Korean J Physiol Pharmacol.  2022 Sep;26(5):397-404. 10.4196/kjpp.2022.26.5.397.

Inhibition of voltage-dependent K⁺ channels by antimuscarinic drug fesoterodine in coronary arterial smooth muscle cells

• Park S
• Kang M
• Heo R
• Mun SY
• Park M
• Han ET
• Han JH
• Chun W
• Park H
• Park WS

• KMID: 2532769
• DOI: http://doi.org/10.4196/kjpp.2022.26.5.397

Abstract

Fesoterodine, an antimuscarinic drug, is widely used to treat overactive bladder syndrome. However, there is little information about its effects on vascular K⁺ channels. In this study, voltage-dependent K⁺ (Kv) channel inhibition by fesoterodine was investigated using the patch-clamp technique in rabbit coronary artery. In whole-cell patches, the addition of fesoterodine to the bath inhibited the Kv currents in a concentration-dependent manner, with an IC₅₀ value of 3.19 ± 0.91 μM and a Hill coefficient of 0.56 ± 0.03. Although the drug did not alter the voltage-dependence of steady-state activation, it shifted the steady-state inactivation curve to a more negative potential, suggesting that fesoterodine affects the voltage-sensor of the Kv channel. Inhibition by fesoterodine was significantly enhanced by repetitive train pulses (1 or 2 Hz). Furthermore, it significantly increased the recovery time constant from inactivation, suggesting that the Kv channel inhibition by fesoterodine is use (state)-dependent. Its inhibitory effect disappeared by pretreatment with a Kv 1.5 inhibitor. However, pretreatment with Kv2.1 or Kv7 inhibitors did not affect the inhibitory effects on Kv channels. Based on these results, we conclude that fesoterodine inhibits vascular Kv channels (mainly the Kv1.5 subtype) in a concentration- and use (state)-dependent manner, independent of muscarinic receptor antagonism.

Keyword

Coronary artery; Fesoterodine; Kv potassium channel; Kv1.5 potassium channel; Use-dependence

Figure

• 

  Fig. 1 Effects of fesoterodine on vascular voltage-dependent K+ (Kv) currents in coronary arterial smooth muscle cells. Representative traces from vascular smooth muscle cells depolarized from –80 mV to various potentials before (A) and after (B) fesoterodine exposure. (C) Summary of current-voltage (I–V) relationships at steady-state Kv currents in the absence (○) and presence (●) of fesoterodine. n = 8. Only one cell was used from a rabbit to reduce individual differences. *p < 0.05 (control vs. fesoterodine, at each voltage).

• 

  Fig. 2 Concentration dependence of fesoterodine-induced voltage-dependent K+ (Kv) current inhibition. Whole-cell Kv currents were elicited by 600 ms step depolarization to +60 mV from a holding potential of –80 mV. (A) Superimposed traces of Kv currents with increasing concentrations of fesoterodine. (B) Concentration-dependent Kv current inhibition by fesoterodine was measured at a steady-state. The percentage inhibition of Kv currents was plotted against the fesoterodine concentration. The smooth line was fitted to the Hill equation. n = 7.

• 

  Fig. 3 Changes in steady-state activation and inactivation curves by fesoterodine. (A) Activation curves of the voltage-dependent K+ (Kv) currents in the absence (○) and presence (●) of 3 μM fesoterodine. The tail currents were activated by a returning potential of –40 mV after short depolarizing pulses (20–50 ms) at different voltages. n = 7. (B) Inactivation curves of the Kv currents in the absence (○) and presence (●) of 3 μM fesoterodine. The curves were elicited by a test step to +40 mV after 7 s depolarizing pulses at different voltages. n = 6. *p < 0.05 (control vs. fesoterodine, at each voltage).

• 

  Fig. 4 Use (state)-dependence of fesoterodine effects on voltage-dependent K+ (Kv) channels. Twenty repetitive depolarizing one-step pulses from –80 mV to +60 mV were applied at frequencies of 1 Hz (A) and 2 Hz (B) in the absence (○) and presence (●) of 3 μM fesoterodine. Train pulse-induced Kv currents were normalized to the peak current elicited by the first train pulse, and plotted against the pulse number. All n = 6. *p < 0.05 (control vs. fesoterodine, at each pulse number).

• 

  Fig. 5 Effects of fesoterodine on the voltage-dependent K+ (Kv) recovery kinetics from inactivation. The recovery time constant was measured by applying identical twin pulses from –80 mV to +60 mV with extension of intervals from 30 ms to 7 s. Solid lines indicate recovery kinetics from Kv channel inactivation in the absence (○) and presence (●) of 3 μM fesoterodine. n = 5. *p < 0.05 (control vs. fesoterodine at each interval).

• 

  Fig. 6 The role of voltage-dependent K+ (Kv)1.5, Kv2.1, and Kv7 subtypes in the inhibitory effects of fesoterodine. The Kv currents were evoked by 600 ms step depolarization to +60 mV from a holding potential of –80 mV. (A) Superimposed Kv currents under control conditions, in the presence of DPO-1, and fesoterodine + DPO-1. (B) Summary data obtained from (A). n = 4. NS, not significant (DPO-1 vs. fesoterodine + DPO-1). (C) Superimposed Kv currents under control conditions, in the presence of guangxitoxin, and fesoterodine + guangxitoxin. (D) Summary data obtained from (C). n = 5. *p < 0.05 (guangxitoxin vs. fesoterodine + guangxitoxin). (E) Superimposed Kv currents under control conditions, in the presence of linopirdine, and fesoterodine + linopirdine. (F) Summary data obtained from (E). n = 5. *p < 0.05 (linopirdine vs. fesoterodine + linopirdine).

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