and high affinity, as determined by bioluminescence resonance energy transfer (BRET)-based saturation and kinetic binding experiments, as well as a high specific to nonspecific BRET binding signal. A BRET-based competition binding assay with 4 was also established and used to determine binding constants and kinetics of unlabeled ligands.

Item Type:Articles
Additional Information:The study has been funded by the Danish Council for Strategic Research (grant 11-116196) and the University of Southern Denmark. EC is supported by a fellowship from the Lundbeck Foundation. BDH is supported by a fellowship from the University of Glasgow.
Keywords:Molecular medicine, drug discovery.
Glasgow Author(s) Enlighten ID:Hudson, Dr Brian and Sergeev, Miss Eugenia and Milligan, Professor Graeme
Authors: Hansen, A. H., Sergeev, E., Pandey, S. K., Hudson, B. D., Christiansen, E., Milligan, G., and Ulven, T.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Journal of Medicinal Chemistry
Publisher:American Chemical Society (ACS)
ISSN (Online):1520-4804
Published Online:01 June 2017
Copyright Holders:Copyright © 2017 American Chemical Society
First Published:First published in Journal of Medicinal Chemistry 60(13):5638-5645
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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