Differential efficacy of GoSlo-SR compounds on BKα and BKαγ(1-4) channels

GoSlo-SR化合物对BKα和BKαγ(1-4)通道的差异性作用

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Abstract

Large conductance, voltage and Ca(2+) activated K(+) channels (BK channels) are abundantly expressed throughout the body and are important regulators of smooth muscle tone and neuronal excitability. Their dysfunction is implicated in various diseases including overactive bladder, hypertension and erectile dysfunction. Therefore, BK channel openers bear significant therapeutic potential to treat the above diseases. GoSlo-SR compounds were designed to be potent and efficacious BK channel openers. Although their structural activity relationships, activation in both BKα and BKαβ channels and the hypothetical mode of action of these compounds has been studied in detail in recent years, their effectiveness to open the BKαγ channels still remains unexplored. In this study, we have examined the efficacy of 3 closely related GoSlo-SR openers, GoSlo-SR-5-6 (SR-5-6), GoSlo-SR-5-44 (SR-5-44) and GoSlo-SR-5-130 (SR-5-130) using inside out patches on BKα channels coexpressed with 4 different LRRC (γ(1-4)) subunits in HEK293 cells. Our data suggests that the activation effects due to SR-5-6 were not significantly affected in the presence of γ(1-4) subunits. Interestingly, the effects of more efficacious BK channel opener SR-5-44 were altered by different γ subunits. In cells expressing BKα channels, the shift in V(1/2) (ΔV(1/2)) induced by SR-5-44 (3 μM) was -76 ± 3 mV, whereas it was significantly reduced by ∼70 % in BKαγ(1) channels (ΔV(1/2)= -23 ± 3, p < 0.001, ANOVA). In BKαγ(2) channels the ΔV(1/2) was -36 ± 1 mV, which was less than that observed in BKαγ(3) and BKαγ(4) channels where the ΔV(1/2) was -47 ± 5 mV, and -82 ± 5 mV, respectively. Additionally, the excitatory effects of a 'β specific' BK channel opener, SR-5-130 were only partially restored in the patches containing BKαγ(1-4) channels. Together this data highlights that subtle modifications in GoSlo-SR structures alter their effectiveness on BK channels with accessory γ subunits and this study might provide a scaffold for the development of more tissue specific BK channel openers.

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