In the development of PROTACs it is vital to have a varied suite of assays that can inform the structure-activity relationship (SAR) and progress the design-make-test cycle. A number of PROTACs have previously been developed to target the well-studied protein BRD4 – a member of the BET (bromodomain and extra-terminal) family of proteins whose dysregulation is linked to human cancers.
At Charnwood Discovery we have used cell-based and biophysical techniques to investigate – amongst others – the BRD4-targeting PROTAC ‘dBET6’ to elucidate a binding mechanism, confirm proteasomal degradation and monitor its impact on cellular behavior. Here we demonstrate the techniques that we might typically implement in a PROTAC development cascade.
