Comprehensive Approach of F-19 Nuclear Magnetic Resonance, Enzymatic, and In Silico Methods for Site-Specific Hit Selection and Validation of Fragment Molecules that Inhibit Methionine gamma-Lyase Activity

摘要

Fragment-based screening using F-19 NMR (F-19-FS) is an efficient method for exploring seed and lead compounds for drug discovery. Here, we demonstrate the utility and merits of using F-19-FS for methionine gamma-lyase-binding fragments, together with a F-19 NMR-based competition and mutation assay, as well as enzymatic and in silico methods. F-19 NMR-based assays provided useful information on binding between F-19-FS hit fragments and target proteins. Although the F-19-FS and enzymatic assay were weakly correlated, they show that the F-19-FS hit fragments contained compounds with inhibitory activity. Furthermore, we found that in silico calculations partially account for the differences in activity levels between the F-19-FS hits as per NMR analysis. A comprehensive approach combining the F-19-FS and other methods not only identified fragment hits but also distinguished structural differences in chemical groups with diverse activity levels.