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Evaluation of Drug Metabolism, Drug–Drug Interactions, and In Vitro Hepatotoxicity with Cryopreserved Human Hepatocytes

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Hepatocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 640))

Abstract

Human-based in vitro hepatic experimental systems are now used routinely in drug development. The initial concept of the use of human-based in vitro systems is based on the known species–species differences in drug properties. Human-specific drug properties, by definition, cannot be defined using nonhuman experimental animals and therefore can be only assessed in the preclinical phase of drug development using in vitro human-based experimental systems such as human hepatocytes. Successful cryopreservation of human hepatocytes greatly enhances the utility of this valuable in vitro experimental system, allowing storage, transport, convenient scheduling of experimentation, and repeat experimentation using hepatocytes isolated from the same donors. Assay procedures with cryopreserved human hepatocytes using multiwell plates for the evaluation of critical drug properties including metabolic stability, drug–drug interaction potential, and drug toxicity during drug development are described.

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References

  1. DiMasi, J.A., Hansen, R.W., and Grabowski, H.G. (2003) The price of innovation: new estimates of drug development costs. J. Health Econ. 22, 151–185.

    Article  PubMed  Google Scholar 

  2. Guengerich, F.P. (2006) Cytochrome P450 s and other enzymes in drug metabolism and toxicity. AAPS J. 8, E101–E111.

    Article  PubMed  CAS  Google Scholar 

  3. Kola, I. and Landis, J. (2004) Can the pharmaceutical industry reduce attrition rates? Nat. Rev. Drug Discov. 3, 711–715.

    Article  PubMed  CAS  Google Scholar 

  4. Fry, J.R. (1982) The metabolism of drugs by isolated hepatocytes. Q. Rev. Drug Metab. Drug Interact. 4, 99–122.

    Article  PubMed  CAS  Google Scholar 

  5. Li, A.P. (2007) Human hepatocytes: isolation, cryopreservation and applications in drug development. Chem. Biol. Interact. 168, 16–29.

    Article  PubMed  CAS  Google Scholar 

  6. Gomez-Lechon, M.J., Castell, J.V., and Donato, M.T. (2007) Hepatocytes – the choice to investigate drug metabolism and toxicity in man: in vitro variability as a reflection of in vivo. Chem. Biol. Interact. 168, 30–50.

    Article  PubMed  CAS  Google Scholar 

  7. Li, A.P. (2004) In vitro approaches to evaluate ADMET drug properties. Curr. Top. Med. Chem. 4, 701–706.

    Article  PubMed  CAS  Google Scholar 

  8. Li, A.P. (2001) Screening for human ADME/Tox drug properties in drug discovery. Drug Discov. Today 6, 357–366.

    Article  PubMed  CAS  Google Scholar 

  9. Hewitt, N.J., Lechon, M.J., Houston, J.B., Hallifax, D., Brown, H.S., Maurel, P., Kenna, J.G., Gustavsson, L., Lohmann, C., Skonberg, C., Guillouzo, A., Tuschl, G., Li, A.P., LeCluyse, E., Groothuis, G.M., and Hengstler, J.G. (2007) Primary hepatocytes: current understanding of the regulation of metabolic enzymes and transporter proteins, and pharmaceutical practice for the use of hepatocytes in metabolism, enzyme induction, transporter, clearance, and hepatotoxicity studies. Drug Metab. Rev. 39, 159–234.

    Article  PubMed  CAS  Google Scholar 

  10. Loretz, L.J., Li, A.P., Flye, M.W., and Wilson, A.G. (1989) Optimization of cryopreservation procedures for rat and human hepatocytes. Xenobiotica 19, 489–498.

    Article  PubMed  CAS  Google Scholar 

  11. Li, A.P., Lu, C., Brent, J.A., Pham, C., Fackett, A., Ruegg, C.E., and Silber, P.M. (1999b) Cryopreserved human hepatocytes: characterization of drug-metabolizing enzyme activities and applications in higher throughput screening assays for hepatotoxicity, metabolic stability, and drug–drug interaction potential. Chem. Biol. Interact. 121, 17–35.

    Article  PubMed  CAS  Google Scholar 

  12. Li, A.P., Gorycki, P.D., Hengstler, J.G., Kedderis, G.L., Koebe, H.G., Rahmani, R., de Sousas, G., Silva, J.M., and Skett, P. (1999a) Present status of the application of cryopreserved hepatocytes in the evaluation of xenobiotics: consensus of an international expert panel. Chem. Biol. Interact. 121, 117–123.

