2'-Methoxy-6-methylflavone: a novel anxiolytic and sedative with subtype selective activating and modulating actions at GABA(A) receptors.

BACKGROUND AND
PURPOSE: Flavonoids are known to have anxiolytic and sedative effects mediated via actions on ionotropic GABA receptors. We sought to investigate this further. EXPERIMENTAL APPROACH: We evaluated the effects of 2'-methoxy-6-methylflavone (2'MeO6MF) on native GABA(A) receptors in new-born rat hippocampal neurons and determined specificity from 18 human recombinant GABA(A) receptor subtypes expressed in Xenopus oocytes. We used ligand binding, two-electrode voltage clamp and patch clamp studies together with behavioural studies. KEY
RESULTS: 2'MeO6MF potentiated GABA at α2β1γ2L and all α1-containing GABA(A) receptor subtypes. At α2β2/3γ2L GABA(A) receptors, however, 2'MeO6MF directly activated the receptors without potentiating GABA. This activation was attenuated by bicuculline and gabazine but not flumazenil indicating a novel site. Mutation studies showed position 265 in the β1/2 subunit was key to whether 2'MeO6MF was an activator or a potentiator. In hippocampal neurons, 2'MeO6MF directly activated single-channel currents that showed the hallmarks of GABA(A) Cl(-) currents. In the continued presence of 2'MeO6MF the single-channel conductance increased and these high conductance channels were disrupted by the γ2(381-403) MA peptide, indicating that such currents are mediated by α2/γ2-containing GABA(A) receptors. In mice, 2'MeO6MF (1-100 mg·kg(-1) ; i.p.) displayed anxiolytic-like effects in two unconditioned models of anxiety: the elevated plus maze and light/dark tests. 2'MeO6MF induced sedative effects at higher doses in the holeboard, actimeter and barbiturate-induced sleep time tests. No myorelaxant effects were observed in the horizontal wire test. CONCLUSIONS AND IMPLICATIONS: 2'MeO6MF will serve as a tool to study the complex nature of the activation and modulation of GABA(A) receptor subtypes.