title: De-risking bio-therapeutics for possible drug interactions using cryopreserved human hepatocytes.
abstract: Inflammatory diseases such as rheumatoid arthritis and psoriasis are characterized by increases in circulating cytokines, which play an important role in modulation of the disease state. Several marketed bio-therapeutics target cytokines and act as effective treatment strategies. Previous in-vitro and in-vivo studies have suggested that cytokines may have both direct and indirect effects on drug metabolizing enzyme levels in the liver. Few studies have characterized models to evaluate the risk of potential drug interactions that might be mediated by changes in cytokine levels. In the present studies the potential of three cytokines (IL-2, IL-6 and TNF-α) to modulate gene expression and activity of the major human cytochrome P450 (CYP) enzymes (CYP1A2, 2B6, 2C9, 2C19, 2D6, and 3A4) in cryopreserved human hepatocytes (CHH) was investigated. Significant decreases in the activity of all 6 CYP isoforms occurred in hepatocytes incubated with TNF-α or IL-6 (17-85%; and 22-76% of untreated control values, respectively). TNF-α down-regulated the gene expression of CYP1A2, 2D6 and 3A4 only, whereas IL-6 down-regulated gene expression of all of the tested CYP isoforms except 2D6. IL-2 had only mild effects on CYP activity and mRNA levels of examined isoforms. In CHH exposed to TNF-α, changes in CYP activity were not always paralleled by gene expression alterations for three of the examined CYP isoforms. These studies highlight several potential pitfalls in using isolated human hepatocytes for determination of drug interactions by bio-therapeutics including lack of correlation of mRNA and activity measurements for some CYP isoforms when using single time point determinations, and appropriateness of the model for indirect acting cytokine and cytokine modulators.
authors.affiliation: Janssen Research and Development, Drug Safety Sciences, 1000 Route 202 South, Raritan, New Jersey 08869, USA. sdallas@its.jnj.com
authors.forename: Shannon
authors.initials: S
authors.lastname: Dallas
authors.forename: Carlo
authors.initials: C
authors.lastname: Sensenhauser
authors.forename: Ameesha
authors.initials: A
authors.lastname: Batheja
authors.forename: Monica
authors.initials: M
authors.lastname: Singer
authors.forename: Maria
authors.initials: M
authors.lastname: Markowska
authors.forename: Cindy
authors.initials: C
authors.lastname: Zakszewski
authors.forename: Rao N V S
authors.initials: RN
authors.lastname: Mamidi
authors.forename: Michael
authors.initials: M
authors.lastname: McMillia
authors.forename: Chao
authors.initials: C
authors.lastname: Han
authors.forename: Honghui
authors.initials: H
authors.lastname: Zhou
authors.forename: Jose
authors.initials: J
authors.lastname: Silva
mesh_terms: D001688:Biological Products / Q000378:metabolism* / Q000494:pharmacology; D002675:Child, Preschool; D015925:Cryopreservation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004347:Drug Interactions / Q000502:physiology*; D005260:Female; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012307:Risk Factors
chemical_list: D001688:Biological Products; D016207:Cytokines; D003577:Cytochrome P-450 Enzyme System