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Synthesis and Εvaluation of Αnticancer Αctivity in Cells of Novel Stoichiometric Pegylated Fullerene-Doxorubicin Conjugates

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Abstract

Purpose

To synthesize pegylated stoichiometrically and structurally well-defined conjugates of fullerene (C60) with doxorubicin (DOX) and investigate their antiproliferative effect against cancer cell lines.

Methods

Stoichiometric (1:1 and 1:2) pegylated conjugates of C60 with DOX were synthesized using the Prato reaction to create fulleropyrrolidines equipped with a carboxyl function for anchoring a polyethylene glycol (PEG) moiety and either a hydroxyl group for attaching one molecule of DOX or a terminal alkyne group for attaching two molecules of DOX through a click reaction. In both conjugates, the DOX moieties are held through a urethane-type bond. Drug release was studied in phosphate buffer (PBS, pH 7.4) and MCF-7 cancer cells lysate. The uptake of the conjugates by MCF-7 cancer cells and their intracellular localization were studied with fluorescence microscopy. The antiproliferative activity of the conjugates was investigated using the WST-1 test.

Results

One or two DOX molecules were anchored on pegylated C60 particles to form DOX-C60-PEG conjugates. Drug liberation from the conjugates was significantly accelerated in the presence of tumor cell lysate compared to PBS. The conjugates could be internalized by MCF-7 cells. DOX from the conjugates exhibited much delayed, compared to free DOX, localization in the nucleus and antiproliferative activity.

Conclusion

Pegylated DOX-C60 conjugates (1:1) and (2:1) with well-defined structure were successfully synthesized and found to exhibit comparable, but with a delayed onset, antiproliferative activity with free DOX against MCF-7 cancer cells. The results obtained justify further investigation of the potential of these conjugates as anticancer nanomedicines.

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Abbreviations

DIAD:

diisopropyl azodicarboxylate

DIC:

N,N′-diisopropylcarbodiimide

FCC:

Flash Column Chromatography

HBTU:

O-(benzotriazol-1-yl)-N,N,N΄,N΄-tetramethyluronium hexafluorophosphate

HOSu:

N-hydroxysuccinimide

MeO-PEG-NH2 :

amino-polyethylene glycol monomethyl ether

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ACKNOWLEDGMENTS AND DISCLOSURES

We wish to thank Dr. N. Eilert, University of Leipsig, for guidance regarding the preliminary biological results, Dr. G. Tsivgoulis, and Dr. K. Andriopoulou, Department of Chemistry, University of Patras, for UV/vis spectra and TGA measurements and related comments, respectively.

Author information

Authors and Affiliations

  1. Laboratory of Synthetic Organic Chemistry, Department of Chemistry, University of Patras, 26504, Patras, Greece

    George E. Magoulas & Dionissios Papaioannou

  2. Department of Pharmacy, University of Patras, 26504, Patras, Greece

    Marina Bantzi, Danai Messari, Efstathia Voulgari & Konstantinos Avgoustakis

  3. Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504, Patras, Greece

    Chrisostomi Gialeli, Despoina Barbouri & Nikos K. Karamanos

  4. Institut für Organische Chemie, Fakultät für Chemie und Mineralogie, Universität Leipzig, Johannisallee 29, 04103, Leipzig, Germany

    Athanassios Giannis

Authors
  1. George E. Magoulas
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  10. Konstantinos Avgoustakis
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Correspondence to Konstantinos Avgoustakis.

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ESM 1

Extended structures and UV/Vis, TGA, and 1H-NMR data of final products, experimental procedures and characterization for intermediates as well as information on the solubility of compounds 26, 6c and of a non-pegylated fullerene-DOX conjugate are given. Also, the antiproliferative effect of blank (non-loaded with DOX) pegylated C60 particles (compound 26) is presented. (DOC 471 kb)

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Magoulas, G.E., Bantzi, M., Messari, D. et al. Synthesis and Εvaluation of Αnticancer Αctivity in Cells of Novel Stoichiometric Pegylated Fullerene-Doxorubicin Conjugates. Pharm Res 32, 1676–1693 (2015). https://doi.org/10.1007/s11095-014-1566-1

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  • DOI: https://doi.org/10.1007/s11095-014-1566-1

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