Fluorescent ligands for dopamine D2/D3 receptors

Sci Rep. 2020 Dec 14;10(1):21842. doi: 10.1038/s41598-020-78827-9.

Abstract

Fluorescent ligands are versatile tools for the study of G protein-coupled receptors. Depending on the fluorophore, they can be used for a range of different applications, including fluorescence microscopy and bioluminescence or fluorescence resonance energy transfer (BRET or FRET) assays. Starting from phenylpiperazines and indanylamines, privileged scaffolds for dopamine D2-like receptors, we developed dansyl-labeled fluorescent ligands that are well accommodated in the binding pockets of D2 and D3 receptors. These receptors are the target proteins for the therapy for several neurologic and psychiatric disorders, including Parkinson's disease and schizophrenia. The dansyl-labeled ligands exhibit binding affinities up to 0.44 nM and 0.29 nM at D2R and D3R, respectively. When the dansyl label was exchanged for sterically more demanding xanthene or cyanine dyes, fluorescent ligands 10a-c retained excellent binding properties and, as expected from their indanylamine pharmacophore, acted as agonists at D2R. While the Cy3B-labeled ligand 10b was used to visualize D2R and D3R on the surface of living cells by total internal reflection microscopy, ligand 10a comprising a rhodamine label showed excellent properties in a NanoBRET binding assay at D3R.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Carbocyanines / chemistry*
  • Cricetulus
  • Fluorescent Dyes / chemistry*
  • HEK293 Cells
  • Humans
  • Receptors, Dopamine D2* / chemistry
  • Receptors, Dopamine D2* / genetics
  • Receptors, Dopamine D2* / metabolism
  • Receptors, Dopamine D3* / chemistry
  • Receptors, Dopamine D3* / genetics
  • Receptors, Dopamine D3* / metabolism

Substances

  • Carbocyanines
  • DRD2 protein, human
  • DRD3 protein, human
  • Fluorescent Dyes
  • Receptors, Dopamine D2
  • Receptors, Dopamine D3
  • cyanine dye 3