Open-channel blockers at the human α4β2 neuronal nicotinic acetylcholine receptor

To extend our knowledge of the pharmacological profile of human alpha 4 beta 2 neuronal nicotinic receptors, we investigated the action of hexamethonium on the major brain human nicotinic acetylcholine receptor (nAChR) stably expressed in human embryonic kidney 293 cells. This compound displays all of the characteristics of an open-channel blocker at the human alpha 4 beta 2 nAChR: a voltage-dependent inhibition (more pronounced at hyperpolarized potentials), absence of competition, and use dependence. Moreover, we observed that classic N-methyl-D-aspartate open-channel blockers amantadine, 3,5-dimethyl-1-adamantanamine (memantine), and dizocilpine [(+)-MK-801] and the calcium channel antagonist 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate are powerful inhibitors of the human alpha 4 beta 2 nAChR. Dose-inhibition curves yield, at -100 mV, IC50 values in the micromolar range for all of compounds acid Hill coefficients below unity. Whole-cell current-voltage relationships display a strong rectification profile at hyperpolarized potentials, and current blockades are fitted adequately by a mathematical model that describes the mechanism of an ion channel block. We conclude that these molecules are powerful human alpha 4 beta 2 open-channel blockers ranking in the following order of potency: amantadine = memantine = hexamethonium > 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate similar to MK-801.