Pyrimidin-2(1H)-ones based inhibitors of Mycobacterium tuberculosis orotate phosphoribosyltransferase

► Selection and optimization of inhibitors based on MtOPRT kinetic mechanism. ► Identification of MtOPRT inhibitors with nanomolar inhibition constant values. ► In vitro kinetic assays to characterize binding mode of inhibitors. ► Thermodynamic discrimination profiles of inhibitors assessed by ITC. ► Inhibitory activity of 13 is likely due to entropy optimization.