Binding Characteristics and Interactions of Depressant Drugs with [³⁵S]t‐Butylbicyclophosphorothionate, a Ligand that Binds to the Picrotoxinin Site

Abstract: [³⁵S]r‐Butylbicyclophosphorothionate (TBPT), a cage convulsant with picrotoxinin‐like activity, binds to rat brain membranes to a single site with an apparent K_D of 25.1 ± 5.6 nM and a B_max of 1.40 ± 0.22 pmol/mg protein. TBPT binding to rat brain membranes was inhibited by a variety of convulsant, depressant, anxiolytic, and anticonvulsant drugs that had previously been shown to inhibit [³H]a‐dihydropicrotoxinin binding. Depressant drugs such as pentobarbital and the nonbarbiturate (+)etomidate inhibited TBPT binding in an uncompetitive manner. Thus, pentobarbital and (+)etomidate decreased both the affinity and the number of binding sites of TBPT to whole brain membranes. The IC₅₀ values of (+)etomidate (9 μM) and pentobarbital (90 μM) are similar to the EC₅₀ values at which they enhance both [³H]‐γ‐aminobutyric acid and [³H]diazepam binding in cerebral cortex membranes. RO5–4864, which has recently been shown to be a convulsant, also inhibited TBPT binding (IC₅₀= 10 μM). These results suggest that TBPT binds to the picrotoxinin site and further supports the notion that the picrotoxinin site is an important modulatory site at the benzodiazepine‐GABA receptor‐ionophore complex.