The natural product betulin is under investigation for several therapeutic indications, however little is known about its metabolism. In the present study, the glucuronidation and sulfation of betulin in human and rat liver microsomes and cytosol were tested. We further identified the main UDP-glucuronosyltransferases (UGTs) and sulfotransferases (SULTs) involved in these two metabolism pathways. Results showed that one betulin glucuronide metabolite was observed after incubation with human and rat liver microsomes. The glucuronidation of betulin in human liver microsomes had a Km value of 21.1 ± 5.93 μM and a Vmax value of 6.39 ± 0.66 pmol/min/mg protein. The glucuronidation activity in rats was too low to get enzyme kinetic parameters. Among the 11 recombinant UGT enzymes investigated, UGT1A3 and UGT1A4 were identified as the major enzymes catalyzing the glucuronidation of betulin [Km values of 10.12 ± 8.09 and 8.04 ± 3.96 μM, Vmax values of 6.71 ± 1.51 and 5.98 ± 0.76 nmol/min/(mg protein)]. Two betulin sulfate metabolites were found in human and rat liver cytosols. Human and rat liver had similar affinity for the formation of these two metabolites, the apparent Vmax for betulin sulfate I was higher than that for betulin sulfate II in both species. Among the SULT isoforms studied, SULT2A1 was the major isoenzyme involved in the betulin sulfation metabolism in human liver cytosol. The results suggest that glucuronidation and sulfation are important metabolism pathways for betulin, and UGT1A3, UGT1A4 and SULT2A1 play the major roles in betulin glucuronidation and sulfation.
Keywords: Betulin; Glucuronidation; Isoenzymes; Sulfation.
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