Enhancement of benzodiazepine and GABA binding by the novel anxiolytic, tracazolate

Tracazolate (ICI 136,753) 4-butylamine-1-ethyl-6-methyl-1H-pyrazolo[3,4]pyridine-5-carboxylic acid ethyl ester is a non-benzodiazepine with anxiolytic-like activity in animal models. In contrast to the benzodiazepines, it enhances [ 3H]flunitrazepam binding in rat synaptic membrane fragments. The enhancement is potentiated by chloride ion and is due to an increase in affinity of the receptor. The enhancement of benzodiazepine binding by γ-aminobutyric acid (GABA) is additive with that of tracazolate; however, the GABA antagonist bicuculline blocks the enhancement by both compounds. Tracazolate enhances [ 3H]GABA binding to frozen and thawed Triton X-100-treated membrane fragments. The enhancement is due to an increase in the number of sites and potentiated by chloride. Benzodiazepines also enhanced GABA binding but the effect was due to an apparent change in affinity and not potentiated by chloride. The rank order of chlordiazepoxide, diazepam and flunitrazepam for enhancement of GABA binding and displacement of [ 3H]flunitrazepam binding were the same. The enhancement of [ 3H]GABA binding by flunitrazepam and tracazolate were additive. Possible interactions between these various receptors are discussed.