The effects of duloxetine were examined on cloned Kv4.3 channels stably expressed in CHO cells using the whole-cell patch-clamp technique. Duloxetine decreased the peak amplitude of Kv4.3 currents with an acceleration of the decay rate of current inactivation in a concentration-dependent manner. The IC(50) values required for the blocking effects of duloxetine on the peak amplitude and the integral of currents were 8.4 and 2.1μM, respectively. Duloxetine accelerated the rate of inactivation of Kv4.3 currents and thereby decreased the time-to-peak in a concentration-dependent manner. Analysis of the time dependence of the drug block produced estimates of 21.9μM(-1)s(-1) and 165.9s(-1), for the respective association (k(+1)) and dissociation (k(-1)) rate constants. The K(d) value (k(-1)/k(+1)) yielded 7.5μM, which approximates the experimental IC(50) value obtained from the concentration-response curve. The block of Kv4.3 by duloxetine was voltage-dependent at a membrane potential coinciding with the activation of the channels. At a more positive potential, however, the block was relieved. Duloxetine produced a hyperpolarizing shift in the voltage dependence of the steady-state inactivation of Kv4.3, and accelerated the closed-state inactivation of Kv4.3 in the subthreshold voltage range. Duloxetine induced a significant use-dependent block at frequencies of 1 and 2Hz. In the presence of duloxetine, the recovery from inactivation was slower than under control conditions. These results demonstrate that duloxetine exerts a concentration-dependent block of Kv4.3 by binding to the channels in the open and inactivated states and these actions may contribute to its analgesic effect in neuropathic pain.