Evaluation of doxorubicin in combination with mafosfamide for in vitro elimination of myeloid and lymphoid tumor cells from human bone marrow

Abstract

In an attempt to improve in vitro pharmacological purging of autologous grafts, the ability of doxorubicin (DOX), alone and in combination with mafosfamide (AZ), to eliminate tumor cells from human bone marrow was assessed. HL60 and Raji cells were mixed with a 20-fold excess of normal marrow cells and were incubated either with DOX (0.4-3.2 micrograms/ml) for 1 h or AZ (20-140 micrograms/ml) for 30 min or both drugs sequentially. Cytotoxicity was evaluated on tumor cells and GM-CFU by clonogenic assays and on earlier hemopoietic progenitors by liquid long-term marrow cultures (LTMC) for 5 weeks. DOX at 3.2 micrograms/ml and AZ at 140 micrograms/ml spared 1.08 and 1.23% of GM-CFU respectively, and yielded similar tumor cell log-kills for HL60 cells (3.04 and 2.95) and Raji cells (3.24 and 3.40). With the combination of AZ and DOX, the best therapeutic index was observed when the cells were incubated with AZ prior to DOX. Under these conditions, AZ at 80 micrograms/ml together with DOX at 1.6 micrograms/ml significantly increased log-kill values for HL60 cells to 3.96 by a synergistic effect and for Raji cells to 3.85 by an additive effect. In LTMC, GM-CFU recovery after treatment with AZ alone and with the combination of AZ and DOX was 59.9 and 20.0%, respectively, while it was 7.9 and 2.9% at culture initiation. These results suggest that the purging efficiency of DOX is comparable to that of AZ and may be enhanced by combination with AZ.