Simultaneous Multiple MS Binding Assays for the Dopamine, Norepinephrine, and Serotonin Transporters

ChemMedChem. 2018 Mar 6;13(5):453-463. doi: 10.1002/cmdc.201700737. Epub 2018 Feb 16.

Abstract

In this work, we present label-free, mass-spectrometry-based binding assays (MS Binding Assays), targeting the human dopamine, norepinephrine, and serotonin transporters (hDAT, hNET, and hSERT) in simultaneous binding experiments. Using a validated LC-ESI-MS/MS method for quantification of the selective dopamine transporter inhibitor (R,R)-4-(2-benzhydryloxyethyl)-1-(4-fluorobenzyl)piperidin-3-ol ((R,R)-D-84), the selective norepinephrine transporter inhibitor (S,S)-reboxetine, and the selective serotonin reuptake inhibitor (S)-citalopram, binding affinities at the three monoamine transporters could be characterized simultaneously in a single binding experiment. The performed simultaneous saturation and competition experiments yielded results that are in good accordance with those determined in MS Binding Assays addressing the monoamine transporters individually. The results obtained from this study underscore the potential of MS Binding Assays for simultaneous affinity determination at different targets, which is difficult to accomplish with conventional radioligand binding assays.

Keywords: binding assays; liquid chromatography; mass spectrometry; monoamine transporters; neurotransmitters.

MeSH terms

  • Benzhydryl Compounds / pharmacology*
  • Binding Sites / drug effects
  • Chromatography, Liquid
  • Citalopram / pharmacology*
  • Dopamine Plasma Membrane Transport Proteins / antagonists & inhibitors*
  • Dopamine Plasma Membrane Transport Proteins / metabolism
  • Humans
  • Molecular Structure
  • Norepinephrine Plasma Membrane Transport Proteins / antagonists & inhibitors*
  • Norepinephrine Plasma Membrane Transport Proteins / metabolism
  • Piperidines / pharmacology*
  • Reboxetine / pharmacology*
  • Serotonin Plasma Membrane Transport Proteins / metabolism*
  • Spectrometry, Mass, Electrospray Ionization
  • Tandem Mass Spectrometry

Substances

  • 4-(2-benzhydryloxyethyl)-1-(4-fluorobenzyl)piperidin-3-ol
  • Benzhydryl Compounds
  • Dopamine Plasma Membrane Transport Proteins
  • Norepinephrine Plasma Membrane Transport Proteins
  • Piperidines
  • Serotonin Plasma Membrane Transport Proteins
  • Citalopram
  • Reboxetine