Glucuronidation of 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine by human microsomal UDP-glucuronosyltransferases: identification of specific UGT1A family isoforms involved
Sa. Nowell et al., Glucuronidation of 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine by human microsomal UDP-glucuronosyltransferases: identification of specific UGT1A family isoforms involved, CARCINOGENE, 20(6), 1999, pp. 1107-1114
2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine is a heterocyclic aromatic
amine found in cooked meats and dietary exposure to PhIP has been implicate
d in the etiology of colon cancer in humans. PhIP, along with other heteroc
yclic aromatic amines, requires metabolic activation to exhibit genotoxic e
ffects, PhIP is initially oxidized by the activity of cytochrome P4501A2 to
produce 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-OH-PhIP)
, a reaction occurring primarily in the liver. Whereas subsequent biotransf
ormation of N-OH-PhIP via acetylation or sulfation can produce reactive ele
ctrophiles that readily bind to DNA, N-glucuronidation, catalyzed by UDP-gl
ucuronosyltransferases (UGTs), functions as a detoxification mechanism. Alt
hough hepatic glucuronidation of N-OH-PhIP has been well characterized, the
extrahepatic metabolism of this compound is poorly understood. Studies in
our laboratory now indicate that the intestinal tract, and particularly the
colon, is a significant site of glucuronidation of N-OH-PhIP, When assays
were performed with microsomes prepared from the mucosa of the intestinal t
ract, it was determined that glucuronidation of N-OH-PhIP occurs throughout
the intestinal tract, with activity approximately three times higher in th
e colon as that found in the upper intestine. Glucuronidation rates from co
lon microsomes showed considerable interindividual variability and incubati
on with N-OH-PhIP yielded two glucuronides, HPLC analysis showed that the p
redominant product formed is the N-OH-PhIP-N-2-glucuronide, while the N3-gl
ucuronide accounts for <10% of the total glucuronidation product. These rat
es approach the rates found in human liver microsomes, demonstrating the si
gnificance of extrahepatic metabolism of this food-borne carcinogen. Subseq
uent assays with human recombinant UGTs demonstrated that at least four hum
an UGT isoforms, all from the UGT1A subfamily, are capable of catalyzing th
e biotransformation of N-OH-PhIP, Members of the UGT2B family available for
this study did not conjugate N-OH-PhIP, although immunoinhibition studies
in human liver microsomes strongly suggest the involvement of a UGT2B isofo
rm(s) in this organ.