The guinea-pig expresses functional CYP2C and P-glycoprotein: further validation of its usefulness in drug biotransformation/transport studies

被引：1
作者：
Hasibu, Ibrahim [1 ,2 ]
Patoine, Dany [1 ,2 ]
Pilote, Sylvie [1 ,2 ]
Drolet, Benoit [1 ,2 ]
Simard, Chantale [1 ,2 ]
机构：
[1] Univ Laval, Fac Pharm, Quebec City, PQ, Canada
[2] Inst Univ Cardiol & Pneumol Quebec, Quebec City, PQ G1V 4G5, Canada
来源：
BIOPHARMACEUTICS & DRUG DISPOSITION | 2015年 / 36卷 / 03期
基金：
加拿大健康研究院;
关键词：
guinea-pig; CYP2C; P-glycoprotein; drugs; MECHANISM-BASED INACTIVATION; SULFAPHENAZOLE DERIVATIVES; CYTOCHROME P4502C9; EPOXIDE HYDROLASE; METABOLISM; DICLOFENAC; RECOGNITION; ENZYMES; INHIBITION; CLONING;
D O I：
10.1002/bdd.1931
中图分类号：
R9 [药学];
学科分类号：
1007 ;
摘要：
Background: The guinea-pig is an excellent animal model for studying cardiopulmonary physiology/pharmacology. Interestingly, it also possesses a number of drug-metabolizing enzymes found in humans, such as CYP1A, CYP2D and CYP3A. Objective: To evaluate the hypothesis that the guinea-pig also expresses a functional CYP2C drug-metabolizing enzyme and the P-glycoprotein (P-gp) drug transporter in various tissues. Methods: cDNAs encoding CYP2C and P-gp were obtained from guinea-pig liver or small intestine and sequenced. Western blotting was performed to confirm the expression of CYP2C and P-gp. The functional enzymatic activity of guinea-pig CYP2C was evaluated with microsomal preparations using diclofenac and tolbutamide as specific drug substrates in HPLC analyses. To further study both P-gp and CYP2C functional activities, the guinea-pig ABCB1/MDR1 and CYP2C genes were cloned. The recombinant plasmids were then transfected in HEK293 (human embryonic kidney) cells and either calcein-acetoxymethyl ester (AM) accumulation assays or 14,15-EET/DHET formation experiments were performed to evaluate either P-gp transport activity or CYP2C epoxygenase activity, respectively. The guinea-pig tissue distribution of P-gp was studied by Western blotting. Results: Functional expression of CYP2C was demonstrated in guinea-pig liver microsomal preparations. CYP2C-mediated biotransformation of diclofenac and tolbutamide were shown. Expression of P-gp protein was detected in guinea-pig liver and small intestine. Functional activity of guinea-pig P-gp was demonstrated in ABCB1/MDR1-transfected cells. GP-CYP2C-transfected cells also showed functional epoxygenase activity. Conclusion: The guinea-pig expresses functional CYP2C and P-gp, thus suggesting its usefulness for further validating data obtained with other animal models in drug biotransformation/transport studies. Copyright (c) 2015 John Wiley & Sons, Ltd.
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收藏
页码：183 / 203
页数：21
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