Objective

Abstract text: Targeted therapies are dependent on adequate occupancy and target engagement to reach their efficacy measure. Yet, more than half of novel candidate drugs entering clinical trial stage fail before proof of concept is achieved. Inadequate target engagement is often at the root cause of these failures. Therefore we have developed a versatile label-free method to quantify the target engagement of drugs on their native physiological relevant targets. The method is validated in multiple sample matrices from bench to bedside: in living cells and tissue from rodents and man. CETSA assays enable translation of compound target engagement between different sample matrices. The relative target engagement potency of lead compounds can be compared and the most promising further profiled in vivo. The rapid assay validation and label free format allows a physiological relevant translation from in vitro to in vivo, devoid of systematic errors from use of different methods. Therefore the use of CETSA in lead profiling will shorten development time and enable selection of candidate drugs with higher likelihood of efficacy. Authors: Jakob Karen (1), Joakim Hellner (1), Nancy Dekki (1), Isabel Caballero (1), Jelena Almqvist (2), Hanna Axelsson (2), Smaranda Bacanu (3), Sara Lööf (3), Takahiro Seki (3), Yihai Cao (3), Pär Nordlund (3) , Daniel Martinez Molina (1) 1≤ Pelago Bioscience, Stockholm Sweden 2≤ Chemical Biology Consortium, Stockholm Sweden 3≤ Karolinska Institutet , Stockholm Sweden