Abstract 5480: Live cell real-time quantification of drug-target engagement for rapid drug discovery

Abstract Drug discovery for challenging drug targets necessitates the proteomic complexities of the cellular milieu for proper folding and function. Therefore, the conventional biophysical methods are not sufficient due to the artificial acellular environment they impose on the target. Cell target engagement is a powerful paradigm in drug discovery, as measures transitions in the thermodynamic state of target protein during engagement with drug molecules. The currently available luminescence-based cell target engagement methods are limited to measurement of end-point signal, poor utility across multiple challenging target families and insufficient resolution and integration for kinetic and mechanistic readouts. We have developed a highly sensitive and versatile fluorescence-based cell target engagement strategy designed for challenging drug targets. The technology enables interrogation of drug targets without interfering with folding, localization and function - all within the physiological milieu of the cell. Importantly, this new method allows for quantitation and monitoring of cell target engagement in real time using live cells. The system seamlessly integrates with readily available real-time systems such as QuantStudio, thus making it highly sensitive and scalable. We share data on challenging drug targets such as Myc, MAPK1, UBE2N and KRAS, and we demonstrate utility of this new methodology for future drug discovery. Citation Format: Ivan Babic, Nikolas Bryan, Claire Cunningham, Avery Sampson, Daniel Starczynowski, Elmar Nurmemmedov. Live cell real-time quantification of drug-target engagement for rapid drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5480.