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Enhancement of intratumoral cyclophosphamide pharmacokinetics and antitumor activity in a P450 2B11-based cancer gene therapy model

Cancer Gene Therapy volume 14, pages 935–944 (2007)Cite this article

Abstract

The therapeutic utility of cytochrome P450-based enzyme prodrug therapy is well established by preclinical studies and in initial clinical trials. The underlying premise of this gene therapy is that intratumoral P450 expression leads to in situ activation of anticancer P450 prodrugs, such as cyclophosphamide (CPA), with intratumoral accumulation of its activated 4-OH metabolite. In mice bearing 9L gliosarcomas expressing the CPA 4-hydroxylase P450 2B6, enhanced tumor apoptosis was observed 48 h after CPA treatment; however, intratumoral 4-OH-CPA levels were indistinguishable from those of P450-deficient tumors, indicating that the bulk of activated CPA is derived from hepatic metabolism. In contrast, in 9L tumors expressing P450 2B11, a low Km CPA 4-hydroxylase, intratumoral 4-OH-CPA levels were higher than in blood, liver and P450-deficient tumors. Intratumoral 4-OH-CPA increased dose-dependently, without saturation at 140 mg kg−1 CPA, suggesting restricted tumor cell permeation of the parent drug. To circumvent this problem, CPA was administered by direct intratumoral injection, which increased the maximum concentration and area under the curve of drug concentration × time (AUC) of intratumoral 4-OH-CPA by 1.8- and 2.7-fold, respectively. An overall 3.9-fold increase in intratumoral 4-OH-CPA AUC, and in antitumor activity, was obtained when CPA release to systemic circulation was delayed using the slow-release polymer poloxamer 407 as vehicle for intratumoral CPA delivery. These findings highlight the advantage of gene therapy strategies that combine low Km P450 prodrug activation enzymes with slow, localized release of P450 prodrug substrates.

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Abbreviations

AUC:

area under the curve of drug concentration × time

BW:

body weight

C max :

maximum concentration

CPA:

cyclophosphamide

DMEM:

Delbecco's modified Eagle's medium

F127:

Pluronic F127 or poloxamer 407 block copolymer

FBS:

fetal bovine serum

IFA:

ifosfamide

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Acknowledgements

DJW is supported by NIH Grant CA49248.

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Author notes

  1. C-S Chen and Y Jounaidi: These authors have contributed equally to this work.

Authors and Affiliations

  1. Division of Cell and Molecular Biology, Department of Biology, Boston University, Boston, MA, USA

    C-S Chen, Y Jounaidi, T Su & D J Waxman

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  1. C-S Chen
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Correspondence to D J Waxman.

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Chen, CS., Jounaidi, Y., Su, T. et al. Enhancement of intratumoral cyclophosphamide pharmacokinetics and antitumor activity in a P450 2B11-based cancer gene therapy model. Cancer Gene Ther 14, 935–944 (2007). https://doi.org/10.1038/sj.cgt.7701092

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