Inhibition of tolbutamide 4-methylhydroxylation by a series of non-steroidal anti-inflammatory drugs in V79-NH cells expressing human cytochrome P4502C10
article
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 coprecipitation. 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 μM) caused a decrease in 4-methylhydroxylation of tolbutamide (500 μM), 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 αM).
4. The V79-NH CYP2C10 cell line we have developed has been shown to be a useful tool to predict drug-drug interactions.
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 μM) caused a decrease in 4-methylhydroxylation of tolbutamide (500 μM), 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 αM).
4. The V79-NH CYP2C10 cell line we have developed has been shown to be a useful tool to predict drug-drug interactions.
Topics
CYP2C9 protein, humanCytochrome P450DiclofenacEnzyme inhibitorFlurbiprofenKetoprofenMixed function oxidaseNonsteroid antiinflammatory agentPhenylbutazoneSulfaphenazoleTolbutamideTolbutamide 4 hydroxylaseTolbutamide 4-hydroxylaseUnspecific monooxygenaseAnimalCell lineCricetulusDrug interactionGene expressionGenetic transfectionGeneticsHamsterHumanHydroxylationLungMetabolismMethylationAnimalsAnti-Inflammatory Agents, Non-SteroidalAryl Hydrocarbon HydroxylasesCell LineCricetinaeCricetulusCytochrome P-450 Enzyme SystemDiclofenacDrug InteractionsEnzyme InhibitorsFlurbiprofenGene ExpressionHumansHydroxylationKetoprofenLungMethylationMixed Function OxygenasesPhenylbutazoneSulfaphenazoleTolbutamideTransfection
TNO Identifier
233653
ISSN
00498254
Source
Xenobiotica, 26(12), pp. 1231-1239.
Pages
1231-1239
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