Differences in kinetics of structurally related competitive GABAA receptor antagonists

Previously, 4-alkyl and 4-aryl substituted analogues of the low-efficacy partial GABAA receptor agonist 5-(4-piperidyl)-3-isothiazole (4-PIOL) have been identified as competitive GABAA receptor antagonists. These structurally related competitive antagonists show marked differences in their kinetic properties. The kinetics of 20 4-alkyl and 4-aryl substituted analogues of 4-PIOL, two 4-arylalkyl substituted 3-isothiazolol analogues and the classical GABAA receptor antagonist SR95531 was studied in cultured cerebral cortical neurons using whole-cell patch-clamp techniques. The kinetics of the antagonists was studied indirectly by measuring the changes in the response of the full GABAA receptor agonist isoguvacine (IGU) induced by concurrent application of an antagonist. When added, the majority of the antagonists did not affect the rate of deactivation of the IGU-induced responses. When removed, however, the majority of the antagonists slowed the reactivation phase of IGU implying that the dissociation of the antagonist from the GABAA receptor is the rate-limiting step. Surprisingly, the functional off-rates of the antagonists seemed to correlate better with the lipophilicity of the compounds than with the affinity and potency. This suggests that the dissociation of the tested antagonists from the GABAA receptor is restricted by lipophilic interactions, perhaps with the aromatic amino acids surrounding the GABA binding site.