약학회지 (YAKHAK HOEJI)

  • 제58권4호
  • /
  • Pages.255-261
  • /
  • 2014
  • /
  • 0377-9556(pISSN)
  • /
  • 2383-9457(eISSN)
대한약학회 (The Pharmaceutical Society of Korea)
권호 표지

HepaRG 세포를 이용한 Bosentan 약물의 CYP450 효소활성 측정

Measurement of CYP450 Enzymes Activity of Bosentan in HepaRG Cell

  • 한경문 (식품의약품안전평가원 약리연구과) ;
  • 정정아 (식품의약품안전평가원 약리연구과) ;
  • 신지순 (식품의약품안전평가원 약리연구과) ;
  • 차혜진 (식품의약품안전평가원 약리연구과) ;
  • 배영지 (식품의약품안전평가원 약리연구과) ;
  • 김현욱 (식품의약품안전평가원 약리연구과) ;
  • 김영훈 (식품의약품안전평가원 약리연구과) ;
  • 성원근 (식품의약품안전평가원 약리연구과) ;
  • 강호일 (식품의약품안전평가원 약리연구과)
  • Han, Kyoung-Moon (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Jung, Jung-A (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Sin, Ji-Soon (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Cha, Hye-Jin (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Bae, Young-Ji (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Hyun-Uk (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Young-Hoon (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Seong, Won-Keun (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kang, Hoil (Pharmacological Research Division, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 투고 : 2014.07.11
  • 심사 : 2014.08.04
  • 발행 : 2014.08.30
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초록

Poly-pharmacy has been on the rise because of aging of population and chronic disease. Most of drug metabolism happens in the liver by CYP isozymes and the metabolism by CYP450 enzymes. The Cytochrome P450 (CYP) is a superfamily of enzymes that catalyzes the oxidations of many endogenous and exogenous compounds. Primary human Hepatocytes (HH) are considered as the gold standard model for In vitro drug interaction studies. However, there are several limitations (cost, limited life span) for using HH cells. HepaRG cells are being used as a possible alternative. HepaRG cells were cultured in William E medium containing the positive control inducers (1A2: 10, 25, 50 ${\mu}M$ omeprazole, 2C9 and 2C19: 10 ${\mu}M$ rifampin, 3A4: 10, 25, 50 ${\mu}M$ rifampin) at $37^{\circ}C$, 5 % $CO_2$ in a humidified atmosphere. This study was to evaluate the induction of CYP isozymes (1A2, 2C9, 2C19 and 3A4) using LC-MS/MS. We evaluated the potential induction ability of Bosentan, as a drug of pulmonary artery hypertension, in HepaRG cells. For reference, dose of the Bosentan is determined to the basis of the $C_{max}$ (835 mg/ml) value. The enzyme activity demonstrated that CYP2C9 and 3A4 were induced up to 20 times by Bosentan. Like as In vivo, the enzyme activity of CYP2C9 and CYP3A4 is significantly induced in a dose-dependent by Bosentan.

