title: Toward efficient drug screening by homogeneous assays based on the development of new fluorescent vasopressin and oxytocin receptor ligands.
abstract: A series of fluorescent ligands designed for vasopressin and oxytocin G protein-coupled receptors was synthesized and characterized to develop fluorescence polarization or homogeneous time-resolved fluorescence (HTRF) binding assays. These ligands, labeled with europium pyridine-bis-bipyridine cryptate or with Alexa 488,546,647 selectively bound to the vasopressin V1a and oxytocin receptors with high affinities and exhibited antagonistic properties. The affinities of several unlabeled ligands determined by our homogeneous assays on membrane preparations or on intact cells into 96- and 384-well plate formats were similar to those determined by usual radioligand binding methods. Compared to other binding assays, the polarization and HTRF binding assays are nonradiaoactive, therefore safer to perform, yet very sensitive and homogeneous, therefore easier and faster to automate. These methods are thus suitable for efficient drug high-throughput screening procedures and can easily be applied to other G protein-coupled receptor models.
authors.affiliation: Institut de Génomique Fonctionnelle, Montpellier, France, CNRS UMR5203, Montpellier, France, INSERM, U661, Montpellier, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Albizu
authors.forename: Géraldine
authors.initials: G
authors.lastname: Teppaz
authors.forename: René
authors.initials: R
authors.lastname: Seyer
authors.forename: Hervé
authors.initials: H
authors.lastname: Bazin
authors.forename: Hervé
authors.initials: H
authors.lastname: Ansanay
authors.forename: Maurice
authors.initials: M
authors.lastname: Manning
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.forename: Thierry
authors.initials: T
authors.lastname: Durroux
mesh_terms: D000818:Animals; D065092:Antidiuretic Hormone Receptor Antagonists; D001667:Binding, Competitive; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D005454:Fluorescence Polarization; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D008024:Ligands; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D009942:Organometallic Compounds / Q000737:chemistry; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D011806:Quinolinium Compounds / Q000737:chemistry; D011869:Radioligand Assay; D018045:Receptors, Oxytocin / Q000819:agonists / Q000037:antagonists & inhibitors*; D017483:Receptors, Vasopressin / Q000819:agonists
chemical_list: C481052:Alexa fluor 546; D065092:Antidiuretic Hormone Receptor Antagonists; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008024:Ligands; D009842:Oligopeptides; D009942:Organometallic Compounds; D010456:Peptides, Cyclic; D011806:Quinolinium Compounds; D018045:Receptors, Oxytocin; D017483:Receptors, Vasopressin; C070734:europium(III) trisbipyridine cryptate; C067297:8-aminohexylamino cAMP; D000242:Cyclic AMP