Metabolism of the Food-Associated Carcinogen 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by Human Intestinal Microbiota

Lynn Vanhaecke, Nathalie Van Hoof, Willem Van Brabandt, Bram Soenen, Arne Heyerick, Norbert De Kimpe, Denis De Keukeleire, Willy Verstraete, Tom Van de Wiele

2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine is a putative human carcinogenic heterocyclic aromatic amine formed from meat and fish during cooking. Although the formation of hazardous PhIP metabolites by mammalian enzymes is well-documented, nothing is known about the PhIP transformation potency of human intestinal bacteria. In this study, the in vitro metabolism of PhIP by human fecal samples was investigated. Following anaerobic incubation of PhIP with stools freshly collected from six healthy volunteers, we found that PhIP was extensively transformed by the human intestinal bacteria. HPLC analysis showed that the six human fecal microbiota transformed PhIP with efficiencies from 47 to 95% after 72 h incubation, resulting in one major derivative. ESI-MS/MS, HRMS, 1D (1H, 13C, DEPT) and 2D (gCOSY, gTOCSY, gHMBC, gHSQC) NMR, and IC analysis elucidated the complete chemical identity of the microbial PhIP metabolite as 7-hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3',2':4,5]imidazo[1,2-a]pyrimidin-5-ium chloride. At present, no information is available about the biological activity of this newly discovered bacterial PhIP metabolite. Our findings however suggest that bacteria derived from the human intestine play a key role in the activation or detoxification of PhIP, a digestive fate ignored so far in risk assessments. Moreover, the variation in transformation efficiency between the human microbiota indicates interindividual differences in the ability to convert PhIP. This may predict individual susceptibility to carcinogenic risk from this suspected dietary carcinogen.