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
It's always satisfying when we are able to apply our expertise to a specific target class resulting in above average results.