Target deconvolution of small molecules is critical to understanding a potential therapeutics mechanism of action and safety liabilities. Although affinity pull down approaches have greatly expanded our knowledge of drug targets, derivatization of the drug requires domain expertise in medicinal chemistry. Furthermore, affinity pull downs are not feasible when a drug molecule has a steep SAR that precluded linker attachment or lack of chemistry resources. Cellular thermal shift assay coupled with high-resolution mass-spectrometry (CETSA®-MS) has shown to be indispensable method for unbiased, proteome-wide, chemistry free target deconvolution by measuring changes in protein thermal stability upon compound binding within physiologically relevant system. The method is increasingly being employed both in mechanism of action (MoA) studies and to identify primary and off-targets of candidate drug molecules.
In collaboration with Pelago, AstraZeneca have applied cCETSA (compressed CETSA), a more time and cost effective format of next generation CETSA MS on drug discovery projects. Here we would like to present the application of cCETSA in AstraZeneca for compounds at various project stages like phenotypic HTS hit, candidate identification drug, lead optimisation identification drug, literature compounds there by influencing key project decisions.
