Abstract

A uricosuric drug benzbromarone, widely used for treatment of gout, hyperpolarizes the membrane potential of airway smooth muscle cells, but how it works remains unknown. Here we show a novel role of benzbromarone in activation of large conductance calcium-activated K⁺ channels. Benzbromarone results in dose-dependent activation of macroscopic BK currents about 1.7 to 14.5 folds with an EC₅₀ of 111 μM and shifts the voltage-dependent channel activation to a more hyperpolarizing direction about 10 to 54 mV in whole-cell patch clamp recordings. In single-channel recordings, benzbromarone decreases single BKα channel closed dwell time and increases the channel open probability. Co-expressing β1 subunit also enhances BK activation by benzbromarone with an EC50 of 67 μM and a leftward shift of G-V curve about 44 to 138 mV. Site-directed mutagenesis reveals that a motif of three amino acids ³²⁹RKK³³¹ in the cytoplasmic linker between S6 and C-terminal RCK gating ring mediates the pharmacological activation of BK channels by benzbromarone. Further ex vivo assay shows that benzbromarone causes reduction of tracheal strip contraction. Taken together, our findings demonstrate that uricosuric benzbromarone activates BK channels through molecular mechanism of action involving the channel RKK motif of S6-RCK linker. Pharmacological activation of BK channel by benzbromarone causes reduction of tracheal strip contraction, holding a repurposing potential for asthma and pulmonary arterial hypertension or BK channelopathies.

Significance Statement We describe a novel role of uricosuric agent benzbromarone in BK channel activation and relaxation of airway smooth muscle contraction. In this study, we find that benzbromarone is an activator of the large-conductance Ca²⁺- and voltage-activated K⁺ channel (BK channel) that serves numerous cellular functions including control of smooth muscle contraction. Pharmacological activation of BK channel by an FDA approved drug benzbromarone may hold repurposing potential for treatment of asthma and pulmonary arterial hypertension or BK channeopathies.