The activity of zolpidem and other hypnotics within the gamma-aminobutyric acid (GABAA) receptor supramolecular complex, as determined by 35S-t-butylbicyclophosphorothionate (35S-TBPS) binding to rat cerebral cortex membranes.

The present study compares the effects of different hypnotics acting at omega 1/omega 2 sites (zolpidem, zopiclone, flunitrazepam and triazolam) on 35S-t-butylbicyclophosphorothionate (35S-TBPS) binding to well-washed rat cerebral membranes, in the presence of 1M NaCl. Under these conditions, all compounds enhanced 35S-TBPS binding in the 0.05 to 10 microM range with EC50 values and maximal enhancement of: zolpidem, 84 nM and 36%; flunitrazepam, 8 nM and 41%; zopiclone, 171 nM and 51%; triazolam, 2 nM and 42%. Under these conditions, gamma-aminobutyric acid enhanced 35S-TBPS binding with an EC50 of 240 nM and a 38% maximal increase. The EC50 values for the stimulation of 35S-TBPS binding are well correlated, with (r = 0.97) the affinity of these compounds at omega 1/omega 2 sites, and are in the same concentration range. This enhanced binding was due to an altered apparent affinity for the 35S-TBPS recognition site without any change in the number of sites (Scatchard analysis). The effect of zolpidem and other hypnotics was antagonized by flumazenil. This was an apparently competitive antagonism in the case of zolpidem or flunitrazepam, whereas for zopiclone, increasing the concentration of the hypnotic did not overcome the antagonism. Bicuculline only partially antagonized the hypnotic-induced enhancement of 35S-TBPS binding. This antagonism was more effective for zopiclone (-57%) than for either zolpidem (-33%) or flunitrazepam (-30%). Zolpidem and the other hypnotics studied induced a fast component of dissociation which was not observed in the control membranes. These findings are consistent with the hypothesis that omega 1/omega 2 agonists increase the frequency of openings of the chloride ionophore, with both gamma-aminobutyric acid-A receptor-dependent and -independent mechanisms.