title: Application of lipid peroxidation products as biomarkers for flutamide-induced oxidative stress in vitro.
abstract: The early identification of hepatotoxicity is a fundamental goal of preclinical safety studies in drug discovery and early development. Sensitive biomarkers warrant the determination of potential underlying mechanisms that help characterizing a disruption of physiological conditions prior to cell death. This study shows the potential of different lipid peroxidation products, namely isoprostanes and hydroxynonenal (HNE) derivatives, to serve as early safety biomarkers of hepatotoxicity caused by oxidative stress as underlying mechanism. The hepatotoxic drug flutamide was used as model compound in primary hepatocytes. Incubation conditions were optimized by the addition of hydrogen peroxide generating substrates enhancing the cellular response upon oxidative stress. A time and dose dependent response of different isoprostanes and prostaglandins (15R-prostaglandin D2, prostaglandin E2, 13,14-dihydro-15-keto prostaglandin E2 and 5‑iso prostaglandin F2α-VI) became manifest after 6 and 24h of treatment in 3.8- to 17.4-fold increased concentrations where no overt hepatocellular damage was observed. For HNE-mercapturic acid and its metabolite dihydroxynonene-mercapturic acid a similar response was evident with a 20- and 10-fold increase from control after 24 h of treatment, respectively. These data indicate that lipid peroxidation products as markers of reactive oxygen species are more sensitive than conventional cytotoxicity markers for an early detection of drug-induced liver injury.
authors.affiliation: Pharmaceutical Sciences, Pharma Research and Early Development, pRED, Roche Innovation Center Basel, 4070 Basel, Switzerland; Institute of Molecular Pharmacy, University of Basel, Klingenbergstrasse 50, CH-4040 Basel, Switzerland. Electronic address: marieke.teppner@evotec.com.
authors.forename: Marieke
authors.initials: M
authors.lastname: Teppner
authors.affiliation: Pharmaceutical Sciences, Pharma Research and Early Development, pRED, Roche Innovation Center Basel, 4070 Basel, Switzerland.
authors.forename: Franziska
authors.initials: F
authors.lastname: Böss
authors.affiliation: Institute of Molecular Pharmacy, University of Basel, Klingenbergstrasse 50, CH-4040 Basel, Switzerland.
authors.forename: Beat
authors.initials: B
authors.lastname: Ernst
authors.affiliation: Pharmaceutical Sciences, Pharma Research and Early Development, pRED, Roche Innovation Center Basel, 4070 Basel, Switzerland. Electronic address: axel.paehler@roche.com.
authors.forename: Axel
authors.initials: A
authors.lastname: Pähler
mesh_terms: D000726:Androgen Antagonists / Q000009:adverse effects*; D000818:Animals; D015415:Biomarkers; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005485:Flutamide / Q000009:adverse effects*; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D028421:Isoprostanes / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects*; D018384:Oxidative Stress; D051381:Rats; D013997:Time Factors
chemical_list: D000726:Androgen Antagonists; D015415:Biomarkers; D028421:Isoprostanes; D005485:Flutamide; D006861:Hydrogen Peroxide
keywords: Hepatotoxicity; Hydroxynonenal; Isoprostanes; Lipid peroxidation; Oxidative stress; Primary hepatocytes