HIV Protease Inhibitors: Suppression of Insulin Secretion by Inhibition of Voltage-Dependent K+Currents and Anion Currents

Abstract

We have shown before that the human immunodeficiency virus (HIV) protease inhibitors ritonavir and nelfinavir, but not indinavir, suppress insulin secretion from mouse pancreatic B-cells via reduction of the cytosolic free calcium concentration ([Ca(2+)](c)). This was not because of an effect on ATP-dependent K(+) channels (K(ATP) channels) or L-type Ca(2+) channels. The study was intended to elucidate the mechanisms by which distinct HIV protease inhibitors decrease [Ca(2+)](c) and thus evoke their adverse side effect on insulin release. Membrane potential and whole-cell currents were measured with the patch-clamp technique, and [Ca(2+)](c) was determined with a fluorescence dye. Ritonavir and nelfinavir both inhibited the same component(s) of voltage-dependent K(+) currents with a concomitant change in action potential wave form, whereas indinavir was ineffective. Comparison with other blockers of voltage-dependent K(+) currents revealed that suppression of distinct noninactivating current component(s) altered action potential wave form and decreased [Ca(2+)](c) similar to ritonavir and nelfinavir, whereas blockage of inactivating component(s) was without effect. Complete inhibition of voltage-dependent K(+) currents by 80 mM TEA(+) drastically increased [Ca(2+)](c), demonstrating that voltage-dependent K(+) channels are not the sole target of ritonavir and nelfinavir. Accordingly, the Ca(2+)-lowering effect of ritonavir was preserved in the presence of 80 mM TEA(+). This effect was mimicked by the anion channel blocker 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Consequentially, ritonavir and nelfinavir inhibited a DIDS-sensitive anion current in B-cells. We suggest that ritonavir and nelfinavir decrease insulin secretion by inhibition of voltage-dependent K(+) channels and anion channels, which are essential to provide counterion currents for Ca(2+) influx across the plasma membrane.