Hepatobiliary transport of diflunisal conjugates and taurocholate by the perfused rat liver: The effect of chronic exposure of rats to diflunisal

Acyl glucuronides are reactive electrophilic metabolites of carboxylate drugs which can form covalent adducts with endogenous macromolecules such as serum albumin and hepatic proteins. Such adducts have been suggested as initiating factors in certain immune and toxic responses to acidic drugs. In the present study, pretreatment of rats with high daily doses (50 mg/kg orally) of the nonsteroidal anti-inflammatory drug (NSAID) diflunisal (DF) for 35 days, followed by perfusion of the isolated liver with 3 mg DF for 3 hr, resulted in appreciable concentrations of covalent adducts of DF with hepatic tissue (3.68 μg DF/g liver). Immunoblotting using a rabbit polyclonal DF antiserum showed the major DF-modified bands at about 110, 140 and 200 kDa. A vehicle-pretreated control group achieved adduct concentrations of only 0.37 μg DF/g liver, with the 200 kDa band not detectable in immunoblots. Elimination of DF from perfusate of the isolated perfused rat liver (IPRL) preparation was the same (t( 1/4 ) about 3.4 hr) in both DF and vehicle-pretreated groups. Appearance of the sulfate (DS) conjugate, the major metabolite in perfusate, was also similar. However, higher concentrations of the acyl glucuronide (DAG) and phenolic glucuronide (DPG) conjugates were found in perfusate at later times, though a statistically significant difference in area under the concentration-time curve was found only in the case of DAG. At 3 hr, recoveries of dose as DAG and DPG were significantly higher in perfusate, but not in bile. No significant differences in uptake and biliary excretion of taurocholate were found between the two groups. The finding of higher perfusate concentrations of DAG and DPG could signal a minor compromise to biliary excretion processes for the glucuronides, though whether such a result is simply coincident with or attributable to DAG-derived covalent DF-protein adducts in liver remains indeterminate