Selective fluorescent nonpeptidic antagonists for vasopressin V₂ GPCR: application to ligand screening and oligomerization assays

J Med Chem. 2012 Oct 25;55(20):8588-602. doi: 10.1021/jm3006146. Epub 2012 Oct 5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antidiuretic Hormone Receptor Antagonists*
  • Benzazepines / chemical synthesis*
  • Benzazepines / chemistry
  • Benzazepines / pharmacology
  • Click Chemistry
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes / chemical synthesis*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / pharmacology
  • High-Throughput Screening Assays
  • Ligands
  • Models, Molecular
  • Protein Multimerization
  • Receptors, Oxytocin / antagonists & inhibitors
  • Receptors, Oxytocin / metabolism
  • Receptors, Vasopressin / metabolism*
  • Solubility
  • Structure-Activity Relationship

Substances

  • Antidiuretic Hormone Receptor Antagonists
  • Benzazepines
  • Fluorescent Dyes
  • Ligands
  • Receptors, Oxytocin
  • Receptors, Vasopressin