키워드

참고문헌

  1. Brandon, E. F., Raap, C. D., Meijerman, I., Beijnen, J. H. and Schellens, J. H. : An update on in vitro test methods in human hepatic drug biotransformation research: pros and cons. Toxicol. Appl. Pharmacol. 189, 233 (2003). https://doi.org/10.1016/S0041-008X(03)00128-5
  2. McGinnity, D. F., Zhang, G., Kenny, J. R., Hamilton, G. A., Otmani, S., Stams, K. R., Haney, S., Brassil, P., Stresser, D. M. and Riley, R. J. : Evaluation of multiple in vitro systems for assessment of CYP3A4 induction in drug discovery: Human hepatocytes, pregnane X receptor reporter gene, and Fa2N-4 and HepaRG cells. Drug. Metab. Dispos. 37, 1259 (2009). https://doi.org/10.1124/dmd.109.026526
  3. Fahmi, O. A. and Ripp, S. L. : Evaluation of models for prediction drug-drug interactions due to induction. Expert. Opin. Drug. Metab. Toxicol. 6, 1399 (2010). https://doi.org/10.1517/17425255.2010.516251
  4. Drug Interaction Studies - Study design, data analysis, implications for dosing, and labeling recommendations, guidance for industry, FDA, Silver Spring, 2-75 (2012).
  5. Yuan, R., Madani, S., Wei, X. X., Reynolds, K. and Huang, S. M. : Evaluation of cytochrome P450 probe substrates commonly used by the pharmaceutical industry to study in vitro drug interactions. Drug. Metab. Dispos. 30, 1311 (2002). https://doi.org/10.1124/dmd.30.12.1311
  6. Balimane, P. V., Han, Y. H. and Chong, S. : Current industrial practices in assessing CYP450 enzyme induction: Preclinical and clinical. AAPS J. 10, 391 (2008). https://doi.org/10.1208/s12248-008-9037-4
  7. Hewitt, N. J., Lecluyse, E. L. and Ferguson, S. S. : Induction of hepatic cytochrome P450 enzymes: Methods, mechanisms, recommendations, and in vitro-in vivo correlations. Xenobiotica. 37, 1196 (2007). https://doi.org/10.1080/00498250701534893
  8. Rodriguez-Antona, C., Donato, M. T., Boobis, A., Edwards, R. J., Watts, P. S., Castell, J. V. and Gomez-Lechon, M. J. : Cytochrome P450 expression in human hepatocytes and hepatoma cell lines: molecular mechanism that determine lower expression in cultured cells. Xenobiotica. 32, 505 (2002). https://doi.org/10.1080/00498250210128675
  9. Zhang, L., Zhang Y., Zhao, P. and Huang, S. M. : Prediction drug-drug interactions: An FDA perspective. AAPS J. 11, 2 (2009).
  10. Guideline on the investigation of drug interactions, European Medicines Agency, 4-38 (2010).
  11. Antherieu, S., Chesne, C., Li, R., Camus, S., Lahoz, A., Picazo, L., Turpeinen, M., Tolonen, A., Uusitalo, J., Guguen-Guillouzo, C. and Guillouzo, A. : Stable expression, activity and inducibility of cytochromes P450 in differentiated HepaRG cells. Drug. Metab. Dispos. 38, 516 (2010). https://doi.org/10.1124/dmd.109.030197
  12. Guillouzo, A., Corlu, A., Aninat, C., Glaise, D., Morel, F. and Guguen-Guillouzo, C. : The human hepatoma HepaRG cells: A highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. Chem. Biol. Interact. 168, 66 (2007). https://doi.org/10.1016/j.cbi.2006.12.003
  13. Lubberstedt, M., Muller-Vieira, U., Mayer, M., Biemel, K. M., Knospel, F., Knobeloch, D., Nussler, A. K., Gerlach, J. C. and Zeilinger, K. : HepaRG human hepatic cell line utililty as a surrogate for primary human Hepatocytes in drug metabolism assessment in vitro. J. Pharmacol. Toxicol. Methods 63, 59 (2011). https://doi.org/10.1016/j.vascn.2010.04.013
  14. Bowen, W. P., Carey, J. E., Miah, A., McMurray, H. F., Munday, P. W., James, R. S., Coleman, R. A. and Brown, A. M. : Measurement of cytochrome P450 gene induction in human hepatocytes using quantitative real-time reverse transcriptasepolymerase chain reaction. Drug. Metab. Dispos. 28, 781 (2000).
  15. Kanebratt, K. P. and Andersson, T. B. : Evaluation of HepaRG cells as an in vitro model for human drug metabolism studies. Drug. Metab. Dispos. 36, 1444 (2008). https://doi.org/10.1124/dmd.107.020016
  16. Turpeinen, M., Tolonen, A., Chesne, C., Guillouzo, A., Uusitalo, J. and Pelkonen, O. : Functional expression, inhibition and induction of CYP enzymes in HepaRG cells. Toxicol. In Vitro 23, 748 (2009). https://doi.org/10.1016/j.tiv.2009.03.008
  17. Tolonen, A., Petsalo, A., Turpeinen, M., Uusitalo, J. and Pelkonen, O. : In vitro interaction cocktail assay for nine major cytochrome P450 enzymes with 13 probe reactions and a single LC/MSMS run: analytical validation and testing with monoclonal anti-CYP antibodies. J. Mass Spectrom. 42, 960 (2007). https://doi.org/10.1002/jms.1239
  18. Li, X., Chen, X., Li, Q., Wang, L. and Zhong, D. : Validated method for rapid inhibition screening of six cytochrome P450 enzymes by liquid chromatography-tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 852, 128 (2007). https://doi.org/10.1016/j.jchromb.2007.01.006
  19. Wrishko, R. E., Dingemanse, J., Yu, A., Darstein, C., Phillips, D. L. and Mitchell, M. I. : Pharmacokinetic interaction between tadalafil and Bosentan in healthy male subjects. J. Clin. Pharmacol. 48, 610 (2008). https://doi.org/10.1177/0091270008315315
  20. Kenworthy, K. E., Bloomer, J. C., Clarke, S. E. and Houston, J. B. : CYP3A4 drug interactions: correlation of 10 in vitro probes substrates. Br. J. Clin. Pharmacol. 48, 716 (1999).
  21. Fahmi, O. A., Kish, M., Boldt, S. and Obach, R. S. : Cytochrome P450 3A4 mRNA is a more reliable marker than CYP3A4 activity for detecting pregnane X receptor-activated induction of drug-metabolizing enzymes. Drug. Metab. Dispos. 38, 1605 (2010). https://doi.org/10.1124/dmd.110.033126