© 2004 by European Society of Cardiology
Copyright © 2004, European Society of Cardiology
Diltiazem inhibits hKv1.5 and Kv4.3 currents at therapeutic concentrations
Department of Pharmacology, School of Medicine, Universidad Complutense, 28040 Madrid, Spain
* Corresponding author. Tel.: +34 91 394 14 74; fax: +34 91 394 14 70. Email address: rcaballero{at}ift.csic.es
Objective: In the present study we examined the effects of diltiazem, an L-type Ca2+ channel blocker widely used for the control of the ventricular rate in patients with supraventricular arrhythmias, on hKv1.5 and Kv4.3 channels that generate the cardiac ultrarapid delayed rectifier (IKur) and the 4-aminopyridine sensitive transient outward (Ito) K+ currents, respectively.
Methods: hKv1.5 and Kv4.3 channels were stably and transiently expressed in mouse fibroblast and Chinese hamster ovary cells, respectively. Currents were recorded using the whole-cell patch clamp.
Results: Diltiazem (0.01 nM-500 µM) blocked hKv1.5 channels, in a frequency-dependent manner exhibiting a biphasic dose-response curve (IC50=4.8±1.5 nM and 42.3±3.6 µM). Diltiazem delayed the initial phase of the tail current decline and shifted the midpoint of the activation (Vh=–16.5±2.1 mV vs –20.4±2.6 mV, P<0.001) and inactivation (Vh=–22.4±0.7 mV vs. –28.2±1.9 mV, P<0.001) curves to more negative potentials. The analysis of the development of the diltiazem-induced block yielded apparent association (k) and dissociation (P) rate constants of (1.6±0.2)x106 M–1s–1 and 46.8±4.8 s–1, respectively. Diltiazem (0.1 nM-100 µM) also blocked Kv4.3 channels in a frequency-dependent manner exhibiting a biphasic dose-response curve (IC50=62.6±11.1 nM and 109.9±12.8 µM). Diltiazem decreased the peak current and, at concentrations 
0.1 µM, accelerated the inactivation time course. The apparent association and dissociation rate constants resulted (1.7±0.2)x106 M–1s–1 and 258.6±38.1 s–1, respectively. Diltiazem, 10 nM, shifted to more negative potentials the voltage-dependence of Kv4.3 channel inactivation (Vh=–33.1±2.3 mV vs –38.2±3.5 mV, n=6, Plt;0.05) the blockade increasing at potentials at which the amount of inactivated channels increased.
Conclusion: The results demonstrated for the first time that diltiazem, at therapeutic concentrations, decreased hKv1.5 and Kv4.3 currents by binding to the open and the inactivated state of the channels.
1 These authors contributed equally to this work.