Induction of Apoptosis by 2-Chloro-2'deoxyadenosine (2-CdA) Alone and In Combination with other Cytotoxic Drugs: Synergistic Effects on Normal and Neoplastic Lymphocytes by Addition of Doxorubicin and Mitoxantrone

Abstract

2-CdA is active as a single agent in the treatment of low-grade lymphomas. We analyzed the induction of apoptosis by 2-CdA alone (n=5) and in combination with other drugs in peripheral lymphocytes from 25 patients with leukemic low-grade lymphomas and from 25 healthy volunteers. 2-CdA was tested in 4 escalating concentrations (0.05μg/ml to 0.4μg/ml). Linear regressions showed a dose dependant apoptosis rate of 0.29 × μg 2-CdA/mI + 0.11 (r²=0.88, p=0.006) in normal cells and 0.41 × μg 2-CdA/ml + 0.15 (r²=0.88, p=0.005) in leukemic cells. Intracellular metabolization of 2-CdA into 2-CdA-5'mono-, -di- and the active metabolite -triphosphate was analyzed by HPLC and paralleled the dose dependent increase of apoptosis. The combination of 2-CdA with doxorubicin or mitoxantrone had a synergistic effect on the induction of apoptosis (p<0.00l) in both normal and neoplastic lymphocytes, whereas 2-CdA plus etoposide or cytosine arabinoside were only additive. Due to the flat slope of the dose response of 2-CdA concentration on apoptosis we assume that higher in vivo dosages of 2-CdA in the treatment of low-grade lymphomas may not result in a higher clinical efficacy. The synergistic lymphocytotoxic effect of 2-CdA combined with doxorubicin or mitoxantrone may be relevant for new treatment approaches.

Key Words:

• apoptosis
• 2-CdA
• chronic lymphocytic leukemia
• low-grade lymphoma. HPLC
• flowcytom-etry