Novel approach for comparing the abilities of quinolones to restrict the emergence of resistant mutants during quinolone exposure.
An agar-plate assay was adapted to examine aspects of quinolone structure that restrict the emergence of quinolone-mediated quinolone resistance. When Escherichia coli was applied to agar containing nalidixic acid, the number of quinolone-resistant mutants arising during incubation was decreased by raising the drug concentration and by mutations expected to block the induction of the SOS response (recA, lexA); the mutant number was increased by a mutator mutation (ung). The examination of four related fluoroquinolones then revealed that a C-8 methoxy group and an N-ethyl piperazine substituent at C-7 reduced mutant acquisition more effectively than C-8 H and C-7 C-ethyl piperazine groups. The fluoroquinolone that was most effective at restricting mutant acquisition was the most active when lethal activity was measured on agar plates or in liquid medium (as minimal bactericidal concentration). It also exhibited the lowest ratio of mutant MIC to wild-type MIC when it was tested with a set of isogenic gyrase mutants, and it had a low mutant prevention concentration (MPC) relative to MIC. However, a low MPC was less likely to be important in restricting the induced mutant accumulation because a fluoroquinolone N-ethyl piperazine substituent was more effective than a C-ethyl piperazine substituent at reducing mutant accumulation but was less effective at lowering the MPC. An 8-methoxy-quinazoline-2,4-dione was also effective at restricting the accumulation of resistant mutants on agar. Collectively, these data characterize a simple assay for detection of drug-mediated resistance that is sensitive to the structures of GyrA inhibitors. The assay provides a new method for screening quinolones and quinolone-like molecules that complements MPC-based tests for restricting the emergence of resistance.
Malik
Muhammad
M
Public Health Research Institute, New Jersey Medical School, UMDNJ, 225 Warren Street, Newark, NJ 07103, USA.
Hoatam
Gerard
G
Chavda
Kalyan
K
Kerns
Robert J
RJ
Drlica
Karl
K
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D016587:Antimutagenic Agents / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects / Q000235:genetics*; D001483:Base Sequence; D003470:Culture Media; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D024841:Fluoroquinolones / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009268:Nalidixic Acid / Q000494:pharmacology; D059005:Topoisomerase II Inhibitors
D000900:Anti-Bacterial Agents; D016587:Antimutagenic Agents; D003470:Culture Media; D024841:Fluoroquinolones; D059005:Topoisomerase II Inhibitors; D009268:Nalidixic Acid