Discovery of a new class of orthosteric antagonists with nanomolar potency at extrasynaptic GABAA receptors
| Autor: | Kasper Harpsøe, Christina Birkedahl Falk-Petersen, Tsonko M. Tsonkov, Uffe Kristiansen, David E. Gloriam, Malene Sofie Nielsen, Petrine Wellendorph, Bente Frølund, Christoffer Bundgaard |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Multidisciplinary 030102 biochemistry & molecular biology Membrane permeability GABAA receptor Chemistry HEK 293 cells lcsh:R Antagonist lcsh:Medicine Pharmacology 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine nervous system Muscimol lcsh:Q Receptor lcsh:Science IC50 030217 neurology & neurosurgery Ionotropic effect |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-12 (2020) Falk-Petersen, C B, Tsonkov, T M, Nielsen, M S, Harpsøe, K, Bundgaard, C, Frølund, B, Kristiansen, U, Gloriam, D E & Wellendorph, P 2020, ' Discovery of a new class of orthosteric antagonists with nanomolar potency at extrasynaptic GABA A receptors ', Scientific Reports, vol. 10, no. 1, 10078 . https://doi.org/10.1038/s41598-020-66821-0 |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-020-66821-0 |
| Popis: | Brain GABAΑ receptors are ionotropic receptors belonging to the class of Cys-loop receptors and are important drug targets for the treatment of anxiety and sleep disorders. By screening a compound library (2,112 compounds) at recombinant human α4β1δ GABAΑ receptors heterologously expressed in a HEK cell line, we identified a scaffold of spirocyclic compounds with nanomolar antagonist activity at GABAΑ receptors. The initial screening hit 2027 (IC50 of 1.03 μM) was used for analogue search resulting in 018 (IC50 of 0.088 μM). 018 was most potent at α3,4,5-subunit containing receptors, thus showing preference for forebrain-expressed extrasynaptic receptors. Schild analysis of 018 at recombinant human α4β1δ receptors and displacement of [3H]muscimol binding in rat cortical homogenate independently confirmed a competitive profile. The antagonist profile of 018 was further validated by whole-cell patch-clamp electrophysiology, where kinetic studies revealed a slow dissociation rate and a shallow hill slope was observed. Membrane permeability studies showed that 2027 and 018 do not cross membranes, thus making the compounds less attractive for studying central GABAΑ receptors effects, but conversely more attractive as tool compounds in relation to emerging peripheral GABAΑ receptor-mediated effects of GABA e.g. in the immune system. |
| Databáze: | OpenAIRE |
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