    Article  PubMed  CAS  Google Scholar 

  13. Jouin, D., Blanchard, N., Alexandre, E., Delobel, F., David-Pierson, P., Lavé, T., Jaeck, D., Richert, L., and Coassolo, P. (2006) Cryopreserved human hepatocytes in suspension are a convenient high throughput tool for the prediction of metabolic clearance. Eur. J. Pharm. Biopharm. 63, 347–355.

    Article  PubMed  CAS  Google Scholar 

  14. Brown, H.S., Griffin, M., and Houston, J.B. (2007) Evaluation of cryopreserved human hepatocytes as an alternative in vitro system to microsomes for the prediction of metabolic clearance. Drug Metab. Dispos. 35, 293–301.

    Article  PubMed  CAS  Google Scholar 

  15. Kafert-Kasting, S., Alexandrova, K., Barthold, M., Laube, B., Friedrich, G., Arseniev, L., and Hengstler, J.G. (2006) Enzyme induction in cryopreserved human hepatocyte cultures. Toxicology 220, 117–125.

    Article  PubMed  CAS  Google Scholar 

  16. Bi, Y.A., Kazolias, D., and Duignan, D.B. (2006) Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport. Drug Metab. Dispos. 34, 1658–1665.

    Article  PubMed  CAS  Google Scholar 

  17. Lavé, T., Dupin, S., Schmitt, C., Valles, B., Ubeaud, G., Chou, R.C., Jaeck, D., and Coassolo, P. (1997) The use of human hepatocytes to select compounds based on their expected hepatic extraction ratios in humans. Pharm. Res. 14, 152–155.

    Article  PubMed  Google Scholar 

  18. Lu, C., Li, P., Gallegos, R., Uttamsingh, V., Xia, C.Q., Miwa, G.T., Balani, S.K., and Gan, L.S. (2006) Comparison of intrinsic clearance in liver microsomes and hepatocytes from rats and humans: evaluation of free fraction and uptake in hepatocytes. Drug Metab. Dispos. 34, 1600–1605.

    Article  PubMed  CAS  Google Scholar 

  19. Lee, K., Vandenberghe, Y., Herin, M., Cavalier, R., Beck, D., Li, A., Verbeke, N., Lesne, M., and Roba, J. (1994) Comparative metabolism of SC-42867 and SC-51089, two PGE2 antagonists, in rat and human hepatocyte cultures. Xenobiotica 24, 25–36.

    Article  PubMed  CAS  Google Scholar 

  20. Zhang, D., Wang, L., Raghavan, N., Zhang, H., Li, W., Cheng, P.T., Yao, M., Zhang, L., Zhu, M., Bonacorsi, S., Yeola, S., Mitroka, J., Hariharan, N., Hosagrahara, V., Chandrasena, G., Shyu, W.C., and Humphreys, W.G. (2007) Comparative metabolism of radiolabeled muraglitazar in animals and humans by quantitative and qualitative metabolite profiling. Drug Metab. Dispos. 35, 150–167.

    Article  PubMed  CAS  Google Scholar 

  21. FDA. (2006) Drug interaction studies―study design, data analysis, and implications for dosing and labeling. Available at: http://www.fda.gov/cder/guidance/2635fnl.pdf.

  22. Li, A.P. (2009) Metabolism comparative cytotoxicity assay (MCCA) and cytotoxic metabolic pathway identification assay (CMPIA) with cryopreserved human hepatocytes for the evaluation of metabolism-based cytotoxicity in vitro: proof-of-concept study with aflatoxin B1. Chem. Biol. Interact. 179, 4–8.

    Article  PubMed  CAS  Google Scholar 

  23. Li, A.P., Bode, C., and Sakai, Y. (2004) A novel in vitro system, the integrated discrete multiple organ cell culture (IdMOC) system, for the evaluation of human drug toxicity: comparative cytotoxicity of tamoxifen towards normal human cells from five major organs and MCR-7 adenocarcinoma breast cancer cells. Chem. Biol. Interact. 150, 129–136.

    Article  PubMed  CAS  Google Scholar 

  24. Li, A.P. (2008) In vitro evaluation of human xenobiotic toxicity: scientific concepts and the novel integrated discrete multiple cell co-culture (IdMOC) technology. ALTEX 25, 4F–49.

    PubMed  CAS  Google Scholar 

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Correspondence to Albert P. Li .

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Li, A.P. (2010). Evaluation of Drug Metabolism, Drug–Drug Interactions, and In Vitro Hepatotoxicity with Cryopreserved Human Hepatocytes. In: Maurel, P. (eds) Hepatocytes. Methods in Molecular Biology, vol 640. Humana Press. https://doi.org/10.1007/978-1-60761-688-7_15

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  • DOI: https://doi.org/10.1007/978-1-60761-688-7_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-687-0

  • Online ISBN: 978-1-60761-688-7

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