Abstract
Induction potential of compounds towards CYP1A2, 2B6 and 3A4 via the aryl hydrocarbon (AhR), constitutive androstane (CAR), and pregnane X (PXR) nuclear receptors (NRs), respectively, is routinely determined during small molecule drug development. Significant CYP induction can result in therapeutic failure from clinical exposure of a compound outside the therapeutic window, if the respective enzymes are responsible for a significant portion of the drugs overall metabolism and clearance. Co-medications can also be impacted in a similar manner. Additionally, if metabolism via the induced CYP enzyme results in toxic or pharmacologically active compounds being produced, exaggerated pharmacological effects may be seen resulting in direct and/or indirect toxicity. The following chapter will describe methodologies used for determining CYP induction using isolated human hepatocytes, the current gold standard for such in vitro assays. Where appropriate in the chapter recent guidelines will be highlighted by regulatory agencies, such as, the Food and Drug Administration (FDA) and the European Medicines Agency (EMA).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
US Food and Drug Administration, US Department of Health and Human Services, Center for Drug Evaluation and Research, Center for Biologics Evaluation and Research (CBER) (2012) Guidance for industry: drug interaction studies—study design, data analysis, and implications for dosing and labeling. Silver Spring, MD. http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM292362.pdf
European Medicines Agency (2012) Guideline on the investigation of drug interactions. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2012/07/WC500129606.pdf
Amacher DE (2010) The effects of cytochrome P450 induction by xenobiotics on endobiotic metabolism in pre-clinical safety Studies. Toxicol Mech Methods 20(4):159–166
Gomez-Lechon MJ, Donato T, Jover R et al (2001) Expression and induction of a large set of drug-metabolizing enzymes by the highly differentiated human hepatoma cell line BC2. Eur J Biochem 268(5):1448–1459
Westerink WM, Schoonen WG (2007) Cytochrome P450 enzyme levels in HepG2 cells and cryopreserved primary human hepatocytes and their induction in HepG2 cells. Toxicol In Vitro 21(8):1581–1591
Hariparsad N, Carr BA, Evers R, Chu X (2008) Comparison of immortalized Fa2N-4 cells and human hepatocytes as in vitro models for cytochrome P450 induction. Drug Metab Dispos 36(6):1046–1055
McGinnity DF, Zhang G, Kenny JR et al (2009) 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(6):1259–1268
Gerets HH, Tilmant K, Gerin B et al (2012) Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol Toxicol 28(2):69–87
Fahmi OA, Ripp SL (2010) Evaluation of models for predicting drug-drug interactions due to induction. Models for predicting drug-drug interactions due to induction. Expert Opin Drug Metab Toxicol 6(11):1399–1416
Chu V, Einolf HJ, Evers R et al (2009) In vitro and in vivo induction of cytochrome p450: a survey of the current practices and recommendations: a pharmaceutical research and manufacturers of America perspective. Drug Metab Dispos 37(7):1339–1354
Hewitt NJ, Lecluyse EL, Ferguson SS (2007) Induction of hepatic cytochrome P450 enzymes: methods, mechanisms, recommendations, and in vitro-in vivo correlations. Xenobiotica 37(10–11):1196–1224
Parkinson A, Mudra DR, Johnson C, Dwyer A, Carroll KM (2004) The effects of gender, age, ethnicity, and liver cirrhosis on cytochrome P450 enzyme activity in human liver microsomes and inducibility in cultured human hepatocytes. Toxicol Appl Pharmacol 199(3):193–209
Roymans D, Annaert P, Van Houdt J et al (2005) Expression and induction potential of cytochromes P450 in human cryopreserved hepatocytes. Drug Metab Dispos 33(7):1004–1016
Smith CM, Nolan CK, Edwards MA, Hatfield JB, Stewart TW, Ferguson SS, LeCluyse EL, Sahi J (2012) A comprehensive evaluation of metabolic activity and intrinsic clearance in suspensions and monolayer cultures of cryopreserved primary human hepatocytes. J Pharm Sci 101(10):3989–4002
Varma MV, Lin J, Rotter CJ, Fahmi OA, Lam J, El-Kattan AF, Goosen TC, Lai Y (2013) Quantitative prediction of repaglinide-rifampicin complex drug interactions using dynamic and static mechanistic models: delineating differential CYP3A4 induction and OATP1B1 inhibition potential of rifampicin. Drug Metab Dispos 41(5):966–974
Acknowledgements
The views expressed here are solely those of the authors and do not reflect the opinions of Janssen Research & Development, LLC.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Singer, M., Sensenhauser, C., Dallas, S. (2014). In Vitro CYP Induction Using Human Hepatocytes. In: Caldwell, G., Yan, Z. (eds) Optimization in Drug Discovery. Methods in Pharmacology and Toxicology. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-742-6_14
Download citation
DOI: https://doi.org/10.1007/978-1-62703-742-6_14
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-741-9
Online ISBN: 978-1-62703-742-6
eBook Packages: Springer Protocols