Highly purified isozymes of cytochrome P-450 catalyzed the formation of 3-glutathion-S-ylacetaminophen (GS-APAP) and 3-hydroxyacetaminophen (3-OH-APAP) from acetaminophen (APAP). A major isozyme from untreated male rats (P-450UT-A) catalyzed the formation of ca. 2.0 nmol/nmol of P-450/10 min of 3-OH-APAP and approximately 7.2 nmol of GS-APAP/nmol of P-450/10 min. Antibodies specific for cytochrome P-450UT-A caused a decrease in the amounts of both metabolites produced in microsomal incubations. In contrast to these results, two other constitutive P-450 isozymes from rat liver, cytochrome P-450UT-F and the female specific isozyme P-450UT-I, produced less of both oxidative metabolites. Moreover, they produced significantly more of the catechol metabolite than the glutathione conjugate. These results are in accord with the observation that male rats are more susceptible to acetaminophen hepatotoxicity than female rats. Isozymes induced by phenobarbital also produced more of the catechol than the glutathione conjugate. Conversely, the major isozyme induced by beta-naphthoflavone, cytochrome P-450 beta NF-B, produced a significantly greater amount of GS-APAP than 3-OH-APAP. When comparison was made to a major phenobarbital inducible form (cytochrome P-450PB-B) a definite isozyme specificity for the formation of the two metabolites was seen. The catechol was formed at rates of 2.21 and 0.53 nmol/nmol of P-450/10 min by cytochromes P-450PB-B and P-450 beta NF-B, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

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The agricultural fungicide, N-(3,5-dichlorophenyl)succinimide (NDPS), was shown to be nephrotoxic in rats. Previous studies have indicated that the metabolism of NDPS contributes to its mechanism of toxicity and both phase I and phase II pathways may be involved. In the current report, we investigated the detailed biotransformation of [(14)C]NDPS in rats using HPLC-ESI-MS. The chemical reactivity of the phase II NDPS metabolites was also evaluated. In vivo studies were conducted by administering [(14)C]NDPS to male Fischer 344 rats. Urine, tissue (liver and kidney), and plasma samples were analyzed. The mechanism of formation and chemical reactivity of the glucuronide and sulfate metabolites of NDPS were investigated in vitro using liver subcellular preparations. Major in vivo metabolites of NDPS were identified as the oxidative [N-(3,5-dichlorophenyl)-2- and 3-hydroxysuccinamic acid, 2-/3-NDHSA] and hydrolytic products [N-(3,5-dichlorophenyl)succinamic acid]. N-Acetylcysteine and cysteine (with intramolecular aminolysis) conjugates were also detected in rat urine and fecal extracts, respectively, suggesting the formation of reactive intermediate(s) in the metabolism of NDPS. Small amounts of the alcohol-O-glucuronide and O-sulfate of 2-/3-NDHSA were detected in rat urine, plasma, and tissue homogenates. The formation of these phase II metabolites was found to be mediated through the initial conjugation of N-(3,5-dichlorophenyl)-2-hydroxysuccinimide (NDHS) followed by hydrolysis. As compared to NDHS, NDHS-O-sulfate is approximately 500-fold more reactive toward GSH conjugation. In rat liver S9, fortifying phase II cofactors (UDPGA or PAPS) in incubation mixtures with NDHS also significantly increased the amount of GSH adducts produced. Results of this research demonstrate that phase II metabolites of NDPS were produced in rats. The formation of the alkyl alcohol-O-glucuronide and O-sulfate conjugates represents bioactivation pathways in the metabolism of NDPS that could potentially contribute to its mechanism of nephrotoxicity.

1. The thiazolidinedione (TZD) ring present in drugs available for type II diabetes may contribute to hepatic injury. Another TZD ring-containing compound, 3-(3,5dichlorophenyl)-2,4-thiazolidinedione (DCPT), produces liver damage in rats. Accordingly, the effects of gender, dose and time on DCPT hepatotoxicity were therefore evaluated. 2. Male rats were more sensitive to DCPT (0.4-1.0 mmol/kg by i.p. administration) as shown by increased serum alanine aminotransferase (ALT) levels and altered hepatic morphology 24 h post-dosing. Effects in both genders were dose-dependent. In males, DCPT (0.6 mmol/kg) produced elevations in ALTs and changes in liver sections 3 h after dosing that progressively worsened up to 12 h. DCPT-induced renal effects were mild. 3. We conclude that male rats are more susceptible to DCPT hepatotoxicity and that damage occurs rapidly. DCPT primarily affects the liver and may be a useful compound to investigate the role of the TZD ring in hepatic injury. However, the gender dependency and rapid onset of DCPT hepatotoxicity require further investigation.