Predicted multiple selected reaction monitoring to screen activated drug-mediated modifications on human serum albumin

摘要: Abstract Metabolic activation of drugs frequently generates electrophilic products that may undergo covalent binding to biological macromolecules, such as proteins and DNA. The resulting adducts are considerable concern in drug discovery development. Several strategies for assessing the potential risks candidate have been reported. Of these, glutathione trapping is most commonly used method together with mass spectrometry. Furthermore, drug-mediated protein modifications studied using serum albumin CYP enzymes clarify target amino acids mechanism-based inhibition, respectively. In this article, we introduce a practical way screen modifications. method, referred “predicted multiple selected reaction monitoring,” based on monitoring (SRM) strategy, but targets all possible chemically modified tryptic peptides. creation SRM lists require patience; however, strategy could facilitate more sensitive screening compared common data-dependent product ion scanning. Ketoprofen- N -hydroxysuccinimidyl ester (equivalent glucuronide) -acetyl- p -benzoquinone imine (NAPQI) were allowed react human model experiment. Using 11 ketoprofen-adduction sites (at Lys 137, 195, 199, 212, 351, 402, 432, 436, 525, 536, 541 ) 1 NAPQI-adduction site Cys 34 easily identified.