title: Peroxidase activation of 4-hydroxytamoxifen to free radicals detected by EPR spectroscopy.
abstract: 4-Hydroxytamoxifen is a major metabolite of the antiestrogenic drug tamoxifen used in the treatment of women with breast cancer. 4-Hydroxytamoxifen is broken down by a horseradish peroxidase/H2O2 system very much more rapidly than tamoxifen and causes much greater DNA damage determined by 32P-postlabelling. EPR spin trapping of 4-hydroxytamoxifen reaction products in the presence of the free radical trap 5,5-dimethyl-1-pyrroline N-oxide, together with glutathione as a hydrogen donor, resulted in the generation of a species with the characteristics of the glutathione thiyl radical (aN approximately 15.3 G, aH approximately 16.2 G). Support for the creation of thiyl radicals comes from the close to stoichiometric time dependent formation of glutathione disulfide concomitant with the loss of glutathione. Similar results were obtained using 4-hydroxytoremifene but no radical formation or glutathione loss could be detected using 3-hydroxytamoxifen (droloxifene). On-line LC-ESI MS analysis of the incubation products from 4-hydroxytamoxifen has identified three products with a protonated molecular mass of 773, consistent with the formation of dimers of 4-hydroxytamoxifen. The role that radical mechanisms have in the carcinogenic effects of tamoxifen in the endometrium or other target organs of women taking this drug remains to be established.
authors.affiliation: MRC Toxicology Unit, Leicester, U.K.
authors.forename: A M
authors.initials: AM
authors.lastname: Davies
authors.forename: M E
authors.initials: ME
authors.lastname: Malone
authors.forename: E A
authors.initials: EA
authors.lastname: Martin
authors.forename: R M
authors.initials: RM
authors.lastname: Jones
authors.forename: R
authors.initials: R
authors.lastname: Jukes
authors.forename: C K
authors.initials: CK
authors.lastname: Lim
authors.forename: L L
authors.initials: LL
authors.lastname: Smith
authors.forename: I N
authors.initials: IN
authors.lastname: White
mesh_terms: D018736:DNA Adducts; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D046911:Macromolecular Substances; D010084:Oxidation-Reduction; D010761:Phosphorus Radioisotopes; D013113:Spin Labels; D013629:Tamoxifen / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D018736:DNA Adducts; D005609:Free Radicals; D046911:Macromolecular Substances; D010761:Phosphorus Radioisotopes; D013113:Spin Labels; D013629:Tamoxifen; C016601:afimoxifene; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D005978:Glutathione; D019803:Glutathione Disulfide