Lansoprazole enantiomer activates human liver microsomal CYP2C9 catalytic activity in a stereospecific and substrate-specific manner.
We recently proposed a possible stereoselective activation by lansoprazole of CYP2C9-catalyzed tolbutamide hydroxylation, as well as stereoselective inhibition of several cytochrome P450 (P450) isoforms. This study evaluated the effects of lansoprazole enantiomers on CYP2C9 activity in vitro, using several probe substrates. For tolbutamide 4-methylhydroxylation and phenytoin 4-hydroxylation, R-lansoprazole was an activator (140 and 550% of control at 100 microM R-lansoprazole, EC50 values of 19.9 and 30.2 microM, respectively). R-Lansoprazole-mediated activation of the formation of 4-hydroxyphenytoin was also seen with recombinant human CYP2C9. R-Lansoprazole increased the Michaelis-Menten-derived V(max) of phenytoin 4-hydroxylation from 0.024 to 0.121 pmol/min/pmol P450, and lowered its K(m) from 20.5 to 15.0 microM, suggesting that R-lansoprazole activates CYP2C9-mediated phenytoin metabolism without displacing phenytoin from the active site. Kinetic parameters were also estimated using the two-site binding equation, with alpha values <1 and beta values >1, indicative of activation. Additionally, phenytoin at 10 to 200 microM had no reciprocal effect on the hydroxylation of R-lansoprazole. Meanwhile, R-lansoprazole had no activation effect on diclofenac and S-warfarin metabolism in the incubation study using both recombinant CYP2C9 and human liver microsomes. These substrate-dependent activation effects suggest that phenytoin has a different binding orientation compared with diclofenac and S-warfarin. Overall, these results suggest that R-lansoprazole activates CYP2C9 in a stereospecific and substrate-specific manner, possibly by binding within the active site and inducing positive cooperativity. This is the first report to describe stereoselective activation of this cytochrome P450 isoform.
Liu
Kwang-Hyeon
KH
Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine and Clinical Pharmacology Center, Busan Paik Hospital, Korea.
Kim
Min-Jung
MJ
Jung
Woo Moon
WM
Kang
Wonku
W
Cha
In-June
IJ
Shin
Jae-Gook
JG
D053799:2-Pyridinylmethylsulfinylbenzimidazoles; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002384:Catalysis / Q000187:drug effects; D065729:Cytochrome P-450 CYP2C9; D064748:Dexlansoprazole; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006801:Humans; D064747:Lansoprazole; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology; D009853:Omeprazole / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000494:pharmacology*; D013237:Stereoisomerism; D013379:Substrate Specificity / Q000187:drug effects / Q000502:physiology
D053799:2-Pyridinylmethylsulfinylbenzimidazoles; D064747:Lansoprazole; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; D009853:Omeprazole; D064748:Dexlansoprazole