Characterization of a new synthetic isoflavonoid with inverse agonist activity at the central benzodiazepine receptor

Abstract

Research aimed at developing selective drugs acting on γ-aminobutyric acid (GABA) A receptors introduced compounds from diverse chemical classes unrelated to the 1,4-benzodiazepines, including flavonoids. These studies also revealed the potential use of inverse agonists as cognition-enhancing agents. Here we report pharmacological properties of the novel synthetic isoflavonoid 2-methoxy-3,8,9-trihydroxy coumestan (PCALC36). PCALC36 displaced [ 3H]flunitrazepam binding to rat brain synaptosomes with an IC 50 of 13.8 μM. Scatchard analysis of the effect of PCALC36 showed a concentration-dependent reduction of the B max of [ 3H]flunitrazepam, without a marked change in K d. This effect could be reversed by diluting and washing the preparation. Addition of 20-μM GABA shifted to the right the inhibition curve of PCALC36 on [ 3H]flunitrazepam binding (IC 50 ratio of 0.68), which is characteristic for inverse agonists. PCALC36 produced little change in the GABAergic tonic currents recorded by whole-cell patch clamp in cultured rat hippocampal neurones, but it caused a 20% reduction in miniature inhibitory postsynaptic current amplitude and completely antagonised the full (direct) agonist midazolam in a quickly reversible manner. The data suggest that the coumestan backbone can be useful for developing novel ligands at the GABA A receptor.