Further characterization of the DNA adducts formed in rat liver after the administration of tamoxifen, N-desmethyltamoxifen or N,N-didesmethyltamoxifen

determined by P-32-postlabelling, in the livers of rats given tamoxifen and the N-demethylated metabolites N-desmethyltamoxifen and N,N-didesmethyltam oxifen. Results show that after 4 days treatment (0.11 mmol/kg i.p,), simil ar levels of DNA damage were seen after treatment with either tamoxifen or N-desmethyltamoxifen [109 +/- 40 (n = 3) and 100 +/- 33 (n = 4) adducts/10( 8) nucleotides, respectively], even though the concentration of tamoxifen i n the livers of tamoxifen-treated rats was about half that of N-desmethylta moxifen in the N-desmethyltamoxifen-treated animals (51 +/- 16 and 100 +/- 8 nmol/g, respectively). Administration of N,N-didesmethyltamoxifen to rats resulted in a S-fold lower level of damage (19 adducts/10(8) nucleotides, n = 2), Following P-32-postlabelling and HPLC, hepatic DNA from rats treate d with tamoxifen and its metabolites showed distinctive patterns of adducts . Treatment of rats with N,N-didesmethyltamoxifen gave a major product that co-eluted with one of the minor adduct peaks seen in the livers of rats gi ven tamoxifen, Following dosing with N-desmethyltamoxifen, the major produc t co-eluted with one of the main peaks seen following treatment of rats wit h tamoxifen. This suggests that tamoxifen can be metabolically converted to N-desmethyltamoxifen prior to activation. However, analysis of the P-32-po stlabelled products from the reaction between alpha-acetoxytamoxifen and ca lf thymus DNA showed two main peaks, the smaller one of which (similar to 1 5% of the total) also co-eluted with that attributed to N-desmethyltamoxife n, This indicates that N-desmethyltamoxifen and N,N-didesmethyltamoxifen ar e activated in a similar manner to tamoxifen leading to a complex mixture o f adducts, Since an HPLC system does not exist that can fully separate all these 32P-postlabelled adducts, care has to be taken when interpreting resu lts and determining the relative importance of individual adducts and the m etabolites they are derived from in the carcinogenic process.