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Pharmaceutical Research

, Volume 32, Issue 5, pp 1676–1693 | Cite as

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

  • George E. Magoulas
  • Marina Bantzi
  • Danai Messari
  • Efstathia Voulgari
  • Chrisostomi Gialeli
  • Despoina Barbouri
  • Athanassios Giannis
  • Nikos K. Karamanos
  • Dionissios Papaioannou
  • Konstantinos AvgoustakisEmail author
Research Paper

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.

KEY WORDS

antiproliferative activity conjugates doxorubicin fullerenes nanoparticles 

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

Notes

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.

Supplementary material

11095_2014_1566_MOESM1_ESM.doc (472 kb)
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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • George E. Magoulas
    • 1
  • Marina Bantzi
    • 2
  • Danai Messari
    • 2
  • Efstathia Voulgari
    • 2
  • Chrisostomi Gialeli
    • 3
  • Despoina Barbouri
    • 3
  • Athanassios Giannis
    • 4
  • Nikos K. Karamanos
    • 3
  • Dionissios Papaioannou
    • 1
  • Konstantinos Avgoustakis
    • 2
    Email author
  1. 1.Laboratory of Synthetic Organic Chemistry, Department of ChemistryUniversity of PatrasPatrasGreece
  2. 2.Department of PharmacyUniversity of PatrasPatrasGreece
  3. 3.Laboratory of Biochemistry, Department of ChemistryUniversity of PatrasPatrasGreece
  4. 4.Institut für Organische Chemie, Fakultät für Chemie und MineralogieUniversität LeipzigLeipzigGermany

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