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)
|
Text
144377.pdf - Accepted Version 771kB |
Abstract
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. |
| Status: | Published |
| Refereed: | Yes |
| 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 > School of Molecular Biosciences |
| Journal Name: | Journal of Medicinal Chemistry |
| Publisher: | American Chemical Society (ACS) |
| ISSN: | 0022-2623 |
| 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 |
University Staff: Request a correction | Enlighten Editors: Update this record
Altmetric
Altmetric