We worked with our client who had no prior knowledge of chemistry and required HTS of the target equivalent to 50,000 compounds. A hit identification rate of 65% with high reproducibility was achieved.

• Target = TRESK – 2 pore non-voltage gated potassium channel
• Disease indication = Migraine (prevalent, debilitating and costly)
• The potassium channel TRESK is abundant in trigeminal ganglia. The mutation causes loss of function, thereby increasing TG excitability. TRESK is therefore a viable therapeutic target for migraine
• TRESK is activated strongly by second messengers such as calclum and volatile anaesthetics such as halothane
• At resting membrane potential, channel is closed but activated by calcium. As a membrane depolarizes, potassium outflow through open TRESK channels tends to restore the membrane potential
• Compound targeting TRESK and to increase TRESK opening in the resting state would reduce excitability and responsiveness
  Case Study – Hit Identification
• Target was transiently expressed in U2OS cells using a Bacmam construct
• FluxOR™ Potassium Ion Channel Assay in FLIPR Tetra
• Selectivity required against TREK channel (heart) therefore selectivity assay developed
• Almost all hits are TRESK specific