title: Selective fluorescent nonpeptidic antagonists for vasopressin V₂ GPCR: application to ligand screening and oligomerization assays.
abstract: A series of fluorescent benzazepine ligands for the arginine-vasopressin V₂ receptor (AVP V₂R) was synthesized using "Click" chemistry. Their in vitro pharmacological profile at AVP V₂R, 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 V₂R (4.0 nM), an excellent selectivity toward V₂R and antagonist properties. By changing the nature of the dye, DY647 and Lumi4-Tb probes 44 and 47 still display a high affinity for V₂R (5.6 and 5.8 nM, respectively). These antagonists constitute the first high-affinity selective nonpeptidic fluorescent ligands for V₂R. They enabled the development of V₂R 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-V₂R dimerization on cell surface.
authors.affiliation: Laboratoire d'Innovation Thérapeutique, UMR7200 CNRS/Université de Strasbourg, Faculté de Pharmacie, 74 Route du Rhin, 67412 Illkirch, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Loison
authors.forename: Martin
authors.initials: M
authors.lastname: Cottet
authors.forename: Hélène
authors.initials: H
authors.lastname: Orcel
authors.forename: Hélène
authors.initials: H
authors.lastname: Adihou
authors.forename: Rita
authors.initials: R
authors.lastname: Rahmeh
authors.forename: Laurent
authors.initials: L
authors.lastname: Lamarque
authors.forename: Eric
authors.initials: E
authors.lastname: Trinquet
authors.forename: Esther
authors.initials: E
authors.lastname: Kellenberger
authors.forename: Marcel
authors.initials: M
authors.lastname: Hibert
authors.forename: Thierry
authors.initials: T
authors.lastname: Durroux
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.forename: Dominique
authors.initials: D
authors.lastname: Bonnet
mesh_terms: D065092:Antidiuretic Hormone Receptor Antagonists; D001552:Benzazepines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D057930:Click Chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D008024:Ligands; D008958:Models, Molecular; D055503:Protein Multimerization; D018045:Receptors, Oxytocin / Q000037:antagonists & inhibitors / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000378:metabolism*; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: D065092:Antidiuretic Hormone Receptor Antagonists; D001552:Benzazepines; D005456:Fluorescent Dyes; D008024:Ligands; D018045:Receptors, Oxytocin; D017483:Receptors, Vasopressin