Electrophysiological characterization of 14-benzoyltalatisamine, a selective blocker of the delayed rectifier K + channel found in virtual screening

Abstract

14-Benzoyltalatisamine is a potent and selective blocker of the delayed rectifier K + channel found in a computational virtual screening study. The compound was found to block the K + channel from the extracellular side. However, it is unclear whether 14-benzoyltalatisamine shares the same block mechanism with tetraethylammonium (TEA). In order to elucidate how the hit compound found by the virtual screening interacts with the outer vestibule of the K + channel, the effects of 14-benzoyltalatisamine and TEA on the delayed rectifier K + current of rat dissociated hippocampal neurons were compared using whole-cell voltage-clamp recording. External application of 14-benzoyltalatisamine and TEA reversibly inhibited the current with IC 50 values of 10.1 ± 2.2 μM and 1.05 ± 0.21 mM, respectively. 14-Benzoyltalatisamine exerted voltage-dependent inhibition, markedly accelerated the decay of the current, and caused a significant hyperpolarizing shift of the steady-state activation curve, whereas TEA caused voltage-independent inhibition, without affecting the kinetic parameters of the current. The blockade by 14-benzoyltalatisamine, but not by TEA, was significantly diminished in a high K + (60 mM) external solution. The potency of 14-benzoyltalatisamine was markedly reduced in the presence of 15 mM TEA. The results suggest that 14-benzoyltalatisamine bind to the external pore entry of the delayed rectifier K + channel with partial insertion into the selectivity filter, which is in conformity with that predicted by the molecular docking model in the virtual screening.