Etienne Benoit

INDUCING EFFECT OF ALBENDAZOLE ON RAT LIVER DRUG-METABOLIZING ENZYMES AND METABOLITE PHARMACOKINETICS

Albendazole (ABZ), methyl (5-(propylthio)-1H-benzimidazol-2-yl)carbamate, is a broad spectrum anthelmintic drug. S-oxidation to the sulfoxide (SO-ABZ) and the sulfone (SO2-ABZ) are the first steps of its bioconversion. SO-ABZ is pharmacologically active and embryotoxic in rats. In the present study, rat liver microsomal drug-metabolizing enzymes were assayed after 10 days oral administration with 40 mumol ABZ/kg per day. The activities of 4-nitroanisole O-demethylase, benzo[a]pyrene hydroxylase, 7-ethoxycoumarin O-deethylase, and 7-ethoxyresorufin O-deethylase increased 6-, 7-, 8-, and 30-fold, respectively. By immunoblotting an increase in cytochrome P-448 was observed. UDP-glucuronosyltransferase (GT) type 1 activities (1-naphthol, 7-hydroxycoumarin, 4-nitrophenol, and 4-methylumbelliferone) were significantly higher than in control microsomes (3- to 4-fold), while GT type 2 activities and bilirubin-GT remained unchanged. Microsomal epoxide hydrolase (benzo[a]pyrene oxide) increased 2-fold. Microsomal gamma-glutamyltransferase activity was unchanged. The in vivo SO-ABZ plasma level was decreased when the SO2-ABZ plasma level was increased. In vitro sulfoxidation and sulfonation were, however, unchanged. Although a range of imidazole derivatives, including benzimidazole itself, were commonly reported as inhibitors of monooxygenase activities, ABZ behaved as an inducer of cytochrome P-448, GT1, and epoxide hydrolase.

Subclinical doses of T-2 toxin impair acquired immune response and liver cytochrome P450 in pigs

This study was designed to investigate the effect of subclinical doses of T-2 toxin on liver drug-metabolizing enzymes and the immune response. Pigs were offered over a 28-day period either a control diet or diets contaminated with 540, 1324 or 2102microg pure T-2toxin/kg feed. Pigs were immunized with ovalbumin and subsequent humoral and cellular immune responses measured. Monooxygenase and transferase enzyme activities and protein expression were investigated in liver tissue samples. Pigs fed 1324 or 2102microg T-2toxin/kg feed exhibited reduced anti-ovalbumin antibody production without significant alteration to specific lymphocyte proliferation. The livers of pigs exposed to T-2 toxin presented normal cytochrome P450 content, UGT 1A and P450 2B, 2C or 3A protein expression, and glutathione- and UDP glucuronosyl-transferase activities. However, P450 1A related activities (ethoxyresorufin O-deethylation and benzo-(a)-pyrene hydroxylation) were reduced for all pigs given T-2 toxin, with P450 1A protein expression decreased in pigs fed the highest dose. In addition T-2 toxin exposure reduced certain N-demethylase activities. The results of this study confirm the immunotoxic properties of T-2 toxin, in particular toward the humoral immune response. The reduction of monooxygenase activities, even though the liver presented no tissue lesion or lipid peroxidation, suggests possible deleterious interactions of T-2 toxin with these enzymes.

Adrenal cortical response in clinically normal dogs before and after adaptation to a housing environment

58 dogs (29 males and 29 females) selected as healthy on clinical and biochemical evaluations were subjected to an ACTH adrenal function test 2 days after their admission to a veterinary hospital (t+0). Basal female serum cortisol concentrations were significantly higher than concentrations in males (77 nmol/l versus 43 nmol/l; P<0·01). Concentrations post stimulation were not statistically different (P>0·05) between males and females: 306 (±69) nmol/l versus 291 (±73) nmol/l, respectively. Twelve dogs (6 males and 6 females), randomly selected from the 58, were subjected to the same test 5 weeks later (t+5) and 12 weeks later (t+12). Basal cortisol concentrations were lower at t+5 or at t+12 than at t+0. Post stimulation mean cortisol concentrations were lower in males than in females at t+5 (162 versus 232 nmol/l; P<0·05) but not at t+0 (262 versus 320 nmol/l; P>0·5) and t+12 (188 versus 233 nmol/l; P>0·05). These findings are indicating an increased susceptibility of bitches to environmental stress.

