Selective inhibition of extra-synaptic α5-GABAA receptors by S44819, a new therapeutic agent

Neuropharmacology. 2017 Oct:125:353-364. doi: 10.1016/j.neuropharm.2017.08.012. Epub 2017 Aug 12.

Abstract

In the mammalian central nervous system (CNS) GABAA receptors (GABAARs) mediate neuronal inhibition and are important therapeutic targets. GABAARs are composed of 5 subunits, drawn from 19 proteins, underpinning expression of 20-30 GABAAR subtypes. In the CNS these isoforms are heterogeneously expressed and exhibit distinct physiological and pharmacological properties. We report the discovery of S44819, a novel tricyclic oxazolo-2,3-benzodiazepine-derivative, that selectively inhibits α5-subunit-containing GABAARs (α5-GABAARs). Current α5-GABAAR inhibitors bind to the "benzodiazepine site". However, in HEK293 cells expressing recombinant α5-GABAARs, S44819 had no effect on 3H-flumazenil binding, but displaced the GABAAR agonist 3H-muscimol and competitively inhibited the GABA-induced responses. Importantly, we reveal that the α5-subunit selectivity is uniquely governed by amino acid residues within the α-subunit F-loop, a region associated with GABA binding. In mouse hippocampal CA1 neurons, S44819 enhanced long-term potentiation (LTP), blocked a tonic current mediated by extrasynaptic α5-GABAARs, but had no effect on synaptic GABAARs. In mouse thalamic neurons, S44819 had no effect on the tonic current mediated by δ-GABAARs, or on synaptic (α1β2γ2) GABAARs. In rats, S44819 enhanced object recognition memory and reversed scopolamine-induced impairment of working memory in the eight-arm radial maze. In conclusion, S44819 is a first in class compound that uniquely acts as a potent, competitive, selective antagonist of recombinant and native α5-GABAARs. Consequently, S44819 enhances hippocampal synaptic plasticity and exhibits pro-cognitive efficacy. Given this profile, S44819 may improve cognitive function in neurodegenerative disorders and facilitate post-stroke recovery.

Keywords: Cognition; Extrasynaptic GABA(A) receptors; Long term potentiation; Phasic inhibition; Tonic inhibition; α5-GABA(A) receptors.

MeSH terms

  • Animals
  • Benzodiazepines / pharmacology*
  • Binding, Competitive
  • Brain / drug effects
  • Brain / metabolism
  • Female
  • Flumazenil / pharmacology
  • GABA-A Receptor Agonists / pharmacology
  • GABA-A Receptor Antagonists / pharmacology*
  • HEK293 Cells
  • Humans
  • Long-Term Potentiation / drug effects
  • Long-Term Potentiation / physiology
  • Male
  • Memory / drug effects
  • Memory / physiology
  • Mice, Inbred C57BL
  • Muscimol / pharmacology
  • Neural Inhibition / drug effects
  • Neural Inhibition / physiology
  • Nootropic Agents / pharmacology*
  • Oxazoles / pharmacology*
  • Rats, Sprague-Dawley
  • Receptors, GABA-A / metabolism*
  • Tissue Culture Techniques
  • gamma-Aminobutyric Acid / pharmacology

Substances

  • GABA-A Receptor Agonists
  • GABA-A Receptor Antagonists
  • GABRA5 protein, human
  • Gabra5 protein, mouse
  • Gabra5 protein, rat
  • Nootropic Agents
  • Oxazoles
  • Receptors, GABA-A
  • S44819
  • Benzodiazepines
  • Muscimol
  • Flumazenil
  • gamma-Aminobutyric Acid