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An improved human carboxylesterase for enzyme/prodrug therapy with CPT-11

Cancer Gene Therapy volume 15, pages 183–192 (2008)Cite this article

Abstract

CPT-11 is a potent antitumor agent that is activated by carboxylesterases (CE) and intracellular expression of CEs that can activate the drug results in increased cytotoxicity to the drug. As activation of CPT-11 (irinotecan-7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin) by human CEs is relatively inefficient, we have developed enzyme/prodrug therapy approaches based on the CE/CPT-11 combination using a rabbit liver CE (rCE). However, the in vivo application of this technology may be hampered by the development of an immune response to rCE. Therefore, we have developed a mutant human CE (hCE1m6), based on the human liver CE hCE1, that can activate CPT-11 approximately 70-fold more efficiently than the wild-type protein and can be expressed at high levels in mammalian cells. Indeed, adenoviral-mediated delivery of hCE1m6 with human tumor cells resulted in up to a 670-fold reduction in the IC50 value for CPT-11, as compared to cells transduced with vector control virus. Furthermore, xenograft studies with human tumors expressing hCE1m6 confirm the ability of this enzyme to activate CPT-11 in vivo and induce antitumor activity. We propose that this enzyme should likely be less immunogenic than rCE and would be suitable for the in vivo application of CE/CPT-11 enzyme/prodrug therapy.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

CE:

carboxylesterase

CPT-11:

irinotecan,7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin

hCE1:

human carboxylesterase 1

hCE1m6:

hCE1 containing 8 site specific mutations

hiCE:

human intestinal carboxylesterase

IC50:

the concentration of drug that produced a 50% inhibition of cell growth

rCE:

rabbit liver carboxylesterase

SN-38:

7-ethyl-10-hydroxycamptothecin

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Acknowledgements

We thank Dr J Patrick McGovren for the gift of CPT-11 and Dr Randy M Wadkins for suggesting the idea of loop replacement between the two CE proteins. This work was supported in part by NIH Grants CA76202 (PMP), CA79763 (MKD), CA98468 (MRR), CA108775 (PMP), DA18116 (PMP), CA23099 (PJH), a Cancer Center Core Grant P30 CA 21765 and by the American Lebanese Syrian Associated Charities.

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Authors and Affiliations

  1. Department of Molecular Pharmacology, St Jude Children's Research Hospital, Memphis, TN, USA

    M Wierdl, L Tsurkan, J L Hyatt, C C Edwards, M J Hatfield, C L Morton, P J Houghton, M K Danks & P M Potter

  2. Department of Chemistry and Biochemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    M R Redinbo

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  1. M Wierdl
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Correspondence to P M Potter.

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Wierdl, M., Tsurkan, L., Hyatt, J. et al. An improved human carboxylesterase for enzyme/prodrug therapy with CPT-11. Cancer Gene Ther 15, 183–192 (2008). https://doi.org/10.1038/sj.cgt.7701112

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