Inhibitory effective perturbations of cilobradine (DK-AH269), a blocker of HCN channels, on the amplitude and gating of both hyperpolarization-activated cation and delayed-rectifier potassium currents

Cilobradine (CIL, DK-AH269), an inhibitor of hyperpolarization-activated cation current (I_h), has been observed to possess pro-arrhythmic properties. Whether and how CIL is capable of perturbing different types of membrane ionic currents existing in electrically excitable cells, however, is incompletely understood. In this study, we intended to examine possible modifications by it or other structurally similar compounds of ionic currents in pituitary tumor (GH₃) cells and in heart-derived H9c2 cells. The standard whole-cell voltage-clamp technique was performed to examine the effect of CIL on ionic currents. GH₃-cell exposure to CIL suppressed the density of hyperpolarization-evoked I_h in a concentration-dependent manner with an effective IC₅₀ of 3.38 µM. Apart from its increase in the activation time constant of I_h during long-lasting hyperpolarization, the presence of CIL (3 µM) distinctly shifted the steady-state activation curve of I_h triggered by a 2-s conditioning pulse to a hyperpolarizing direction by 10 mV. As the impedance-frequency relation of I_h was studied, its presence raised the impedance magnitude at the resonance frequency induced by chirp voltage. CIL also suppressed delayed-rectifier K⁺ current (I_K(DR)) followed by the accelerated inactivation time course of this current, with effective IC₅₀ (measured at late I_K(DR)) or K_D value of 3.54 or 3.77 µM, respectively. As the CIL concentration increased 1 to 3 µM, the inactivation curve of I_K(DR) elicited by 1-or 10-s conditioning pulses was shifted to a hyperpolarizing potential by approximately 10 mV, and the recovery of I_K(DR) inactivation during its presence was prolonged. The peak Na⁺ current (I_Na) during brief depolarization was resistant to being sensitive to the presence of CIL, yet to be either decreased by subsequent addition of A-803467 or enhanced by that of tefluthrin. In cardiac H9c2 cells, unlike the CIL effect, the addition of either ivabradine or zatebradine mildly led to a lowering in I_K(DR) amplitude with no conceivable change in the inactivation time course of the current. Taken together, the compound like CIL, which was tailored to block hyperpolarization-activated cation (HCN) channels effectively, was also capable of altering the amplitude and gating of I_K(DR), thereby influencing the functional activities of electrically excitable cells, such as GH₃ cells.