Inhibition of tolbutamide 4-methylhydroxylation by a series of non-steroidal anti-inflammatory drugs in V79-NH cells expressing human cytochrome P4502C10.
1. To study the role of cytochrome P4502C10 in the metabolism of the non-steroidal antiinflammatory drugs (NSAIDs) diclofenac, phenylbutazone, fenoprofen, ibuprofen, flurbiprofen, ketoprofen and naproxen, a cell line was developed stably expressing CYP2C10 cDNA. A retroviral vector construct, containing a human CYP2C10 cDNA, was transfected in V79-NH Chinese hamster lung cells by calcium phosphate co-precipitation. Sublines stably expressing human cytochrome P450 cDNA were established by selection with the neomycin analogue G418. 2. Enzymatic activity of CYP2C10 was detected by 4-methylhydroxylation of tolbutamide. This activity was inhibited to background levels by preincubation with the CYP2C9/10 inhibitor sulphaphenazole. 3. Preincubations with the NSAIDs ketoprofen, phenylbutazone, flurbiprofen and diclofenac (all 250 microM) caused a decrease in 4-methylhydroxylation of tolbutamide (500 microM), significantly different from control values (p < 0.05). Inhibition of this activity was not seen in preincubations with the NSAIDs fenoprofen, ibuprofen and naproxen (250 microM). 4. The V79-NH CYP2C10 cell line we have developed has been shown to be a useful tool to predict drug-drug interactions.
Kappers
W A
WA
Research Institute of Toxicology, Utrecht University, The Netherlands.
Groene
E M
EM
Kleij
L A
LA
Witkamp
R F
RF
Zweers-Zeilmaker
W M
WM
Feron
V J
VJ
Horbach
G J
GJ
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D001189:Aryl Hydrocarbon Hydroxylases; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D065729:Cytochrome P-450 CYP2C9; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004008:Diclofenac / Q000494:pharmacology; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005480:Flurbiprofen / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D006900:Hydroxylation; D007660:Ketoprofen / Q000494:pharmacology; D008168:Lung; D008745:Methylation; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D010653:Phenylbutazone / Q000494:pharmacology; D013426:Sulfaphenazole / Q000494:pharmacology; D014044:Tolbutamide / Q000378:metabolism*; D014162:Transfection
D000894:Anti-Inflammatory Agents, Non-Steroidal; D004791:Enzyme Inhibitors; D013426:Sulfaphenazole; D004008:Diclofenac; D005480:Flurbiprofen; D003577:Cytochrome P-450 Enzyme System; D007660:Ketoprofen; D014044:Tolbutamide; D006899:Mixed Function Oxygenases; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases; C053553:tolbutamide 4-hydroxylase; D010653:Phenylbutazone