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Complementary antineoplastic activity of the cytosine nucleoside analogues troxacitabine (Troxatyl) and cytarabine in human leukemia cells

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Abstract

Purpose

Troxacitabine (BCH-4556, l-(−)-OddC, Troxatyl) is a novel β-l-nucleoside analogue with potent antineoplastic activity both in vitro and in several tumor models in vivo, and is presently in phase II clinical trials. The combination of the cytosine analogues troxacitabine and araC (1-β-d-arabinofuranosylcytosine, cytarabine) has shown promising activity in patients with acute myelogenous leukemia. To further examine the interactions between these two analogues, we investigated the in vitro and in vivo effects of their combination against a human leukemia cell line, CCRF-CEM.

Methods

The in vitro cytotoxic effect of the combination of troxacitabine and araC on the survival of CCRF-CEM cells was measured using a standard MTT assay and combination indices were generated with the CalcuSyn software. For in vivo studies, we evaluated the effect of both drugs, alone and in combination, on survival of CCRF-CEM tumor-bearing animals. Mechanistic studies addressed recovery of DNA synthesis, intracellular levels of araC metabolites, feedback inhibition by triphosphate species and pharmacokinetics of both drugs.

Results

The combination of troxacitabine and araC in vitro was synergistic with combination indices between 0.1 and 0.7. This appeared to be related to the impact of the combination on DNA synthesis recovery, which was significantly delayed following exposure to the combination of troxacitabine and araC compared to either agent alone. Analysis of the effect of troxacitabine on the intracellular metabolites of araC revealed that troxacitabine did not inhibit araC deamination and caused a slight decrease in the overall intracellular accumulation of araCTP. The lower accumulation of araCTP could not be attributed to feedback inhibition caused by troxacitabine triphosphate on dCK. Furthermore, our in vivo experiments demonstrated that the combination of araC and troxacitabine was better at slowing down the progression of leukemia in SCID mice than either agent used alone without additive toxicities. Injections of 10 mg/kg troxacitabine i.p. daily for 5 days in combination with araC at 10 mg/kg led to an increase in median survival time of 58 days compared to 49.5 and 53.5 days for araC and troxacitabine, respectively, given as single agents. This represents an increase in life span of 17%, respectively when compared to araC alone. A pharmacokinetic study revealed that troxacitabine did not influence the disposition of araC when coadministered.

Conclusions

Overall, our results show that the antileukemic activity of troxacitabine and araC is complementary when the two nucleoside analogues are combined in vivo. These effects appear to be related to their interaction at the level of DNA repair rather than to pharmacokinetic interactions. These results encourage the use of troxacitabine and araC in combination in patients with acute leukemia.

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Fig. 1.
Fig. 2A–C.
Fig. 3.
Fig. 4A, B.

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Abbreviations

AML:

Acute myelogenous leukemia

araC:

1-β-d-Arabinofuranosylcytosine, cytarabine

araCMP:

Cytarabine-5′-monophosphate

araCTP:

Cytarabine-5′-triphosphate

araU:

1-β-d-Arabinofuranosyluracil

dCyd:

Deoxycytidine

CDA:

Cytidine deaminase

CI:

Combination index

dCK:

Deoxycytidine kinase

dCTP:

Deoxycytidine-5′-triphosphate

dFdC:

Gemcitabine, 2′,2′-difluorodeoxycytidine

troxacitabineTP:

Troxacitabine 5′-triphosphate

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Acknowledgements

The authors would like to thank Lucie Bibeau, Chantal Boudreau, Dominique Custeau and Marie-Josée Gilbert for their help with the animal studies. Also, we would like to thank Karen Meerovitch for helpful discussions and revision, and Johanne Cadieux for assistance in the preparation of the manuscript.

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  1. Shire BioChem Inc., 275 Armand-Frappier Blvd., Laval, Québec, H7V 4A7, Canada

    David Y. Bouffard, Jacques Jolivet, Lorraine Leblond, Bettina Hamelin, France Ouellet, Sylvain Barbeau, Annie Richard & Henriette Gourdeau

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  1. David Y. Bouffard
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  2. Jacques Jolivet
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Correspondence to Henriette Gourdeau.

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Bouffard, D.Y., Jolivet, J., Leblond, L. et al. Complementary antineoplastic activity of the cytosine nucleoside analogues troxacitabine (Troxatyl) and cytarabine in human leukemia cells. Cancer Chemother Pharmacol 52, 497–506 (2003). https://doi.org/10.1007/s00280-003-0699-4

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