Development of biophysical methods for characterization of PROTACs

Abstract: Targeting protein with the aim of ubiquitin-labelling for degradation is a new and upand coming field of drug discovery. Proteolysis targeting chimeras (PROTACs) areheterobifunctional linker molecules that connects the target molecule to an E3ubiquitin ligase, subsequent ubiquitination of the target protein initiates the proteindegradation by the proteosome system. The functional nature or mechanism ofPROTACs have the potential to reach previously undruggable proteins with shallowbinding pockets where traditionally designed inhibitors have failed. This projectexamines the possibility to develop a biophysical strategy for characterization andranking of PROTAC compounds. During the project biophysical methods such as SPRwith BIAcore T200, GCI with WAVE, switchSense and MST have been used todevelop a PROTAC ranking assay. Limited solubility of the PROTAC compoundshindered the development of a PROTAC ranking assay, the concentrations neededfor affinity estimation could not be reached due to the solubility of the PROTACcompounds under given condition. Hence further development of the PROTACranking assay is required. There is potentially a great deal of knowledge that can begathered from a working biophysical PROTAC ranking assay that can assist in thedevelopment of new therapeutic compounds.