Summary
Uridine 5′-diphospho-N-acetylgalactosamine glycosyltransferases (UGTs) and sulfotransferases (SULTs) are 2 phase II enzymes that are actively involved in detoxification processes as well as in drug metabolism. Compared with cytochrome P450 enzymes, the role of UGTs and SULTs in drug metabolism has received little attention. Liver microsomes, S9 fractions, and cryopreserved hepatocytes from human, dog, cynomolgus monkey, mouse, and rat were used as matrices in the study. Single compound, 7-hydroxycoumarin (7-HC), along with necessary cofactors was dosed into the matrices and incubated at 37° C; formation of two metabolites, 7-HC-glucuronide and 7-HC-sulfate, was determined with liquid chromatography with tandem mass spectrometry. Within the same species, the UGTs activities in microsomes and S9 fractions were comparable. In addition, UGTs activities in cryopreserved hepatocytes were lower than in the other matrices. Also, the SULTs activities were much higher in S9 fractions than in cryopreserved hepatocytes and microsomes. Species differences on UGTs and SULTs activities were also observed. The results indicated that S9 fractions, microsomes, and cryopreserved hepatocytes might be useful for UGTs metabolism study, whereas S9 fractions appear to be the most appropriate matrix for both UGTs and SULTs metabolism. Species differences with respect to phase II metabolism also need to be taken into consideration when selecting an in vitro system to evaluate various aspects of drug metabolism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abbott, F. V.; Palmour, R. M. Morphine-6-glucuronide: analgesic effects and receptor binding profile in rats. Life Sci. 43(21):1685–1695; 1988.
Bock, K. W. Vertebrate UDP-glucuronosyltransferases: functional and evolutionary aspects. Biochem. Pharmacol. 66(5):691–696; 2003.
Donato, M. T.; Castell, J. V. Strategies and molecular probes to investigate the role of cytochrome P450 in drug metabolism: focus on in vitro studies. Clin. Pharmacokinet. 42(2):153–178; 2003.
Ekins, S.; Murray, G. I.; Burke, M. D.; Williams, J. A.; Marchant, N. C.; Hawksworth, G. M. Quantitative differences in phase I and II metabolism between rat precision-cut liver slices and isolated hepatocytes. Drug Metab. Dispos. 23(11):1274–1279; 1995.
Ekins, S.; Williams, J. A.; Murray, G. I.; Burke, M. D.; Marchant, N. C.; Engeset, J.; Hawksworth, G. M. Xenobiotic metabolism in rat, dog, and human precision-cut liver slices, freshly isolated hepatocytes, and vitrified precision-cut liver slices. Drug Metab. Dispos. 24(9):990–995; 1996.
Falany, C. N.; Kerl, E. A. Sulfation of minoxidil by human liver phenol sulfotransferase. Biochem. Pharmacol. 40(5):1027–1032; 1990.
Farrell, D. F.; McKhann, G. M. Characterization of cerebroside sulfotransferase from rat brain. J. Biol. Chem. 246(15):4694–4702; 1971.
Garcia, M.; Rager, J.; Wang, Q.; Strab, R.; Hidalgo, I. J.; Owen, A.; Li, J. Cryopreserved human hepatocytes as alternative in vitro model for cytochrome p450 induction studies. In Vitro Cell. Dev. Biol. 39A(7):283–287; 2003.
Gervasini, G.; Carrillo, J. A.; Benitez, J. Potential role of cerebral cytochrome p450 in clinical pharmacokinetics: modulation by endogenous compounds. Clin. Pharmacokinet. 43(11):693–706; 2004.
Goldstein, J. A.; Faletto, M. B. Advances in mechanisms of activation and deactivation of environmental chemicals. Environ. Health Perspect. 100:169–176; 1993.
Hauser, S. C.; Ziurys, J. C.; Gollan, J. L. A membrane transporter mediates access of uridine 5′-diphosphoglucuronic acid from the cytosol into the endoplasmic reticulum of rat hepatocytes: implications for glucuronidation reactions. Biochim. Biophys. Acta 967(2):149–157; 1988.
Hengstler, J. G.; Utesch, D.; Steinberg, P., et al. Cryopreserved primary hepatocytes as a constantly available in vitro model for the evaluation of human and animal drug metabolism and enzyme induction. Drug Metab. Rev. 32(1):81–118; 2000.
