DNA adduct formation from quaternary benzo[c]phenanthridine alkaloids sanguinarine and chelerythrine as revealed by the 32P-postlabeling technique.
Using the 32P-postlabeling assay, we investigated the ability of quaternary benzo[c]phenanthridine alkaloids, sanguinarine, chelerythrine and fagaronine, to form DNA adducts in vitro. Two enhanced versions of the assay (enrichment by nuclease P1 and 1-butanol extraction) were utilized in the study. Hepatic microsomes of rats pre-treated with beta-naphthoflavone or those of uninduced rats, used as metabolic activators, were incubated in the presence of calf thymus DNA and the alkaloids, with NADPH used as a cofactor. Under these conditions sanguinarine and chelerythrine, but not fagaronine, formed DNA adducts detectable by 32P-postlabeling. DNA adduct formation by both alkaloids was found to be concentration dependent. When analyzing different atomic and bond indices of the C11-C12 bond (ring B) in alkaloid molecules we found that fagaronine behaved differently from sanguinarine and chelerythrine. While sanguinarine and chelerythrine showed a preference for electrophilic attack indicating higher potential to be activated by cytochrome P450, fagaronine exhibited a tendency for nucleophilic attack. Our results demonstrate that sanguinarine and chelerythrine are metabolized by hepatic microsomes to species, which generate DNA adducts.
Department of Biochemistry, Faculty of Science, Charles University, Albertov 2030, 128 40 2 Prague, Czech Republic. stiborov@natur.cuni.cz
Marie
M
Stiborová
Vilím
V
Simánek
Eva
E
Frei
Pavel
P
Hobza
Jitka
J
Ulrichová
D000470:Alkaloids / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism* / Q000494:pharmacology; D053119:Benzophenanthridines; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D018736:DNA Adducts / Q000096:biosynthesis*; D007364:Intercalating Agents / Q000378:metabolism* / Q000494:pharmacology; D007546:Isoquinolines; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D010617:Phenanthridines / Q000378:metabolism* / Q000494:pharmacology; D010761:Phosphorus Radioisotopes / Q000032:analysis; D011789:Quantum Theory; D051381:Rats; D017207:Rats, Sprague-Dawley
D000470:Alkaloids; D000972:Antineoplastic Agents, Phytogenic; D053119:Benzophenanthridines; D018736:DNA Adducts; D007364:Intercalating Agents; D007546:Isoquinolines; D010617:Phenanthridines; D010761:Phosphorus Radioisotopes; C002066:fagaronine; D003577:Cytochrome P-450 Enzyme System; C005705:sanguinarine; C016299:chelerythrine