Probing the KRas Switch II Groove by Fluorine NMR Spectroscopy

Abstract

While there has been recent success in the development of KRas^G12C inhibitors, unmet needs for selective inhibitors of KRas^G12D and the remaining oncogenic KRas proteins remain. Here, we applied trifluoromethyl-containing ligands of KRas proteins as competitive probe ligands to assay the occupancy of the switch II pocket by ¹⁹F NMR spectroscopy. Structure-activity-relationship studies of probe ligands increased the sensitivity of the assay and identified structures that differentially detected each nucleotide state of KRas^G12D. These differences in selectivity, combined with the high resolution of ¹⁹F NMR spectroscopy, enabled this method to be expanded to assay both nucleotide states of the protein simultaneously.