title: Quinolones with enhanced bactericidal activity induce autolysis in Streptococcus pneumoniae.
abstract: OBJECTIVE
  DC-159a and sitafloxacin show greater bactericidal activity against Streptococcus pneumoniae than garenoxacin and other quinolones. We investigated whether the autolysis induced by these quinolones contributes to their rapid bactericidal activity.
  
  
  METHODS
  Time-kill studies were conducted against a S. pneumoniae clinical isolate in broth with choline chloride, which is known to inhibit autolytic amidases, and lytA mutants. Western blot analysis was performed to examine LytA production. Scanning electron microscopy (SEM) was used to investigate morphological differences after exposure to quinolone.
  
  
  RESULTS
  Bactericidal activity of DC-159a and sitafloxacin against S. pneumoniae at 2 h of exposure to twice the minimum inhibitory concentration (MIC) was found to decrease by approximately 1 log CFU/ml when autolytic amidases were blocked. Time-kill studies using lytA mutants showed that DC-159a exhibited slower killing than that against the lytA-positive strains. On exposure to the MIC and twice the MIC of DC-159a and sitafloxacin, R6 and a clinical isolate overexpressed LytA, while garenoxacin caused a less significant increase in LytA than DC-159a and sitafloxacin. Scanning electron microscopy images revealed that R6 treated with DC-159a underwent distinct morphological changes, while the lytA mutant did not.
  
  
  CONCLUSIONS
  The present study demonstrated that quinolone-induced autolysis may provide quinolones more powerful bactericidal activity against S. pneumoniae.
authors.affiliation: Biological Research Laboratories IV, Daiichi Sankyo Co Ltd, Tokyo, Japan. okumura.ryo.x7@daiichisankyo.co.jp
authors.forename: Ryo
authors.initials: R
authors.lastname: Okumura
authors.forename: Kazuki
authors.initials: K
authors.lastname: Hoshino
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Otani
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamamoto
mesh_terms: D000631:Aminopyridines / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001433:Bacteriolysis / Q000187:drug effects*; D024841:Fluoroquinolones / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests / Q000379:methods; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism / Q000502:physiology; D013296:Streptococcus pneumoniae / Q000187:drug effects* / Q000235:genetics / Q000648:ultrastructure
chemical_list: C530475:7-(7-amino-7-methyl-5-azaspiro(2.4)heptan-5-yl)-6-fluoro-1-(2-fluoro-1-cyclopropyl)-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid; D000631:Aminopyridines; D000900:Anti-Bacterial Agents; D024841:Fluoroquinolones; C076246:sitafloxacin; D009238:N-Acetylmuramoyl-L-alanine Amidase; C419689:garenoxacin