Intermolecular interactions of antimicrobial fluoroquinolones with purified rat liver CYP1A2 studied by proton nuclear magnetic resonance spectroscopy.
1. Binding and inhibition of antimicrobial fluoroquinolones towards liver CYP1A2 purified from 3-methylcholanthrene-treated rats were investigated using proton nuclear magnetic resonance (nmr) and phenacetin metabolism. 2. The proton nmr longitudinal relaxation rate study indicated that the paramagnetic effects of the haem iron of CYP1A2 were observed in protons of enoxacin with a 1,8-naphthyridine skeleton and its 4'-nitrogen atom on the 7-piperazine ring probably participated in specific binding to the haem iron. These data suggest a facile accessibility and strong binding of enoxacin to the active site of the enzyme. On the contrary, the binding region of norfloxacin with a quinoline skeleton could not be specified, and an 8-fluorinated derivative (AT-3970) had much lower paramagnetic effects and no specific binding region. 3. In a reconstituted CYP1A2 system, enoxacin exhibited the most potent inhibition of phenacetin O-deethylation. The metabolism was less inhibited by norfloxacin, and AT-3970 had a weak inhibitory activity. 4. The binding ability of the fluoroquinolones to the CYP1A2 active site is likely to determine their inhibitory activity against phenacetin metabolism.
Mizuki
Y
Y
Developmental Research Laboratories, Dainippon Pharmaceutical Co. Ltd., Osaka, Japan.
Yamamoto
K
K
Yamaguchi
T
T
Fujii
T
T
Miyazaki
H
H
Ohmori
H
H
D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism*; D002939:Ciprofloxacin / Q000031:analogs & derivatives / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000302:isolation & purification / Q000378:metabolism*; D015365:Enoxacin / Q000378:metabolism; D024841:Fluoroquinolones; D008099:Liver / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009643:Norfloxacin / Q000378:metabolism; D010615:Phenacetin / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D000890:Anti-Infective Agents; D024841:Fluoroquinolones; D015365:Enoxacin; D002939:Ciprofloxacin; C103419:8-fluorociprofloxacin; D019388:Cytochrome P-450 CYP1A2; D010615:Phenacetin; D009643:Norfloxacin