Identification of a tetrahydroquinoline analog as a pharmacological inhibitor of the cAMP-binding protein Epac.

Delphine Courilleau 1 Malik Bisserier 2 Jean-Christophe Jullian 3 Alexandre Lucas 2 Pascal Bouyssou 4 Rodolphe Fischmeister 1 Jean-Paul Blondeau 1 Frank Lezoualc'H 2, *
* Auteur correspondant
1 Signalisation et physiopathologie cardiaque
2 I2MC - Institut des Maladies Métaboliques et Cardiovasculaires
3 MBCIS - Molécules bioactives, conception, isolement et synthèse
4 ICOA - Institut de Chimie Organique et Analytique
Abstract : The cAMP-binding protein Epac is a therapeutic target for the treatment of various diseases such as cardiac hypertrophy and tumor invasion. This points out the importance to develop Epac inhibitors to better understand the involvement of these cAMP sensors in physiology and pathophysiology. Here, we have developed a functional fluorescence-based high-throughput assay with a Z' value around 0.7 for screening Epac-specific antagonists. We identified an Epac1 inhibitor compound named CE3F4 that blocked Epac1 guanine nucleotide exchange activity toward its effector Rap1 both in cell-free systems and in intact cells. CE3F4 is a tetrahydroquinoline analog that fails to influence protein kinase A holoenzyme activity. CE3F4 inhibited neither the interaction of Rap1 with Epac1 nor directly the GDP exchange on Rap1. The kinetics of inhibition by CE3F4 indicated that this compound did not compete for binding of agonists to Epac1 and suggested an uncompetitive inhibition mechanism with respect to Epac1 agonists. A structure-activity study showed that the formyl group on position 1 and the bromine atom on position 5 of the tetrahydroquinoline skeleton were important for CE3F4 to exert its inhibitory activity. Finally, CE3F4 inhibited Rap1 activation in living cultured cells, following Epac activation by either 8-(4-chlorophenylthio)-2'-O-methyl-cAMP, an Epac-selective agonist, or isoprenaline, a non-selective β-adrenergic receptor agonist. Our study shows that CE3F4 and related compounds may serve as a basis for the development of new therapeutic drugs.
Keywords : Cyclic AMP (cAMP) G Protein-coupled Receptors (GPCR) Guanine Nucleotide Exchange Factor (GEF) Molecular Pharmacology Small GTPases CE3F4 Epac
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Article dans une revue
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (53), pp.44192-202. 〈10.1074/jbc.M112.422956〉
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Contributeur : Marie Francoise Simon <>
Soumis le : jeudi 19 juin 2014 - 14:54:43
Dernière modification le : jeudi 13 septembre 2018 - 10:50:02

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Delphine Courilleau, Malik Bisserier, Jean-Christophe Jullian, Alexandre Lucas, Pascal Bouyssou, et al.. Identification of a tetrahydroquinoline analog as a pharmacological inhibitor of the cAMP-binding protein Epac.. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (53), pp.44192-202. 〈10.1074/jbc.M112.422956〉. 〈inserm-01010289〉

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