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The metabolic profile of mitoxantrone and its relation with mitoxantrone-induced cardiotoxicity

  • Organ Toxicity and Mechanisms
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Abstract

Mitoxantrone (MTX) is an antitumor agent that causes cardiotoxicity in 18 % patients. The metabolic profile of MTX was assessed after incubation of 100 μM MTX with hepatic S9 fraction isolated from rats. The presence of MTX and its metabolites was also assessed in vivo through the analysis of liver and heart extracts of MTX-treated rats. The cytotoxic effects of MTX and MTX metabolites were evaluated in the H9c2 cells after 24-h incubation with MTX alone and MTX + metabolites. The influence of CYP450- and CYP2E1-mediated metabolism for the cytotoxicity of MTX was assessed after 96-h incubation with MTX (100 nM and 1 μM) in the presence/absence of CYP450 or CYP2E1 inhibitors. After 4-h incubation in supplemented S9 fraction, the MTX content was 35 % lower and 5 metabolites were identified: an acetoxy ester derivative (never described before), two glutathione conjugates, a monocarboxylic acid derivative, and the naphtoquinoxaline, the later commonly related to MTX pharmacological effects. The presence of MTX and naphtoquinoxaline metabolite was evidenced in vivo in liver and heart of MTX-treated rats. The cytotoxicity caused by MTX + metabolites was higher than that observed in the H9c2 cells incubated with non-metabolized MTX group. The co-incubation of MTX with CYP450 and CYP2E1 inhibitors partially prevented the cytotoxicity observed in the MTX groups incubated with H9c2 cells, highlighting that the metabolism of MTX is relevant for its undesirable effects. The naphtoquinoxaline metabolite is described in heart and liver in vivo, highlighting that this metabolite accumulates in these tissues. It was demonstrated that MTX P450-mediated metabolism contributed to MTX toxicity.

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Abbreviations

DAD:

Photodiode array

DAS:

Diallyl sulfide

DMEM:

Dulbecco’s modified eagle’s medium

DMSO:

Dimethyl sulfoxide

ESI:

Electrospray ionization interface

GSH:

Reduced glutathione

LC:

Liquid chromatography

MS:

Mass spectrometry

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

MTP:

Metyrapone

MTX:

Mitoxantrone

β-NADPH:

β-Nicotinamide adenine dinucleotide phosphate

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Acknowledgments

Authors are grateful to Dr. Zelia dos Santos Azevedo, from Faculty of Sciences, University of Porto, for gently lend us the LC/DAD-ESI/MS and for all precious technical assistance. This work was supported by the Fundação para a Ciência e Tecnologia (FCT)—project [EXPL/DTP-FTO/0290/2012]—QREN initiative with EU/FEDER financing through COMPETE—Operational Programme for Competitiveness Factors. LGR and VMC thank FCT for their Ph.D. grant (SFRH/BD/63473/2009) and Post-doc grant (SFRH/BPD/63746/2009), respectively. The authors are also grateful to Fundação para a Ciência e a Tecnologia (FCT) for Grant No. PEst-C/EQB/LA0006/2011

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Authors and Affiliations

  1. REQUIMTE, Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Rua Jorge Viterbo Ferreira, 228, Porto, 4050-313, Portugal

    Luciana Grazziotin Rossato, Vera Marisa Costa, Paula Guedes de Pinho, Marcelo Dutra Arbo, Laure Vilain, Maria de Lourdes Bastos & Fernando Remião

  2. Centro de Investigação em Química, Departamento de Química, Faculdade de Ciências, Universidade do Porto, Porto, Portugal

    Victor de Freitas

  3. Centro de Neurociências e Biologia Celular de Coimbra, Departamento de Ciências da Vida, Universidade de Coimbra, Coimbra, Portugal

    Carlos Palmeira

Authors
  1. Luciana Grazziotin Rossato
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  2. Vera Marisa Costa
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  3. Paula Guedes de Pinho
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Correspondence to Luciana Grazziotin Rossato or Fernando Remião.

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Rossato, L.G., Costa, V.M., de Pinho, P.G. et al. The metabolic profile of mitoxantrone and its relation with mitoxantrone-induced cardiotoxicity. Arch Toxicol 87, 1809–1820 (2013). https://doi.org/10.1007/s00204-013-1040-6

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