Modulation of daunorubicin toxicity by liposomal encapsulation and use of specific inhibitors in vitro

Anthracyclines serve as a valuable tool in chemotherapy, but their usefulness is often limited by the occurrence of resistance mechanisms in tumor cells. Resistance of tumor cells is a multifactorial event, where several mechanisms act concurrently, including drug efflux and enzymatic drug inactivation. Liposomal encapsulation of anthracyclines has been discussed as a successful regimen to overcome drug resistance. Our investigations were carried out on a daunorubicin (DRC) sensitive breast cancer cell line and two DRC resistant sublines generated thereof. In all three cell lines, the extent of DRC detoxification via carbonyl reduction to daunorubicinol (DRCOL) was determined. In addition, rutin, the most effective inhibitor of carbonyl reducing enzymes, was tested to affect DRCOL formation. DRC IC(50) values were determined in relation to several combinations of DRC administration, (a) liposomal encapsulated DRC, (b) addition of verapamil (inhibitor of drug efflux), (c) addition of rutin (inhibitor of DRC carbonyl reduction). We could show that DRC sensitive and resistant breast cancer cell lines are able to catalyze DRC detoxification via carbonyl reduction to DRCOL. Rutin was shown to inhibit this reaction, but could not serve as an enhancer of DRC toxicity in MTT tests. Verapamil was effective only in resistant cells due to the overexpression of P-glycoprotein 170. Liposomal encapsulation of DRC did not show the expected increase in DRC toxicity in the present tumor cell model.