title: Differences in metabolite-mediated toxicity of tamoxifen in rodents versus humans elucidated with DNA/microsome electro-optical arrays and nanoreactors.
abstract: Tamoxifen, a therapeutic and chemopreventive breast cancer drug, was chosen as a model compound because of acknowledged species specific toxicity differences. Emerging approaches utilizing electro-optical arrays and nanoreactors based on DNA/microsome films were used to compare metabolite-mediated toxicity differences of tamoxifen in rodents versus humans. Hits triggered by liver enzyme metabolism were first provided by arrays utilizing a DNA damage end point. The arrays feature thin-film spots containing an electrochemiluminescent (ECL) ruthenium polymer ([Ru(bpy)(2)PVP(10)](2+); PVP, polyvinylpyridine), DNA, and liver microsomes. When DNA damage resulted from reactions with tamoxifen metabolites, it was detected by an increase in light from the oxidation of the damaged DNA by the ECL metallopolymer. The slope of ECL generation versus enzyme reaction time correlated with the rate of DNA damage. An approximate 2-fold greater ECL turnover rate was observed for spots with rat liver microsomes compared to that with human liver microsomes. These results were supported by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of reaction products using nanoreactors featuring analogous films on silica nanoparticles, allowing the direct measurement of the relative formation rate for alpha-(N(2)-deoxyguanosinyl)tamoxifen. We observed 2-5-fold more rapid formation rates for three major metabolites, i.e., alpha-hydroxytamoxifen, 4-hydroxytamoxifen, and tamoxifen N-oxide, catalyzed by rat liver microsomes compared to human liver microsomes. Comparable formation rates were observed for N-desmethyl tamoxifen with rat and human liver microsomes. A better detoxifying capacity for human liver microsomes than rat liver microsomes was confirmed utilizing glucuronyltransferase in microsomes together with UDP-glucuronic acid. Taken together, lower genotoxicity and higher detoxication rates presented by human liver microsomes correlate with the lower risk of tamoxifen in causing liver carcinoma in humans, provided the glucuronidation pathway is active.
authors.affiliation: Department of Chemistry, 55 N. Eagleville Road, University of Connecticut, Storrs, Connecticut 06269, USA.
authors.forename: Linlin
authors.initials: L
authors.lastname: Zhao
authors.forename: Sadagopan
authors.initials: S
authors.lastname: Krishnan
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.forename: John B
authors.initials: JB
authors.lastname: Schenkman
authors.forename: James F
authors.initials: JF
authors.lastname: Rusling
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000633:toxicity*; D004247:DNA / Q000737:chemistry; D018736:DNA Adducts / Q000032:analysis / Q000737:chemistry; D004249:DNA Damage; D055664:Electrochemical Techniques; D014453:Glucuronosyltransferase / Q000378:metabolism; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008862:Microsomes, Liver / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D036103:Nanotechnology; D055095:Optics and Photonics; D051381:Rats; D013045:Species Specificity; D013629:Tamoxifen / Q000378:metabolism / Q000633:toxicity*; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000970:Antineoplastic Agents; D018736:DNA Adducts; D013629:Tamoxifen; D004247:DNA; D014453:Glucuronosyltransferase