Synthesis and Characterization of Chemically Stable N7-dG Estrone and Catechol Adducts

Endogenous estrogens are implicated in carcinogenesis through both estrogen receptor-mediated cell proliferation and the direct genotoxicity of reactive metabolites. Oxidative metabolism of estrogens produces catechol estrogens that are further converted to electrophilic ortho-quinones capable of alkylating DNA. The prevailing model of mutagenesis proposes that these N3Ade and N7Gua adducts depurinate to form abasic sites that induce mutations initiating hormone-related cancers. However, the mutation spectrum observed in experimental data is inconsistent with this mechanism, and synthetic studies of estrogen-DNA adducts have relied on acidic conditions that artificially promote depurination, leaving stable N7-dG lesions poorly understood. To address this, we synthesized stable N7-dG catechol and estrone adducts using 2′-fluorinated deoxyguanosine, a modification that inhibits N-glycosidic bond cleavage. ROESY 2D NMR spectroscopy revealed through-space correlations consistent with a preferred anti-conformation in solution, supported by molecular modeling. Structural analysis suggests that these cationic aryl adducts likely preserve the Watson–Crick base pairing edge but may promote tautomerization capable of altering base pairing and generating G-to-A mutations. These findings provide the first synthesized stable models of N7-dG estrogen adducts and may support an alternative mechanism of estrogen-induced mutagenesis independent of depurination, enabling future biochemical investigations of related DNA repair and mutagenesis.