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Temozolomide Delivery to Tumor Cells by a Multifunctional Nano Vehicle Based on Poly(β-L-malic acid)

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ABSTRACT

Purpose

Temozolomide (TMZ) is a pro-drug releasing a DNA alkylating agent that is the most effective drug to treat glial tumors when combined with radiation. TMZ is toxic, and therapeutic dosages are limited by severe side effects. Targeted delivery is thus needed to improve efficiency and reduce non-tumor tissue toxicity.

Methods

Multifunctional targetable nanoconjugates of TMZ hydrazide were synthesized using poly(β-L-malic acid) platform, which contained a targeting monoclonal antibody to transferrin receptor (TfR), trileucine (LLL), for pH-dependent endosomal membrane disruption, and PEG for protection.

Results

The water-soluble TMZ nanoconjugates had hydrodynamic diameters in the range of 6.5 to 14.8 nm and ζ potentials in the range of −6.3 to −17.7 mV. Fifty percent degradation in human plasma was observed in 40 h at 37°C. TMZ conjugated with polymer had a half-life of 5–7 h, compared with 1.8 h for free TMZ. The strongest reduction of human brain and breast cancer cell viability was obtained by versions of TMZ nanoconjugates containing LLL and anti-TfR antibody. TMZ-resistant cancer cell lines were sensitive to TMZ nanoconjugate treatment.

Conclusions

TMZ-polymer nanoconjugates entered the tumor cells by receptor-mediated endocytosis, effectively reduced cancer cell viability, and can potentially be used for targeted tumor treatment.

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Abbreviations

TMZ:

temozolomide

TMZH:

temozolomide hydrazide

PMLA:

Poly(β-L-malic acid)

HuTfR:

anti-human transferrin receptor

mAb:

monoclonal antibody

LOEt:

L-leucine ethyl ester

LLL:

L-leucine tripeptide

Alex680:

Alexa Fluor® 680 C2 maleimide

 :

Nanoconjugates are abbreviated as shown in Charts 3 and 4

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ACKNOWLEDGEMENTS

We greatly acknowledge financial support by NIH R01 CA123495 and grant from the Department of Neurosurgery at Cedars-Sinai Medical Center. Dr. Reinhard Sterner, Institut für Biophysik und physikalische Biochemie der Universität Regensburg, Regensburg, Germany supported the production of polymalic acid.

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Authors and Affiliations

  1. Department of Neurosurgery, Cedars-Sinai Medical Center, 8631 W. Third Street, Suite 800E, Los Angeles, California, 90048, USA

    Rameshwar Patil, José Portilla-Arias, Hui Ding, Satoshi Inoue, Bindu Konda, Jinwei Hu, Keith L. Black, Eggehard Holler & Julia Y. Ljubimova

  2. Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California, USA

    Kolja A. Wawrowsky

  3. Department of Chemistry and Biochemistry, California State University, Northridge, California, USA

    Paul K. Shin

  4. Institut für Biophysik und Physikalische Biochemie der Universität Regensburg, Regensburg, Germany

    Eggehard Holler

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  1. Rameshwar Patil
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Correspondence to Julia Y. Ljubimova.

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Patil, R., Portilla-Arias, J., Ding, H. et al. Temozolomide Delivery to Tumor Cells by a Multifunctional Nano Vehicle Based on Poly(β-L-malic acid). Pharm Res 27, 2317–2329 (2010). https://doi.org/10.1007/s11095-010-0091-0

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