Elucidating Compound Selectivity: Integration Of Chemoproteomic, Protein Signaling, And Gene Expression Profiles

Compound profiling technologies that enable an early assessment of cellular effects related to toxicology and pharmacology can contribute to a reduction of late stage failures in drug development. However, the implementation of -omic profiling strategies has been hampered by challenges in analysis and interpretation of these information-rich data sets. To address this issue, we have rationally combined analytical methods that report compound-protein interactions (Inhibitor Affinity Capture/IAC; protein arrays) with technologies that monitor down-stream cellular effects on protein signaling (Protein-fragment Complementation Assays) and gene expression. IAC is a mass spectrometry-based technique that discriminates tight from weaker binding proteins using a label-free affinity binning process. Using these orthogonal approaches and pathway analysis tools, we have: 1) provided additional comprehension of mechanisms driving relevant cellular endpoints for marketed oncology drugs; 2) revealed off-target profiles for several experimental kinase inhibitors; 3) further elucidated mechanisms of toxicity using a series of fluoroquinolone antibiotics. The multiple lines of evidence generated from this integrated technology scheme provide higher confidence decision-making and improves the quality of compounds that are progressed towards the clinic.