High-Throughput Screening Of Nav1.7 Sodium Channel Blockers In An Automated Patch Clamp Assay

The voltage gated sodium channel Nav1.7 is a well validated target for pain in humans. Among the many technology platforms used to identify modulators of Nav1.7, patch clamp electrophysiology remains the gold standard, however its utility in drug discovery is historically limited by its low throughput. In the past decade, automated patch clamp (APC) technology has undergone a major revolution, closing the gap of utilizing electrophysiology for ion channel screening. In this study, we implemented the latest generation of APC platform, SyncroPatch 768PE, which can perform parallel recording in dual 384 modules with giga-seal level data quality. By optimizing various parameters (cell catching, signal window and signal stability, etc.), we developed high quality Nav1.7 assay with success rate of 81%, Z’ factor of 0.72; and correlation coefficients R u003e 0.9 with manual patch clamp electrophysiology. We also implemented a GeneData analysis tool to handle the large amount of data. In a pilot study, we screened ∼10,000 compounds within 5 days, and conducted hit confirmation, subtype selectivity and mechanism of action studies using the same APC platform. Therefore the SyncroPatch as well as new high throughput data analysis tools represent a significant advance in Nav1.7 modulator screening and lead profiling.