title: Comparison of the interaction of pyrazole and its metabolite 4-hydroxypyrazole with rat liver microsomes.
abstract: Pyrazole is known to interact with and to induce cytochrome P-450 IIE1. Since pyrazole is oxidized by rat liver microsomes to 4-hydroxypyrazole, and several of the actions of pyrazole have been ascribed to its metabolite, experiments were conducted to evaluate the interactions of 4-hydroxypyrazole with microsomes, and to compare these to pyrazole itself. Rats were injected with doses of 4-hydroxypyrazole ranging from 2 to 100 mg/kg body weight/day for 2 days. A slight increase of total cytochrome P-450 was observed at low doses, followed by a decrease at higher concentrations. NADPH-cytochrome P-450 reductase activity was not affected. The oxidation of aniline or dimethylnitrosamine was increased about 50% by the 4-hydroxypyrazole treatment; however, this extent of increase was much less than that produced by pyrazole treatment. In vitro, 4-hydroxypyrazole produced a type II binding spectrum with microsomes, with a peak at about 425 nm and a trough at about 395 nm. The affinity for 4-hydroxypyrazole was increased from a value of about 0.60 mM in control microsomes to a value of about 0.40 mM in microsomes from pyrazole-treated rats. These values are 2-fold greater than those observed with pyrazole as the ligand. 4-Hydroxypyrazole inhibited the microsomal oxidation of ethanol; kinetics of inhibition were mixed. The apparent KI for 4-hydroxypyrazole inhibition of ethanol oxidation by microsomes was about 4 mM, which is about an order of magnitude greater than that for pyrazole. The in vivo and in vitro interactions of 4-hydroxypyrazole with microsomes appear to be similar to those described for pyrazole; however, these interactions are considerably less effective than those of the parent drug, pyrazole. Thus, although some actions of pyrazole may be due to the metabolite 4-hydroxypyrazole, it appears that the induction of P-450 IIE1 and the in vitro interactions of pyrazole with microsomes is not likely to be mediated by prior metabolism of pyrazole to 4-hydroxypyrazole.
authors.affiliation: Department of Biochemistry, Mount Sinai School of Medicine, New York, NY 10029.
authors.forename: L A
authors.initials: LA
authors.lastname: Clejan
authors.forename: A I
authors.initials: AI
authors.lastname: Cederbaum
mesh_terms: D000815:Aniline Hydroxylase / Q000378:metabolism; D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis; D004790:Enzyme Induction / Q000187:drug effects; D000431:Ethanol / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010084:Oxidation-Reduction; D011720:Pyrazoles / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D011720:Pyrazoles; D000431:Ethanol; C014606:4-hydroxypyrazole; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D000815:Aniline Hydroxylase; D009251:NADPH-Ferrihemoprotein Reductase