title: Promising toxicological biomarkers for the diagnosis of liver injury types: Bile acid metabolic profiles and oxidative stress marker as screening tools in drug development.
abstract: Promising biomarkers were identified in adult male Crl:CD (SD) rats for the screening of new chemical entities for their potential to cause liver injury. We examined the serum biochemistry, liver histopathology, and bile acid profiles by LC-MS/MS, and the mRNA expression of transporters and CYPs by an RT-PCR after the following treatments to male Crl:CD (SD) rats: (a) bile duct ligation (BDL); (b) a single oral dose of 150 mg/kg α-naphthylisothiocyanate (ANIT); and (c) repeated oral doses of a novel pyrrolidinecarboxylic acid derivative (abbreviated as PCA) at 30, 300, and 1000 mg/kg. The serum total bile acid levels and bilirubin concentrations were found to be elevated in all of the groups. However, the bile acid component profiles of the PCA group differed significantly from BDL and ANIT models: deoxycholic acid, lithocholic acid, and sulfated bile acids were upregulated in a dose-dependent manner only in the PCA group. In addition, the PCA group demonstrated high levels of hepatic heme oxygenase-1 expression, whereas the profiles of the mRNA levels of the hepatic transporters and CYPs of all groups were found to be similar. The histopathological findings, for both the BDL and ANIT groups, were of bile duct hyperplasia, hepatocyte degeneration and necrosis. In contrast, only bile duct hyperplasia and hepatocyte degeneration were observed in the PCA group, even at a lethal dose. These results indicated that PCA induced a cholestatic condition and the increase of oxidative stress markers implies that this will also lead hepatocellular injury. In conclusion, the serum bile acid components and sulfated bile acid levels, and the expression of oxidative stress markers could provide information that aids in the diagnosis of liver injury type and helps to elucidate the mechanisms of hepatotoxicity. These findings can be extrapolated into our clinical investigation. The analysis of these crucial biomarkers is likely to be a useful screening tool in the lead optimization phase of drug discovery.
authors.affiliation: Drug Metabolism & Pharmacokinetics Research Laboratories, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, 140-8710, Japan. Electronic address: masubuchi.noriko.wa@daiichisankyo.co.jp.
authors.forename: Noriko
authors.initials: N
authors.lastname: Masubuchi
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd., 1-16-13, Kita-Kasai, Edogawa-ku, 134-8630, Tokyo, Japan.
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Nishiya
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd., 1-16-13, Kita-Kasai, Edogawa-ku, 134-8630, Tokyo, Japan.
authors.forename: Masako
authors.initials: M
authors.lastname: Imaoka
authors.affiliation: Daiichi Sankyo RD Novare, 1-16-13, Kita-Kasai, Edogawa-ku, 134-8630, Tokyo, Japan.
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Mizumaki
authors.affiliation: Drug Metabolism & Pharmacokinetics Research Laboratories, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, 140-8710, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Okazaki
mesh_terms: D015058:1-Naphthylisothiocyanate / Q000008:administration & dosage / Q000633:toxicity*; D000818:Animals; D001647:Bile Acids and Salts / Q000097:blood / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis / Q000097:blood / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000097:blood / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D002779:Cholestasis / Q000097:blood / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D055442:Metabolome / Q000187:drug effects*; D018384:Oxidative Stress / Q000187:drug effects*; D011761:Pyrrolidonecarboxylic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D001647:Bile Acids and Salts; D015415:Biomarkers; D015058:1-Naphthylisothiocyanate; D003577:Cytochrome P-450 Enzyme System; D011761:Pyrrolidonecarboxylic Acid
keywords: CYP; Cholestasis; Deoxycholic acid; Hepatic transporter; Ho-1; Lithocholic acid