Structure-based discovery of antagonists for GluN3-containing N-methyl-D-aspartate receptors

• Type of Material: Article
• Authors: Trine Kvist, University of Copenhagen; Jeremy R. Greenwood, Schrodinger Inc.; Kasper B. Hansen, Emory University; Stephen Traynelis, Emory University; Hans Brauner-Osborne, University of Copenhagen
• Language: English
• Date: 2013-12-01
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2013 Elsevier Ltd. All rights reserved. CC BY NC ND 4.0.
• Final Published Version (URL): http://dx.doi.org/10.1016/j.neuropharm.2013.08.003
• Title of Journal or Parent Work: Neuropharmacology
• ISSN: 0028-3908
• Volume: 75
• Start Page: 324
• End Page: 336
• Grant/Funding Information: This work was supported by the GluTarget Programme of Excellence at the University of Copenhagen, the Danish Ministry of Science, Innovation and Higher Education’s EliteForsk Programme and NIH-NINS (N036654, NS065371 SFT).
• Keywords: NMDA RECEPTOR; PERMEABILITY; GluN3 subunit; INHIBITION; Virtual screening; Xenopus oocyte electrophysiology; GLYCINE-BINDING; Selectivity; Neurosciences; NR3A-CONTAINING NMDARS; NR1; EXPRESSION; Antagonist; NR3A SUBUNIT; NMDA receptor; Life Sciences & Biomedicine; Neurosciences & Neurology; Science & Technology; Pharmacology & Pharmacy; ACTIVATION; LOCALIZATION
• Research Categories: Health Sciences, Pharmacology

Abstract

NMDA receptors are ligand-gated ion channels that assemble into tetrameric receptor complexes composed of glycine-binding GluN1 and GluN3 subunits (GluN3A-B) and glutamate-binding GluN2 subunits (GluN2A-D). NMDA receptors can assemble as GluN1/N2 receptors and as GluN3-containing NMDA receptors, which are either glutamate/glycine-activated triheteromeric GluN1/N2/N3 receptors or glycine-activated diheteromeric GluN1/N3 receptors. The glycine-binding GluN1 and GluN3 subunits display strikingly different pharmacological selectivity profiles. However, the pharmacological characterization of GluN3-containing receptors has been hampered by the lack of methods and pharmacological tools to study GluN3 subunit pharmacology in isolation. Here, we have developed a method to study the pharmacology of GluN3 subunits in recombinant diheteromeric GluN1/N3 receptors by mutating the orthosteric ligand-binding pocket in GluN1. This method is suitable for performing compound screening and characterization of structure-activity relationship studies on GluN3 ligands. We have performed a virtual screen of the orthosteric binding site of GluN3A in the search for antagonists with selectivity for GluN3 subunits. In the subsequent pharmacological evaluation of 99 selected compounds, we identified 6-hydroxy-[1,2,5]oxadiazolo[3,4-b]pyrazin-5(4H)-one (TK80) a novel competitive antagonist with preference for the GluN3B subunit. Serendipitously, we also identified [2-hydroxy-5-((4-(pyridin-3-yl)thiazol-2-yl)amino]benzoic acid (TK13) and 4-(2,4-dichlorobenzoyl)-1H-pyrrole-2-carboxylic acid (TK30), two novel non-competitive GluN3 antagonists. These findings demonstrate that structural differences between the orthosteric binding site of GluN3 and GluN1 can be exploited to generate selective ligands.