Ribocil-C is a potent inhibitor of bacterial riboflavin riboswitches. Riboswitches are cis regulatory elements present in non-coding RNA th at specifically bind to natural ligands to regulate gene expression. Ribocil-C blocks the flavin mononucleotide riboswitch-mediated expression of the ribB gene, which is required for riboflavin biosynthesis, with an IC50 value of 23 nM. Ribocil-C inhibited bacterial cell growth of E. coli MB5746 and bacterial burden in a mouse E. coli septicaemia model of infection.