The in vitro antibacterial activity of OPC-17116, a new fluoroquinolone, against a wide variety of clinical isolates was evaluated and compared with those of ciprofloxacin, ofloxacin, and norfloxacin. OPC-17116 showed potent broad-spectrum activity against gram-positive and -negative bacteria. The activity of this compound against gram-positive bacteria was higher than those of other quinolones, and its activity against gram-negative and anaerobic bacteria was roughly comparable to those of other quinolones. OPC-17116 had potent activity against important pathogens of respiratory tract infections such as Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Haemophilus influenzae, and Branhamella catarrhalis. The MICs of this compound against 90% of these organisms, except for methicillin-resistant S. aureus, ranged from less than or equal to 0.006 to 3.13 micrograms/ml. OPC-17116 at more than one-half the MICs was bactericidal against clinical isolates of S. aureus, Escherichia coli, K. pneumoniae, and P. aeruginosa. The activity of OPC-17116 was decreased by several culture conditions such as acidic pH, high concentration of Mg2+ ions, and inoculum size of 10(7) CFU/ml. OPC-17116 inhibited the supercoiling activity of DNA gyrases from E. coli KL-16 and S. aureus SA113 (50% inhibitory concentrations, 0.19 and 23.0 micrograms/ml, respectively). The amount of OPC-17116 accumulation was higher than that of other quinolones in S. aureus.