title: Analysis of β-Subunit-dependent GABAA Receptor Modulation and Behavioral Effects of Valerenic Acid Derivatives.
abstract: Valerenic acid (VA)-a β2/3-selective GABA type A (GABAA) receptor modulator-displays anxiolytic and anticonvulsive effects in mice devoid of sedation, making VA an interesting drug candidate. Here we analyzed β-subunit-dependent enhancement of GABA-induced chloride currents (IGABA) by a library of VA derivatives and studied their effects on pentylenetetrazole (PTZ)-induced seizure threshold and locomotion. Compound-induced IGABA enhancement was determined in oocytes expressing α1β1γ2S, α1β2γ2S, or α1β3γ2S receptors. Effects on seizure threshold and locomotion were studied using C57BL/6N mice and compared with saline-treated controls. β2/3-selective VA derivatives such as VA-amide (VA-A) modulating α1β3γ2S (VA-A: Emax = 972 ± 69%, n = 6, P < 0.05) and α1β2γ2S receptors (Emax = 1119 ± 72%, n = 6, P < 0.05) more efficaciously than VA (α1β3γ2S: VA: Emax = 632 ± 88%, n = 9 versus α1β2γ2S: VA: Emax = 721 ± 68%, n = 6) displayed significantly more pronounced seizure threshold elevation than VA (saline control: 40.4 ± 1.4 mg/kg PTZ versus VA 10 mg/kg: 49.0 ± 1.8 mg/kg PTZ versus VA-A 3 mg/kg: 57.9 ± 1.9 mg/kg PTZ, P < 0.05). Similarly, VA's methylamide (VA-MA) enhancing IGABA through β3-containing receptors more efficaciously than VA (Emax = 1043 ± 57%, P < 0.01, n = 6) displayed stronger anticonvulsive effects. Increased potency of IGABA enhancement and anticonvulsive effects at lower doses compared with VA were observed for VA-tetrazole (α1β3γ2S: VA-TET: EC50 = 6.0 ± 1.0 μM, P < 0.05; VA-TET: 0.3 mg/kg: 47.3 ± 0.5 mg/kg PTZ versus VA: 10 mg/kg: 49.0 ± 1.8 mg/kg PTZ, P < 0.05). At higher doses (≥10 mg/kg), VA-A, VA-MA, and VA-TET reduced locomotion. In contrast, unselective VA derivatives induced anticonvulsive effects only at high doses (30 mg/kg) or did not display any behavioral effects. Our data indicate that the β2/3-selective compounds VA-A, VA-MA, and VA-TET induce anticonvulsive effects at low doses (≤10 mg/kg), whereas impairment of locomotion was observed at doses ≥10 mg/kg.
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.) sophia.khom@univie.ac.at schwarzer.christoph@i-med.ac.at.
authors.forename: S
authors.initials: S
authors.lastname: Khom
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: J
authors.initials: J
authors.lastname: Hintersteiner
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: D
authors.initials: D
authors.lastname: Luger
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: M
authors.initials: M
authors.lastname: Haider
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: G
authors.initials: G
authors.lastname: Pototschnig
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: M D
authors.initials: MD
authors.lastname: Mihovilovic
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: C
authors.initials: C
authors.lastname: Schwarzer
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Vienna, Austria (S.K., J.H., D.L., S.H.); Institute of Applied Synthetic Chemistry, TU Wien, Vienna, Austria (M.H., G.P., M.D.M.); and Department of Pharmacology, Medical University of Innsbruck, Innsbruck, Austria (C.S.).
authors.forename: S
authors.initials: S
authors.lastname: Hering
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005260:Female; D007192:Indenes / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D051379:Mice; D010433:Pentylenetetrazole / Q000009:adverse effects; D011963:Receptors, GABA-A / Q000378:metabolism*; D012640:Seizures / Q000139:chemically induced / Q000188:drug therapy; D012717:Sesquiterpenes / Q000737:chemistry* / Q000494:pharmacology* / Q000627:therapeutic use; D014982:Xenopus laevis
chemical_list: D007192:Indenes; D011963:Receptors, GABA-A; D012717:Sesquiterpenes; C037133:valerenic acid; D010433:Pentylenetetrazole