Determination of DNA damage in F344 rats induced by geometric isomers of tamoxifen and analogues.
To investigate the activation mechanisms involved in tamoxifen carcinogenicity, analogues of tamoxifen isomers modified at the ethyl group were synthesized and assessed for their ability to induce hepatic DNA damage following their administration to female F344 rats. The cis isomer was prepared by acid-catalyzed isomerization of tamoxifen and isolated by preparative HPLC. The active metabolite alpha-hydroxytamoxifen and geometric isomers of bromotamoxifen and C-desmethylenetamoxifen, analogues in which the ethyl group has been replaced by a bromine atom and methyl group, respectively, were synthesized according to published procedures. The levels of hepatic DNA adducts induced were determined by 32P-postlabeling. Bromotamoxifen and tamoxifen 1,2-epoxide caused no detectable DNA damage relative to controls. Trans isomers of tamoxifen, C-desmethylenetamoxifen, and alpha-hydroxytamoxifen all produced DNA adducts at a 5-90-fold higher level than the corresponding cis isomers. In contrast, both the cis and trans isomers of alpha-hydroxytamoxifen showed similar reactivity toward calf thymus DNA in vitro. Molecular models of alpha-hydroxytamoxifen isomers suggest this difference in DNA adduct-forming ability is due to steric hindrance of the enzymes involved in the activation of this metabolite. There were high adduct levels in the liver, but no uterine DNA adducts were detected in rats treated with alpha-hydroxytamoxifen. This suggests that in contrast to the liver, alpha-hydroxytamoxifen is not further activated in rat uterus. This may help to explain the absence of uterine tumors in rats following long-term tamoxifen treatment.
Brown
K
K
Pharmaceutical Chemistry, University of Bradford, West Yorkshire BD7 1DP, U.K.
Brown
J E
JE
Martin
E A
EA
Smith
L L
LL
White
I N
IN
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification; D004249:DNA Damage / Q000187:drug effects*; D004965:Estrogen Antagonists / Q000737:chemistry / Q000633:toxicity*; D005260:Female; D005583:Fourier Analysis; D066298:In Vitro Techniques; D007536:Isomerism; D008099:Liver / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D008958:Models, Molecular; D051381:Rats; D011916:Rats, Inbred F344; D013629:Tamoxifen / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity*
D004965:Estrogen Antagonists; D013629:Tamoxifen; D004247:DNA