Discovery of a new class of orthosteric antagonists with nanomolar potency at extrasynaptic GABA A receptors
Brain GABA 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 α β δ GABA receptors heterologously expressed in a HEK ce...
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Published in | Scientific reports Vol. 10; no. 1; p. 10078 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
22.06.2020
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Abstract | Brain GABA
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 α
β
δ GABA
receptors heterologously expressed in a HEK cell line, we identified a scaffold of spirocyclic compounds with nanomolar antagonist activity at GABA
receptors. The initial screening hit 2027 (IC
of 1.03 μM) was used for analogue search resulting in 018 (IC
of 0.088 μM). 018 was most potent at α
-subunit containing receptors, thus showing preference for forebrain-expressed extrasynaptic receptors. Schild analysis of 018 at recombinant human α
β
δ receptors and displacement of [
H]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 GABA
receptors effects, but conversely more attractive as tool compounds in relation to emerging peripheral GABA
receptor-mediated effects of GABA e.g. in the immune system. |
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AbstractList | Brain GABA
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 α
β
δ GABA
receptors heterologously expressed in a HEK cell line, we identified a scaffold of spirocyclic compounds with nanomolar antagonist activity at GABA
receptors. The initial screening hit 2027 (IC
of 1.03 μM) was used for analogue search resulting in 018 (IC
of 0.088 μM). 018 was most potent at α
-subunit containing receptors, thus showing preference for forebrain-expressed extrasynaptic receptors. Schild analysis of 018 at recombinant human α
β
δ receptors and displacement of [
H]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 GABA
receptors effects, but conversely more attractive as tool compounds in relation to emerging peripheral GABA
receptor-mediated effects of GABA e.g. in the immune system. |
Author | Kristiansen, Uffe Gloriam, David E Wellendorph, Petrine Nielsen, Malene Sofie Frølund, Bente Tsonkov, Tsonko M Harpsøe, Kasper Bundgaard, Christoffer Falk-Petersen, Christina Birkedahl |
Author_xml | – sequence: 1 givenname: Christina Birkedahl surname: Falk-Petersen fullname: Falk-Petersen, Christina Birkedahl organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 2 givenname: Tsonko M surname: Tsonkov fullname: Tsonkov, Tsonko M organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 3 givenname: Malene Sofie surname: Nielsen fullname: Nielsen, Malene Sofie organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 4 givenname: Kasper surname: Harpsøe fullname: Harpsøe, Kasper organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 5 givenname: Christoffer surname: Bundgaard fullname: Bundgaard, Christoffer organization: Translational DMPK, H. Lundbeck A/S, Ottiliavej 9, 2500, Valby, Denmark – sequence: 6 givenname: Bente surname: Frølund fullname: Frølund, Bente organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 7 givenname: Uffe surname: Kristiansen fullname: Kristiansen, Uffe organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 8 givenname: David E surname: Gloriam fullname: Gloriam, David E organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark – sequence: 9 givenname: Petrine surname: Wellendorph fullname: Wellendorph, Petrine email: pw@sund.ku.dk organization: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen Ø, Denmark. pw@sund.ku.dk |
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receptors are ionotropic receptors belonging to the class of Cys-loop receptors and are important drug targets for the treatment of anxiety and... |
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SubjectTerms | Drug Evaluation, Preclinical - methods Electrophysiological Phenomena - drug effects GABA-A Receptor Antagonists - classification GABA-A Receptor Antagonists - isolation & purification GABA-A Receptor Antagonists - pharmacology gamma-Aminobutyric Acid - metabolism HEK293 Cells Humans Kinetics Membrane Potentials - drug effects Patch-Clamp Techniques Protein Subunits - metabolism Receptors, GABA-A - metabolism Small Molecule Libraries - pharmacology |
Title | Discovery of a new class of orthosteric antagonists with nanomolar potency at extrasynaptic GABA A receptors |
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