2022 - Discovery of novel activators of large-conductance calcium-activated potassium channels for the treatment of cerebellar ataxia

icon sp96 SyncroPatch 384PE (a predecessor model of the SyncroPatch 384 instrument) publication in Molecular Pharmacology (2022)

Authors:
Srinivasan S.R.Huang H.Chang W.C.Nasburg J.A.Nguyen H.M.Strassmaier T.Wulff H., Shakkottai V.G.

Journal:
Molecular Pharmacology (2022) doi:10.1124/molpharm.121.000478


Abstract: 

Impaired cerebellar Purkinje neuron firing resulting from reduced expression of large-conductance calcium-activated potassium (BK) channels is a consistent feature in models of inherited neurodegenerative Spinocerebellar Ataxia (SCA). Restoring BK channel expression improves motor function and delays cerebellar degeneration, indicating that BK channels are an attractive therapeutic target. Current BK channel activators lack specificity and potency and therefore are poor templates for future drug development. We implemented an automated patch-clamp platform for high throughput drug discovery of BK channel activators using the Nanion SyncroPatch 384PE system. We screened over 15,000 compounds for their ability to increase BK channel current amplitude under conditions of lower intracellular calcium that is present in disease. We identified several novel BK channels activators that were then re-tested on the SyncroPatch 384PE to generate concentration-response curves (CRCs). Compounds with favorable CRCs were subsequently tested for their ability to improve irregular cerebellar Purkinje neuron spiking, characteristic of BK channel dysfunction in SCA1 mice. We identified a novel BK channel activator, 4-chloro-N-(5-chloro-2-cyanophenyl)-3-(trifluoromethyl)benzene-1-sulfonamide (herein renamed BK-20), that activated BK currents more potently (pAC50 = 4.64) than NS-1619 (pAC50 = 3.7) at a free internal calcium concentration of 270 nM in a heterologous expression system and improved spiking regularity in SCA1 Purkinje neurons. BK-20 had no activity on SK1-3 channels but interestingly was a potent blocker of Cav3.1 (IC50 = 1.05 mM). Our work describes both a novel compound for further drug development in disorders with irregular Purkinje spiking and a unique platform for drug discovery in degenerative ataxias.

Significance Statement Motor impairment associated with altered Purkinje cell spiking due to dysregulation of BK expression and function is a shared feature of disease in many degenerative ataxias. BK channel activators represent an outstanding therapeutic agent for ataxia. We have developed a high-throughput platform to screen for BK channel activators and identified a novel compound that can serve as a template for future drug-development for the treatment of these disabling disorders.


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