Enzymatic oxidation of phthalazine with guinea pig liver aldehyde oxidase and liver slices: inhibition by isovanillin

• Authors: Georgios I Panoutsopoulos , Christine Beedham
• Languages of publication: EN

Abstracts

EN

The enzymes aldehyde oxidase and xanthine oxidase catalyze the oxidation of a wide range of N-heterocycles and aldehydes. These enzymes are widely known for their role in the metabolism of N-heterocyclic xenobiotics where they provide a protective barrier by aiding in the detoxification of ingested nitrogen-containing heterocycles. Isovanillin has been shown to inhibit the metabolism of aromatic aldehydes by aldehyde oxidase, but its inhibition towards the heterocyclic compounds has not been studied. The present investigation examines the oxidation of phthalazine in the absence and in the presence of the inhibitor isovanillin by partially purified aldehyde oxidase from guinea pig liver. In addition, the interaction of phthalazine with freshly prepared guinea pig liver slices, both in the absence and presence of specific inhibitors of several liver oxidizing enzymes, was investigated. Aldehyde oxidase rapidly converted phthalazine into 1-phthalazinone, which was completely inhibited in the presence of isovanillin (a specific inhibitor of aldehyde oxidase). In freshly prepared liver slices, phthalazine was also rapidly converted to 1-phthalazinone. The formation of 1-phthalazinone was completely inhibited by isovanillin, whereas disulfiram (a specific inhibitor of aldehyde dehydrogenase) only inhibited 1-phthalazinone formation by 24% and allopurinol (a specific inhibitor of xanthine oxidase) had little effect. Therefore, isovanillin has been proved as an inhibitor of the metabolism of heterocyclic substrates, such as phthalazine, by guinea pig liver aldehyde oxidase, since it had not been tested before. Thus it would appear from the inhibitor results that aldehyde oxidase is the predominant enzyme in the oxidation of phthalazine to 1-phthalazinone in freshly prepared guinea pig liver slices, whereas xanthine oxidase only contributes to a small extent and aldehyde dehydrogenase does not take any part.

Keywords

EN

xanthine oxidase   phthalazine   aldehyde oxidase   liver slices   disulfiram   isovanillin   allopurinol  

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2004
• Volume: 51
• Issue: 4
• Pages: 943-951

