NBTI 5463 is a bacterial type II topoisomerase (topoisomerase II) inhibitor with antimicrobial properties. It effectively inhibits GyrA and TopoIV in Pseudomonas aeruginosa and Escherichia coli. By binding to topoisomerase II, NBTI 5463 prevents DNA cleavage and re-ligation, thereby impeding bacterial DNA replication and transcription. This compound shows potential for research into Gram-negative bacterial infections.