In vitro metabolism of two heterocyclic amines, 2-amino-9H-pyrido[2,3-b]indole (A(alpha)C) and 2-amino-3-methyl-9H-pyridol2,3-b]indole (MeA(alpha)C) in human and rat hepatic microsomes.
2-Amino-9H-pyrido[2,3-b]indole (A(alpha)C) and 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA(alpha)C) are two mutagenic and carcinogenic heterocyclic amines formed during ordinary cooking. In this study, we have investigated the in vitro metabolism of tritium-labelled A(alpha)C and MeA(alpha)C in hepatic microsomes from human pools, rats induced with polychlorinated biphenyl (PCB) (Aroclor 1254) and control rats. The microsomes were incubated with A(alpha)C and MeAaC and the detoxified and activated metabolites of A(alpha)C and MeA(alpha)C were separated and characterised by HPLC-MS. A(alpha)C is metabolised to two major and three minor detoxified metabolites, while MeA(alpha)C is metabolised to three major and one minor detoxified metabolites. Some A(alpha)C and MeA(alpha)C are activated by oxidation to the reactive metabolites N2-OH-A(alpha)C and N2-OH-MeA(alpha)C, respectively. These reactive N2-OH-metabolites react partially in the incubation system with formation of protein adducts, dimers and the parent compound by reduction of the N2-OH-metabolites. The distribution between the detoxified and activated metabolites in the different types of hepatic microsomes showed same pattern for both A(alpha)C and MeA(alpha)C. In PCB-induced rat microsomes, the major part of the metabolites are detoxified, only a little amount is activated. In control rat microsomes there is a fifty-fifty distribution between detoxification and activation, while the major part of the metabolites from the human microsomes are activated and reacts to form dimers and protein adducts. These data show that, in human hepatic microsomes compared to rat hepatic microsomes, a major part of A(alpha)C and MeA(alpha)C are metabolically activated to the reactive N2-OH-A(alpha)C and N2-OH-MeA(alpha)C.
Frederiksen
Hanne
H
Institute of Food Safety and Toxicology, Danish Veterinary and Food Administration, Søborg. hf@fdir.dk
Frandsen
Henrik
H
D000818:Animals; D002243:Carbolines / Q000138:chemical synthesis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D015394:Molecular Structure; D008970:Molecular Weight; D011078:Polychlorinated Biphenyls / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013057:Spectrum Analysis; D014316:Tritium
D002243:Carbolines; D014316:Tritium; C017821:2-amino-3-methyl-9H-pyrido(2,3-b)indole; D011078:Polychlorinated Biphenyls; C017822:2-amino-9H-pyrido(2,3-b)indole