Thiol Reactivity of N-Aryl α-Methylene-γ-lactams: A Reactive Group for Targeted Covalent Inhibitor Design.

Kinase activity can be modulated reversibly or irreversibly by the reaction of targeted covalent inhibitors with nucleophilic residues in protein active sites. Herein, we present thiol reactivity studies that support α-methylene-γ-lactams as tunable surrogates for the highly reactive α-methylene-γ-lactones. The reactivity of the α-methylene is modulated via the N substituent, and the reaction rates toward glutathione were determined via mass spectrometry. Density functional theory calculations of transition states of thiol additions to α-methylene-γ-lactams revealed that the use of the M06-2X functional with the SMD solvation model and methyl thiolate as a model nucleophile reliably predicts the relative reactivities of the α-methylene-γ-lactams, and quasiharmonic approximations improve the agreement between experiment and computation.