Antibiotic resistance has grown significantly in the last three decades, while research and development of new antibiotic classes has languished. Therefore, new chemical frameworks for the control of microbial behavior are urgently required. This study presents a novel suite of compounds, based on a tricyclic 4-hydroxy-2 H -pyrano[3,2- c ]quinoline-2,5(6 H )-dione core, with significant antibiotic activity against the ESKAPE pathogens Staphylococcus aureus and Enterococcus faecalis and the "accidental pathogen" Staphylococcus epidermidis . A potent analogue with an N -heptyl-9- t- Bu substitution pattern emerged as a hit with MIC levels ≤2 μg/mL across four strains of MRSA. In addition, the same compound proved highly potent against Enterococcus spp. (0.25 μg/mL)., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)