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

In the mammalian central nervous system (CNS) GABA_A receptors (GABA_ARs) mediate neuronal inhibition and are important therapeutic targets. GABA_ARs are composed of 5 subunits, drawn from 19 proteins, underpinning expression of 20-30 GABA_AR 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 GABA_ARs (α5-GABA_ARs). Current α5-GABA_AR inhibitors bind to the "benzodiazepine site". However, in HEK293 cells expressing recombinant α5-GABA_ARs, S44819 had no effect on ³H-flumazenil binding, but displaced the GABA_AR 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-GABA_ARs, but had no effect on synaptic GABA_ARs. In mouse thalamic neurons, S44819 had no effect on the tonic current mediated by δ-GABA_ARs, or on synaptic (α1β2γ2) GABA_ARs. 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-GABA_ARs. 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.