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Methotrexate binds to recombinant thiopurine S-methyltransferase and inhibits enzyme activity after high-dose infusions in childhood leukaemia

  • Pharmacogenetics
  • Published:
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Abstract

Purpose

Important drugs in the treatment of childhood acute lymphoblastic leukaemia (ALL) are 6-mercaptopurine (6-MP) and methotrexate (MTX). Thiopurine methyltransferase (TPMT) is a polymorphic enzyme causing variability in 6-MP response and toxicity. The aim of this study was to investigate the fluctuation in TPMT enzyme activity over time and the effect of high-dose MTX infusions on TPMT enzyme activity and 6-MP metabolites in paediatric ALL patients.

Methods

Fifty-three children with ALL treated according to the NOPHO-ALL 2000 protocol were included in the study. TPMT enzyme activity was measured at six different times starting from diagnosis until after the end of maintenance treatment. TPMT and 6-MP metabolites were measured before the initiation of high-dose MTX (HD-MTX) infusions and at 66 h post-infusion. The interaction between MTX and TPMT was investigated in vitro using recombinant TPMT protein and a leukaemic cell line.

Results

Forty percent of TPMT wild-type individuals had deceptively low TPMT enzyme activity according to genotype at the time of diagnosis. TPMT activity had decreased significantly 66 h after the start of HD-MTX infusions (−9.2 %; p = 0.013). MTX bound to recombinant TPMT protein severely inhibiting TPMT enzyme activity (remaining activity 16 %).

Conclusions

Our results show that TPMT genotyping should be performed in children with ALL, since 40 % of the children in our study who carried the wild-type TPMT gene were at risk of initial underdosing of 6-MP in cases where only TPMT enzyme activity was determined. MTX inhibits the TPMT enzyme activity after HD-MTX infusions due to protein binding.

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Acknowledgements

This study was supported by The Swedish Childhood Cancer Foundation, The Swedish Cancer Society, The Swedish Medical Research Council and The LiU Cancer Network. We would like to thank the children and their parents/guardians for their participation in the present study as well as the medical and nursing staff who made this work possible. The skilful technical assistance of Karin Skoglund, Britt Sigfridsson and Monica Häger has also been greatly appreciated.

Conflict of interests

None.

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Authors and Affiliations

  1. Department of Physics, Chemistry and Biology, Linköping University, Linköping, Sweden

    Patricia Wennerstrand & Lars-Göran Mårtensson

  2. Childhood Cancer Research Unit, Department of Women and Child Health, Astrid Lindgren Children’s Hospital, Karolinska University Hospital, Stockholm, Sweden

    Stefan Söderhäll

  3. Division of Drug Research, Department of Medical and Health Sciences, Linköping University, Linköping, Sweden

    Anna Zimdahl & Malin Lindqvist Appell

Authors
  1. Patricia Wennerstrand
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  2. Lars-Göran Mårtensson
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  3. Stefan Söderhäll
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  4. Anna Zimdahl
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  5. Malin Lindqvist Appell
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Corresponding author

Correspondence to Malin Lindqvist Appell.

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Wennerstrand, P., Mårtensson, LG., Söderhäll, S. et al. Methotrexate binds to recombinant thiopurine S-methyltransferase and inhibits enzyme activity after high-dose infusions in childhood leukaemia. Eur J Clin Pharmacol 69, 1641–1649 (2013). https://doi.org/10.1007/s00228-013-1521-9

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  • DOI: https://doi.org/10.1007/s00228-013-1521-9

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