Preparation of oligodeoxynucleotides containing a diastereoisomer of α-(N²-2′-deoxyguanosinyl)tamoxifen by phosphoramidite chemical synthesis

Women treated with an antiestrogen tamoxifen (TAM) for endocrine therapy or prevention of breast cancer show an increased risk of developing endometrial cancer. TAM-DNA adducts have been detected in the liver of rodents treated with TAM and in the endometrium of women taking TAM. The major TAM adducts have been identified as diastereoisomers of trans- and cis -forms of α-(N²-deoxyguanosinyl)tamoxifen (dG-N²-TAM) and α-(N²-deoxyguanosinyl)-N-desmethyltamoxifen. In the study presented here, we prepared oligodeoxynucleotides containing a diastereoisomer of dG-N²-TAM by phosphoramidite chemical synthesis. Initially, the trans- and cis-forms of α-aminotamoxifen (α-NH₂-TAM) were synthesized from α-hydroxytamoxifen using the Mitsunobu reaction followed by hydrolysis. Thereafter by coupling the trans- and cis-form of α-NH₂-TAM with the DMT-derivative of 2-fluoro-(O⁶-2-(trimethylsilyl)ethyl)-2′-deoxyinosine, the trans- and cis-forms of DMT-dG-N²-TAM, respectively, were prepared in high yield and used in the preparation of the phosphoramidite precursors. Large quantities of oligodeoxynucleotides containing a trans- or a cis-form of dG-N²-TAM were prepared efficiently by automated DNA synthesizer. The incorporation of dG-N²-TAM adduct into the oligodeoxynucleotides was confirmed using ³²P-postlabeling/polyacrylamide gel electrophoresis analysis. These site-specifically modified oligodeoxynucleotides will be used for exploring biological properties and three-dimensional structure of TAM - DNA adducts.