Human liver microsomal metabolism and DNA adduct formation of the tumorigenic pyrrolizidine alkaloid, riddelliine.
Riddelliine, a widespread naturally occurring genotoxic pyrrolizidine alkaloid, induced liver tumors in rats and mice in an NTP 2-year carcinogenicity bioassay. We have determined that riddelliine induces liver tumors in rats through a genotoxic mechanism involving the formation of (+/-)-6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP), which reacts with DNA to form a set of eight DNA adducts. To determine the relevance to humans of the results obtained in experimental animals, the metabolism of riddelliine was conducted using human liver microsomes. As with rat liver microsomes, DHP and riddelliine N-oxide were major metabolites in incubations conducted with human liver microsomes. The levels of DHP and riddelliine N-oxide were 0.20-0.62 and 0.03-0.15 nmol/min/mg protein, respectively, which are comparable to those obtained from rat liver microsomal metabolism. When metabolism was conducted in the presence of calf thymus DNA, the same set of eight DHP-derived DNA adducts was formed. Both the metabolism pattern and DNA adduct profile were very similar to those obtained from rat liver microsomes. When metabolism was conducted in the presence of the P450 3A4 enzyme inhibitor triacetyleandomycin, the formation of DHP and riddelliine N-oxide was reduced 84 and 92%, respectively. For DHP formation, the Km values were determined to be 0.37 +/- 0.05 and 0.66 +/- 0.08 mM from female rats and female humans; the Vmax values from female rat and human liver microsomal metabolism were 0.48 +/- 0.03 and 1.70 +/- 0.09 nmol/min/mg protein, respectively. These results strongly indicate the mechanistic data on liver tumor induction obtained for riddelliine in laboratory rodents is highly relevant to humans.
Xia
Qingsu
Q
National Center for Toxicological Research, Jefferson, Arkansas 72079, USA.
Chou
Ming W
MW
Kadlubar
Fred F
FF
Chan
Po-Cheun
PC
Fu
Peter P
PP
D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D002417:Cattle; D004247:DNA / Q000187:drug effects*; D018736:DNA Adducts / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000378:metabolism*; D008875:Middle Aged; D010759:Phosphorus Isotopes / Q000378:metabolism; D011763:Pyrrolizidine Alkaloids / Q000633:toxicity*; D051381:Rats; D011916:Rats, Inbred F344; D013045:Species Specificity; D014217:Troleandomycin / Q000494:pharmacology
D002273:Carcinogens; D018736:DNA Adducts; D004791:Enzyme Inhibitors; D010759:Phosphorus Isotopes; D011763:Pyrrolizidine Alkaloids; C013672:riddelliine; D004247:DNA; D014217:Troleandomycin