Development and Characterization of a Fluorescent Tracer for the Free Fatty Acid Receptor 2 (FFA2/GPR43)

Anders Højgaard Hansen; Eugenia Sergeev; Sunil K. Pandey; Brian D Hudson; Elisabeth Rexen Ulven; Graeme Milligan; Trond Ulven

doi:10.1021/acs.jmedchem.7b00338

Development and Characterization of a Fluorescent Tracer for the Free Fatty Acid Receptor 2 (FFA2/GPR43)

Anders Højgaard Hansen, Eugenia Sergeev, Sunil K. Pandey, Brian D Hudson, Elisabeth Rexen Ulven, Graeme Milligan, Trond Ulven

22 Citationer (Scopus)

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.

Originalsprog	Engelsk
Tidsskrift	Journal of Medicinal Chemistry
Vol/bind	60
Udgave nummer	13
Sider (fra-til)	5638-5645
Antal sider	8
ISSN	0022-2623
DOI	https://doi.org/10.1021/acs.jmedchem.7b00338
Status	Udgivet - 13 jul. 2017
Udgivet eksternt	Ja

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10.1021/acs.jmedchem.7b00338

Citationsformater

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title = "Development and Characterization of a Fluorescent Tracer for the Free Fatty Acid Receptor 2 (FFA2/GPR43)",

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.",

author = "Hansen, {Anders H{\o}jgaard} and Eugenia Sergeev and Pandey, {Sunil K.} and Hudson, {Brian D} and {Rexen Ulven}, Elisabeth and Graeme Milligan and Trond Ulven",

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AU - Hansen, Anders Højgaard

AU - Sergeev, Eugenia

AU - Pandey, Sunil K.

AU - Hudson, Brian D

AU - Rexen Ulven, Elisabeth

AU - Milligan, Graeme

AU - Ulven, Trond

PY - 2017/7/13

Y1 - 2017/7/13

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AB - 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.

U2 - 10.1021/acs.jmedchem.7b00338

DO - 10.1021/acs.jmedchem.7b00338

M3 - Journal article

C2 - 28570808

SN - 0022-2623

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SP - 5638

EP - 5645

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

IS - 13

ER -

Development and Characterization of a Fluorescent Tracer for the Free Fatty Acid Receptor 2 (FFA2/GPR43)

Abstract

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