Hepatic modeling of metabolite kinetics in sequential and parallel pathways: salicylamide and gentisamide metabolism in perfused rat liver.
Previous data on salicylamide (SAM) metabolism in the perfused rat liver had indicated that SAM was metabolized by three parallel (competing) pathways: sulfation, glucuronidation, and hydroxylation, whereas sequential metabolism of the hydroxylated metabolite, gentisamide (GAM), was solely via 5-glucuronidation to form GAM-5G. However, under comparable conditions, preformed GAM formed mainly two monosulfate conjugates at the 2- and 5-positions (GAM-2S and GAM-5S); 5-glucuronidation was a minor pathway. In the present study, the techniques of normal (N) and retrograde (R) rat liver perfusion with SAM and mathematic modeling on SAM and GAM metabolism were used to explore the role of enzymic distributions in determining the dissimilar fates of GAM, as a generated metabolite of SAM or as preformed GAM. Changes in the steady-state extraction ratio of SAM (E) and metabolite formation ratios between N and R perfusions were used as indices of the uneven distribution of enzyme activities. Two SAM concentrations (134 and 295 microM) were used for single-pass perfusion: the lower SAM concentration exceeded the apparent Km for SAM sulfation but was less than those for SAM glucuronidation and hydroxylation; the higher concentration exceeded the apparent Km's for SAM sulfation and glucuronidation but was less than the Km for hydroxylations. Simulation of SAM metabolism data was carried out with various enzyme distribution patterns and extended to include GAM metabolism. At both input concentrations, E was high (0.94 at 134 microM and 0.7 at 295 microM) and unchanged during N and R, with SAM-sulfate (SAM-S) as the major metabolite and GAM-5G as the only detectable metabolite of GAM. Saturation of SAM sulfation occurred at the higher input SAM concentration as shown by a decrease in E and a proportionally less increase in sulfation rates and proportionally more than expected increases in SAM hydroxylation and glucuronidation rates. At both SAM concentrations, the steady-state ratio of metabolite formation rates for SAM-S/SAM-G decreased when flow direction changed from N to R. An insignificant decrease in SAM-S/SAM-OH was observed at the low input SAM concentration, due to the small amount of SAM-OH formed and hence large variation in the ratio among the preparations, whereas at the high input SAM concentration, the decrease in SAM-S/SAM-OH with a change in flow direction from N to R was evident. The metabolite formation ratio, SAM-G/SAM-OH, however, was unchanged at both input concentrations and flow directions.(ABSTRACT TRUNCATED AT 400 WORDS)
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Faculty of Pharmacy, University of Toronto, Ontario, Canada.
Pang
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D000818:Animals; D001549:Benzamides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008722:Methods; D008954:Models, Biological; D010477:Perfusion; D010599:Pharmacokinetics; D051381:Rats; D011919:Rats, Inbred Strains; D012457:Salicylamides / Q000378:metabolism
D001549:Benzamides; D012457:Salicylamides; C055881:gentisamide; C031060:salicylamide