Selective fluorescent nonpeptidic antagonists for vasopressin V(2) GPCR: application to ligand screening and oligomerization assays.
Fiche publication
Date publication
octobre 2012
Auteurs
Membres identifiés du Cancéropôle Est :
Pr HIBERT Marcel
Tous les auteurs :
Loison S, Cottet M, Orcel H, Adihou H, Rahmeh R, Lamarque L, Trinquet E, Kellenberger E, Hibert M, Durroux T, Mouillac B, Bonnet D
Lien Pubmed
Résumé
A series of fluorescent benzazepine ligands for the arginine-vasopressin V(2) receptor (AVP V(2)R) was synthesized using "Click" chemistry. Their in vitro pharmacological profile at AVP V(2)R, V(1a)R, V(1b)R, and oxytocin receptor was measured by binding assay and functional studies. Compound 9p, labeled with Lissamine Rhodamine B using novel solid-phase organic tagging (SPOrT) resin, exhibited a high affinity for V(2)R (4.0 nM), an excellent selectivity toward V(2)R and antagonist properties. By changing the nature of the dye, DY647 and Lumi4-Tb probes 44 and 47 still display a high affinity for V(2)R (5.6 and 5.8 nM, respectively). These antagonists constitute the first high-affinity selective nonpeptidic fluorescent ligands for V(2)R. They enabled the development of V(2)R time-resolved FRET-based assay readily amenable to high-throughput screening. Taking advantage of their selectivity, these compounds were also successfully involved in the study of V(1a)R-V(2)R dimerization on cell surface.
Référence
J Med Chem. 2012 Oct 25;55(20):8588-602