Abstract Tracazolate is a pyrazolopyridine anxiolytic that enhances the binding of [ 3H]-flunitrazepam ([ 3H]FLU) to brain tissue. The discovery that a metabolite of tracazolate, desbutyltracazolate, was a weak inhibitor of [ 3H]FLU binding led to the synthesis of a series of potent anxiolytics. From this series, ICI 190,622 emerged as a viable drug candidate, being a potent anxiolytic in rats and monkeys. This anxiolytic agent appears to produce only minimal sedation. Furthermore, ICI 190,622 appears less likely to potentiate the actions of ethanol than diazepam. ICI 190,622 is also a potent anticonvulsant (anti-metrazol ED 50=1.1 mg/kg, PO) in rodents. Neurochemically, ICI 190,622 is similar to the benzodiazepine anxiolytics. In vitro, ICI 190,622 competitively inhibited [ 3H]FLU binding in cerebral cortex with an IC 50 of 81 nM and was 4.3-fold more potent in the cerebellum (IC 50=19 nM). This suggests a selectivity for the Type 1 benzodiazepine binding site. In contrast, diazepam showed similar affinities in both regions (cerebral cortex=7 nM and cerebellum=9 nM). Following oral administration, ICI 190,622 displaced [ 3H]FLU binding from cerebellar membranes more potently than diazepam (ED 50=3 and 6 mg/kg, respectively, 1 hour after administration). Thus, ICI 190,622 should be an effective anxiolytic with significant advantages over benzodiazepines.