K-2P (KCNK) potassium channels create "leak" potassium currents that strongly influence cellular excitability and contribute to discomfort, somatosensation, anesthesia, and mood. Despite their physiological value, K(2P)s lack precise pharmacology. Addressing not this concern continues to be complex through the difficulties the leak nature of K-2P currents poses for electrophysiology-based high-throughput screening strategies. Here, we present a yeast-based high-throughput screening assay that avoids this difficulty. Using a simple growth-based practical readout, we screened a library of 106,281 tiny molecules and identified two new inhibitors and three new activators on the mammalian K-2P channel K(2P)two.1 (KCNK2, TREK-1).
By combining biophysical, structure-activity, and mechanistic evaluation, we formulated a dihydroacridineLinifanib (ABT-869) analogue, ML67-33, that acts as a very low micromolar, selective activator of temperature- and mechano-sensitive K-2P channels. Biophysical studies display that ML67-33 reversibly increases channel currents by activating the extracellular selectivity filter-based C-type gate that kinds the core gating apparatus on which several different varied modulatory inputs converge. The new K-2P modulators presented here, together with the yeast-based assay, really should enable each mechanistic and physiological scientific studies of K-2P activity and facilitate the discovery and development of other K-2P compact molecule selleck chem inhibitor modulators.