Iyer, K. R.; Sinz, M. W. Characterization of phase I and phase II hepatic drug metabolism activities in a panel of human liver preparations. Chem. Biol. Interact. 118(2):151–169; 1999.
Klaassen, C. D.; Liu, L.; Dunn, R. T., II. Regulation of sulfotransferase mRNA expression in male and female rats of various ages. Chem. Biol. Interact. 109(1–3):299–313; 1998.
Mackenzie, P. I.; Owens, I. S.; Burchell, B., et al. The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. Pharmacogenetics 7(4):255–269; 1997.
Muraoka, M.; Ishida, N. Molecular characterization of human UDP-glucuronic acid/UDP-N-acetylgalactosamine transporter, a novel nucleotide sugar transporter with dual substrate specificity. FEBS Lett. 495(1–2):87–93; 2001.
Otake, Y.; Hsieh, F.; Walle, T. Glucuronidation versus oxidation of the flavonoid galangin by human liver microsomes and hepatocytes. Drug Metab. Dispos. 30(5):576–581; 2002.
Ouyang, Y.; Lane, W. S.; Moore, K. L. Tyrosylprotein sulfotransferase: purification and molecular cloning of an enzyme that catalyzes tyrosine O-sulfation, a common posttranslational modification of eukaryotic proteins. Proc. Natl. Acad. Sci. USA 95(6):2896–2901; 1998.
Ouyang, Y. B.; Moore, K. L. Molecular cloning and expression of human and mouse tyrosylprotein sulfotransferase-2 and a tyrosylprotein sulfotransferase homologue in Caenorhabditis elegans. J. Biol. Chem. 273(38):24770–24774; 1998.
Rauchschwalbe, S. K.; Zuhlsdorf, M. T.; Wensing, G.; Kuhlmann, J. Glucuronidation of acetaminophen is independent of UGT1A1 promotor genotype. Int. J. Clin. Pharmacol. Ther. 42(2):73–77; 2004.
Roymans, D.; Van Looveren, C.; Leone, A., et al. Determination of cytochrome P540 1A2 and cytochrome P450 3A4 induction in cryopreserved human hepatocytes. Biochem. Pharmacol. 67(3):427–437; 2004.
Sherer, J. E.; Shipley, L. A.; Vandenbranden, M. R.; Binkley, S. N.; Wrighton, S. A. Comparisons of phase I and phase II in vitro hepatic enzyme activities of human, dog, rhesus monkey, and cynomolgus monkey. Drug Metab. Dispos. 23(11):1231–1241; 1995.
Snodgrass, B.; Gagne, P.; Picano, P., et al. Comparison of major phase I and phase II metabolism reactions in cryopreserved, dog and human hepatocytes. Drug Metab. Rev. 35(S2): Abstract 73, 2003. URL: http://www.dekker.com/servlet/product/DOI/101081DMR120026877.
Swales, N. J.; Utesch, D. Metabolic activity of fresh and cryopreserved, dog hepatocyte suspensions. Xenobiotica 28:937–948; 1998.
Tephly, T. R.; Burchell, B. UDP-glucuronyltransferases: a family of detoxifying enzymes. Trends Pharmacol. Sci. 11:276–279; 1990.
Thohan, S.; Zurich, M. C.; Chung, H.; Weiner, M.; Kane, A. S.; Rosen, G. M. Tissue slices revisited: evaluation and development of a short-term incubation for integrated drug metabolism. Drug Metab. Dispos. 29(10):1337–1342; 2001.
Vaidyanathan, J. B.; Walle, T. Glucuronidation and sulfation of the tea flavonoid (−)-epicatechnin by the human and rat enzymes. Drug Metab. Dispos. 30(8):897–903; 2002.
Weinshilboum, R. M.; Otterness, D. M.; Aksoy, I. A.; Wood, T. C.; Her, C.; Raftogianis, R. B. Sulfation and sulfotransferases 1: sulfotransferase molecular biology: cDNAs and genes. FASEB J. 11(1):3–14; 1997.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, Q., Jia, R., Ye, C. et al. Glucuronidation and sulfation of 7-hydroxycoumarin in liver matrices from human, dog, monkey, rat, and mouse. In Vitro Cell.Dev.Biol.-Animal 41, 97–103 (2005). https://doi.org/10.1290/0501005.1
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1290/0501005.1