Benzodioxane-benzamide FtsZ inhibitors: Synthesis of new derivatives and their biophysical and biochemical evaluation

The bacterial cell division is a complex process, leading to the scission of the parent cell in two daughter cells, genetically identical and having similar size. The achievement of a correct division is regulated by several bacterial proteins. This fine control involves the correct localization of the division septum as well as the ring constriction, that finally allows the physical separation. Among these crucial proteins, FtsZ has been universally recognized as the main actor and, consequently, as a potential and useful target for the obtainment of new antimicrobials. Indeed, FtsZ polymerizes at the centre of the cell into a circular structure (named Z-ring) which recruits all the downstream proteins that lead to cell division. In the last years, we developed a huge series of benzamide inhibitors having high antimicrobial activity, which proved to be related to the inhibition of FtsZ. In this poster, we present the synthesis of the recent derivatives, together with the results of the biophysical and biochemical characterization of the interaction with FtsZ. In particular, we determined FtsZ polymerization/depolymerization properties by fluorescence anisotropy and we estimated FtsZ polymers size by analytical ultracentrifugation. We also gathered information, using microscopy techniques, on how our compounds influence the morphology of the FtsZ polymers. In particular, we studied FtsZ polymerization in dilute solution and in the presence of macromolecular crowding agents, which mimic the crowded environment of the cytoplasm. These analyses showed a correlation between the in vivo potency of the compounds and the level of in vitro FtsZ inhibition, allowing us to validate the target and to trace the path to future investigations.