The purpose of this study was to evaluate the electrophysiologic effects of zatebradine, a sinus node inhibitor, in human subjects. Patients without structural heart disease were randomized to receive intravenous zatebradine (23 patients) or placebo (12 patients). Electrophysiologic measures were obtained at baseline and repeated at 40 and 70 min after drug administration. In the placebo group, there were no significant changes in any parameter over time. After zatebradine, sinus node function changed significantly at 40 min, with no further change at 70 min; sinus cycle length was prolonged by 16 and 17% (p < 0.001), and corrected sinus node recovery time was prolonged by 30 and 22% (p = 0.008). Similarly, atrioventricular node function changed significantly at 40 min, with no further change at 70 min; atrio-His interval was prolonged by 15 and 15% (p = 0.02), atrioventricular node effective refractory period was prolonged by 12 and 11% (p = 0.01), and Wenckebach cycle length was prolonged by 15 and 11% (p = 0.002). Atrial refractoriness, His-Purkinje conduction, ventricular refractoriness, and action-potential duration were not affected by zatebradine. Zatebradine, a sinus node inhibitor, alters the conduction and refractory properties of the human atrioventricular node, in addition to the expected effect on sinus node function.