Chemical and Computational Methods for the Characterization of Covalent Reactive Groups for the Prospective Design of Irreversible Inhibitors

• DOI: 10.1021/jm501412a
• Publication Date: 2014-11-06T16:42:43Z
• Authors: Mark E. Flanagan; Joseph A. Abramite; Dennis P. Anderson; Ann Aulabaugh; Upendra P. Dahal; Adam M. Gilbert; Chao Li; Justin Montgomery; Stacey R. Oppenhe...; Tim Ryder; Brandon P. Schuff; Daniel P. Uccello; Gregory S. Walker; Yan Wu; Matthew F. Brown; Jinshan M. Chen; Matthew M. Hayward; Mark C. Noe; R. Scott Obach; Laurence Philippe; Veerabahu Shanmug...; Michael J. Shapiro; Jeremy Starr; Justin Stroh; Ye Che
• Keywords: 0301 basic medicine; Kinetics; 03 medical and health sciences; Pharmaceutical Preparations; Drug Design; Humans; Enzyme Inhibitors; Glutathione; Nuclear Magnetic Resonance, Biomolecular; Mass Spectrometry

Interest in drugs that covalently modify their target is driven by the desire for enhanced efficacy that can result from the silencing of enzymatic activity until protein resynthesis can occur, along with the potential for increased selectivity by targeting uniquely positioned nucleophilic residues in the protein. However, covalent approaches carry additional risk for toxicities or hypersensitivity reactions that can result from covalent modification of unintended targets. Here we describe methods for measuring the reactivity of covalent reactive groups (CRGs) with a biologically relevant nucleophile, glutathione (GSH), along with kinetic data for a broad array of electrophiles. We also describe a computational method for predicting electrophilic reactivity, which taken together can be applied to the prospective design of thiol-reactive covalent inhibitors.