Physiological factors affecting the expression of FMO1 and FMO3 in the rat liver and kidney.

FMO1 and FMO3, the main FMOs described in the rat, are highly expressed in the liver and the kidney. The age, from 3 to 11 weeks, and gender-dependent expression of FMO1 and FMO3 in the rat liver and kidney were investigated. Based on the enzyme activities, protein levels and mRNA levels, this study demonstrates an important increase in the expression of the FMO3 in the liver of male rats during a period that corresponds to the acquisition of the sexual maturity. Rat liver FMO1 remains unchanged during this period of observation. The evolutions of both isoforms in the kidney of the male rat are similar to those observed in the liver. On the contrary, the important decrease in the total flavin-containing monooxygenase (FMO) activity observed in the liver of female rat is linked to a considerable decrease in the FMO1-dependent activity, FMO1 protein and FMO1 mRNA levels as a function of age. The expression of the FMO3 in the liver does not seem to be affected by the age of the female rat. Inversely, the expression of FMO1 in the female rat kidneys does not seem to be modified as a function of age while the expression of FMO3 is strongly increased.

Dietary glucomannan improves the vaccinal response in pigs exposed to aflatoxin B1 or T-2 toxin

The aim of the study was to investigate whether dietary supplementation with yeast-derived glucomannan protects pigs against the deleterious effects that exposure to aflatoxin B1 (AFB1) or T-2 toxin has on the vaccinal immune response and drug-metabolising enzymes. Three doses of pure mycotoxin (AFB1 trial: 482, 968 and 1,912 µg/kg feed; T-2 toxin trial: 593, 1,155 and 2,067 µg/kg feed) with or without dietary glucomannan supplementation (2 g/ kg feed) were tested in weaned pigs for 28 days. At days 4 and 15 pigs were immunised with ovalbumin to study the humoral and cell-mediated antigen-specific immune responses. The effects of AFB1 and T-2 toxin intake alone in pigs have already been published. In all parameters investigated no differences were apparent between animals receiving the unsupplemented control diet or the control diet containing glucomannan. In the AFB1 trial glucomannan decreased the severity of liver lesions in animals exposed to 968 µg/kg feed. Exposure to both AFB1 and T-2 toxin were as...

(−)-R-fenoprofen: Formation of fenoprofenyl-coenzyme A by rat liver microsomes

The thioesterification of fenoprofen (FPF) by rat liver microsomes has been studied using an HPLC method enabling direct quantification of the FPF-CoA produced. Over the concentration range studied (5-400 microM), studies showed the participation of a single CoA ligase in the formation of FPF-CoA, in contrast with the involvement of several isozymes with different affinities, that has been found with ibuprofen (IPF). The Km for the reaction was dependent upon the presence of non-ionic detergent, a concentration of 0.05% Triton X-100 reducing the Km from 397 to 20 microM although the detergent had no effect on Vmax. The microsomal long-chain fatty acid CoA ligase was markedly enantioselective towards (-)-R-FPF and the formation of (-)-R-FP-CoA was inhibited by both the (+)-S enantiomer and palmitic acid.

Stereoselective S-oxygenation of an aryl-trifluoromethyl sulfoxide to the corresponding sulfone by rat liver cytochromes P450

Toltrazuril sulfoxide (TZR.SO) is the metabolite of the antiparasitic drug toltrazuril (TZR; 1-methyl-3-[3-methyl-4-[4-[trifluoromethyl]thio]phenoxy]phenyl- 1,3,5-triazine-2,4,6(1H,3H,5H)-trione). The results of the present paper demonstrate that TZR.SO was metabolized by rat liver microsomes to the corresponding sulfone (TZR.SO2). The reaction was mediated almost exclusively by different cytochromes P450, the most active being cytochromes P450 3A. TZR.SO exists as a racemic mixture; when each enantiomer was incubated separately in the presence of untreated rat liver microsomes, a 7.3-fold difference in the rate of S-oxygenation was found, indicating a marked substrate enantioselectivity for the reaction.

KINETICS OF FOUR METABOLITES OF FEBANTEL IN COW'S MILK

The mammary elimination of four potentially toxic metabolites of Febantel has been investigated by HPLC methods in the cow after the administration of a 7,5 mg/kg single oral experimental dose of this anthelmintic.The total of the four metabolites is equal to about 0.4 ppm 12 hours after the treatment. The individual levels fall down under the detection limit between the third and the sixth milking.