Study of coumarin metabolism by Chinese hamster lung fibroblasts expressing a human cytochrome P450 using H-nmr.
1. Human cytochrome P450 2A6 is expressed in Chinese hamster lung fibroblasts. The isozyme hydroxylates coumarin at the 7-position. 2. Metabolism of coumarin in these lung fibroblasts was monitored using 1H-nmr. Media samples were taken from cells grown in flasks and also in a fluidized bed bioreactor. In each case 7-hydroxycoumarin was readily observable by nmr in crude extracts of the medium. 3. The rate of formation of 7-hydroxycoumarin observed in the acute bioreactor experiments on a per cell basis was found to be much higher than that obtained under chronic monolayer conditions, and correlated with glucose consumption rates.
Street
J C
JC
Departments of Central Technologies and Drug Safety and Metabolism, Sandoz Research Institute, East Hanover, NJ 07936, USA.
Lee
J S
JS
Jarema
M A
MA
D000818:Animals; D002460:Cell Line; D003374:Coumarins / Q000378:metabolism* / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D014468:Umbelliferones / Q000378:metabolism; D015262:Xenobiotics / Q000378:metabolism
D003374:Coumarins; D003470:Culture Media; D014468:Umbelliferones; D015262:Xenobiotics; C031477:7-hydroxycoumarin; D003577:Cytochrome P-450 Enzyme System; C030123:coumarin