Dates

published

2004

received

2004-04-05

revised

2004-06-24

accepted

2004-07-07

References

• Beedham C. (1985) Molybdenum hydroxylases as drug-metabolizing enzymes. Drug Metab Rev.; 16: 119-56.
• Beedham C. (1987) Molybdenum hydroxylases: biological distribution and substrate-inhibitor specificity. In Progress in Medicinal Chemistry. Ellis GP, West GB. eds, vol. 24, pp. 85-127, Elsevier, Amsterdam.
• Beedham C, Bruce SE, Critchley DJ, Al-Tayib Y, Rance DJ. (1987) Species variation in hepatic aldehyde oxidase activity. Eur J Drug Metab Pharmacokinet.; 12: 307-10.
• Beedham C, Bruce SE, Critchley DJ, Rance DJ. (1990) 1-Substituted phthalazines as probes of the substrate-binding site of mammalian molybdenum hydroxylases. Biochem Pharmacol.; 39: 1213-21.
• Beedham C, Critchley DJP, Rance DJ. (1995a) Substrate specificity of human liver aldehyde oxidase toward substituted quinazolines and phthalazines: a comparison with hepatic enzyme from guinea pig, rabbit and baboon. Arch Biochem Biophys.; 319: 481-90.
• Beedham C, Peet CF, Panoutsopoulos GI, Carter H, Smith JA. (1995b) Role of aldehyde oxidase in biogenic amine metabolism. Prog Brain Res.; 106: 345-53.
• Bradford MM. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem.; 72: 248-54.
• Bruder G, Heid H, Jarasch ED, Keenan TW, Mather IH. (1982) Characteristics of membrane-bound and soluble forms of xanthine oxidase from milk and endothelial cells of capillaries. Biochim Biophys Acta.; 701: 357-69.
• Critchley DJ, Rance DJ, Beedham C. (1992) Subcellular localization of guinea pig hepatic molybdenum hydroxylases. Biochem Biophys Res Commun.; 185: 54-9.
• Deitrich RA, Erwin VG. (1971) Mechanism of the inhibition of aldehyde dehydrogenase in vivo by disulfiram and diethyldithiocarbamate. Mol Pharmacol.; 7: 301-7.
• De Kanter R, Olinga P, De Jager MH, Merema MT, Meijer DKF, Groothius GMM. (1999) Organ slices as an in vitro test system for drug metabolism in human liver, lung and kidney. Toxicol in Vitro.; 13: 737-44.
• Dennis N, Katritsky AR, Ramaiah M. (1975) 1,3-Dipolar character of six-membered aromatic rings. Part X. Pyridazine and benzopyridazine betaines. J Chem Soc Perkin Trans.; 1: 1506-14.
• Ekins S. (1996a) Short-term maintainance of phase I and II metabolism in precision-cut liver slices in dynamic organ culture. Drug Metab Dispos.; 24: 364-6.
• Ekins S. (1996b) Past, present and future applications of precision-cut liver slices for in vitro xenobiotic metabolism. Drug Metab Rev.; 28: 591-623.
• Hille R, Massey V. (1981) Tight binding inhibitors of xanthine oxidase. Pharmacol Ther.; 14: 249-63.
• Johnson C, Stubley-Beedham C, Stell JGP. (1984) Elevation of molybdenum hydroxylase levels in rabbit liver after ingestion of phthalazine or its hydroxylated metabolite. Biochem Pharmacol.; 33: 3699-705.
• Krenitsky TA. (1978) Aldehyde oxidase and xanthine oxidase: functional and evolutionary relationships. Biochem Pharmacol.; 27: 2763-4.
• Krenitsky TA, Neil SM, Elion GB, Hitchings GH. (1972) A comparison of the specificities of xanthine oxidase and aldehyde oxidase. Arch Biochem Biophys.; 150: 585-99.
• Krumdieck CL, Dos Santos JE, Ho KJ. (1980) A new instrument for the rapid preparation of tissue slices. Anal Biochem.; 104: 118-23.
• Lerche-Langrand C, Toutain HJ. (2000) Precision-cut liver slices: characteristics and use for in vitro pharmaco-toxicology. Toxicology.; 153: 221-53.
• Lipsky JJ, Shen ML, Naylor S. (2001) Overview - In vitro inhibition of aldehyde dehydrogenase by disulfiram and metabolites. Chem-Biol Interact.; 130-132: 81-91.
• Moriwaki Y, Yamamoto T, Nasako Y, Takahashi S, Suda M, Hiroishi K, Hada T, Higashino K. (1993) In vitro oxidation of pyrazinamide and allopurinol by rat liver aldehyde oxidase. Biochem Pharmacol.; 46: 975-81.
• Panoutsopoulos GI, Beedham C. (2005) Enzymatic oxidation of vanillin, isovanillin and protocatechnic aldehyde with freshly prepared guinea pig liver slices. Cell Physiol Biochem.; 15: in press..
• Panoutsopoulos GI, Beedham C. (2004) Kinetics and specificity of guinea pig liver aldehyde oxidase and bovine milk xanthine oxidase towards substituted benzaldehydes. Acta Biochim Polon.; 51: 649-663.
• Panoutsopoulos GI, Kouretas D, Gounaris EG, Beedham C. (2004a) Metabolism of 2-phenylethylamine and phenylacetaldehyde by precision-cut guinea pig fresh liver slices. Eur J Drug Metab Pharmacokinet.; 29: 111-8.
• Panoutsopoulos GI, Kouretas D, Beedham C. (2004b) Contribution of aldehyde oxidase, xanthine oxidase and aldehyde dehydrogenase on the oxidation of aromatic aldehydes. Chem Res Toxicol.; 17: 1368-76.
• Pelsy G, Klibanov AM. (1983) Remarkable positional (regio) specificity of xanthine oxidase and some dehydrogenases in the reactions with substituted benzaldehydes. Biochim Biophys Acta.; 742: 352-7.
• Peterson GM, Boyle RR, Francis HW, Oliver NWJ, Paterson J, von Witt RJ, Taylor GR. (1990) Dosage prescribing and plasma oxipurinol levels in patients receiving allopurinol therapy. Eur J Clin Pharmacol.; 39: 419-21.
• Price RJ, Renwick AB, Walters DG, Young PJ, Lake BG. (2004) Metabolism of nicotine and induction of CYP1A forms in precision-cut rat liver and lung slices. Toxicol In Vitro.; 18: 179-85.
• Rajagopalan KV, Handler P. (1964) Hepatic aldehyde oxidase. II. Differential inhibition of electron transfer to various electron acceptors. J Biol Chem.; 239: 2022-6.
• Sasaki K, Hosoya R, Wang YM, Raulston GL. (1983) Formation and disposition of 7-hydroxymethotrexate in rabbits. Biochem Pharmacol.; 32: 503-7.
• Sipes IG, Fisher RL, Smith PF, Stine ER, Gandolfi AJ, Brendel K. (1987) A dynamic liver culture system: a tool for studying chemical biotransformation and toxicity. Arch Toxicol Supp.; 11: 20-33.
• Smith PF, Krack G, McKee RL, Johnson DG, Gandolfi AJ, Hruby VJ, Krumdieck CL, Brendel K. (1986) Maintenance of adult rat liver slices in dynamic organ culture. In Vitro Cell Dev Biol.; 22: 706-12.
• Stubley C, Stell JGP, Mathieson DW. (1979) The oxidation of azahetero-cycles with mammalian liver aldehyde oxidase. Xenobiotica.; 9: 475-84.
• Toutain HJ, Moronvalle HV, Sarsat JP, Chelin C, Hoet D, Leroy D. (1998) Morphological and functional integrity of precision-cut liver slices in rotating organ culture and multiwell plate culture: effects of oxygen uptake. Cell Biol Toxicol.; 14: 175-90.