Inhibition of hERG by siRNA Takes More Than 8 hrs to Manifest: Implications for Designing hERG Assays

An integrated assay platform was developed to characterise the pharmacological and physiological effects of Kv7.2/7.3 activators. Combining high-throughput FLIPR thallium flux, automated voltage- and current-clamp electrophysiology, and sensory neuron recordings enabled assessment of compound potency, mechanism of action, and effects on neuronal resting membrane potential. Profiling of retigabine, flupirtine, ML213, ICA 069673, XEN1101 and opakalim demonstrated distinct pharmacological profiles across complementary assay systems. The workflow provides a mechanistic and translational approach for characterising Kv7.2/7.3 activators and their effects on neuronal excitability.

Optical voltage imaging of human iPSC-derived cardiomyocytes was used to assess electrophysiological effects of compounds beyond hERG inhibition. Action potential waveform analysis revealed compound-specific and concentration-dependent changes, enabling mechanistic differentiation of multichannel activity and demonstrating a human-relevant approach for translational cardiac safety assessment.