In conclusion, we report here the use of the pyridine scaffold for the development of inhibitors against VRK1 and VRK2. The most potent compounds 26 and 18 displayed KD values of 190 and 401 nM against VRK1 and VRK2, respectively. Both compounds were not toxic to mammalian cells (up to 3.2 µM). Crystallographic analysis of VRK1 and VRK2 bound to our pyridine analogues highlighted the importance of difluorophenol for binding, and suggested design strategies for the development of specific inhibitors for each of the VRKs. We believe the pyridine scaffold to be a promising core for developing potent and selective inhibitors for VRK1 and VRK2. Future optimization will take advantage of the scaffold low molecular weight and its rapid and efficient two-step synthetic route.