Transferrin conjugates of adriamycin are cytotoxic without intercalating nuclear DNA.

Studies of the biological chemistry of most anticancer drugs have revealed their cytotoxicity is expressed after the drugs have entered cells. It is thought that anthracycline antitumor drugs exert their cytotoxicity by entering cells, diffusing into nuclei, and inhibiting topoisomerase II and/or intercalating DNA base pairs. In order to deliver anthracyclines to transferrin (TRF) receptors on the plasma membranes of human tumor cells, we have prepared conjugates of adriamycin (ADR) with human TRF. These TRF-ADR conjugates were found to be stable at low pH and to exert more efficient cytotoxicity than free drug. By using spectrofluorometry, we found that the fluorescence of ADR within the conjugate was quenched by native DNA, demonstrating the presence of conformationally available drug to intercalate with nuclear DNA. However, fluorescence was not quenched when conjugate was reacted with viable cells, indicating that ADR did not reach the nucleus. Results of fluorescence microscopy experiments confirmed that free but not conjugated ADR reached the nuclei of viable cells, and TRF-ADR conjugates labeled with fluorescein isothiocyanate were found to initiate lateral diffusion as determined by patch and cap reactions. The involvement of TRF receptors was shown by flow cytometry experiments in which native TRF inhibited binding of fluorescein-labeled TRF-ADR conjugates. These data suggest that TRF-ADR conjugates mediate cytotoxicity by a mechanism other than intercalation with nuclear DNA. This mechanism, revealed by conjugating ADR to a TRF carrier, may not initiate complications such as cardiotoxicity and drug resistance.