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Design, Synthesis, and Biological Evaluation of Doxorubicin−Formaldehyde Conjugates Targeted to Breast Cancer Cells

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Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309-0215
Cite this: J. Med. Chem. 2004, 47, 5, 1193–1206
Publication Date (Web):January 24, 2004
Copyright © 2004 American Chemical Society

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    The anthracycline antitumor drug doxorubicin (DOX) has been utilized for decades as a broad-spectrum chemotherapeutic. Recent literature evidence documents the role of formaldehyde in the cytotoxic mechanism, and anthracycline−formaldehyde conjugates possess substantially enhanced activity in vitro and in vivo. Targeting a doxorubicin−formaldehyde conjugate specifically to cancer cells may provide a more efficacious chemotherapeutic. The design and 11-step synthesis of doxorubicin−formaldehyde conjugates targeted to the estrogen receptor, which is commonly overexpressed in breast cancer cells, are reported. The formaldehyde is incorporated in a masked form as an N-Mannich linkage between doxorubicin and salicylamide. The salicylamide triggering molecule, previously developed to release the doxorubicin−formaldehyde active metabolite, is tethered via derivatized ethylene glycols to an E and Z mixture of 4-hydroxytamoxifen. The targeting group, E/Z-4-hydroxytamoxifen, was selected for its ability to tightly bind the estrogen receptor and antiestrogen binding sites. The targeted doxorubicin−formaldehyde conjugates' estrogen receptor binding and in vitro growth inhibition were evaluated as a function of tether length. The lead compound, DOX-TEG-TAM, bearing a triethylene glycol tether, binds the estrogen receptor with a binding affinity of 2.5% relative to E/Z-4-hydroxytamoxifen and inhibits the growth of four breast cancer cell lines with 4-fold up to 140-fold enhanced activity relative to doxorubicin.

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    1H NMR spectra for all isolated intermediates and final products used in the biological evaluation; HPLC data for establishing degree of purity. This material is available free of charge via the Internet at

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