Transferrin-Modified Vitamin-E/Lipid Based Polymeric Micelles for Improved Tumor Targeting and Anticancer Effect of Curcumin

  • Omkara Swami Muddineti
  • Preeti Kumari
  • Balaram Ghosh
  • Swati Biswas
Research Paper



Transferrin receptor (TfR) is up-regulated in various malignant tumors not only to meet the iron requirement, but also to increase the cell survival via participation in various cellular signaling pathways. Here we explored transferrin as ligand for Poly(ethylene Glycol) (PEG)-ylated vitamin-E/lipid (PE) core micelles (VPM).


Transferrin modified polymer was synthesized and drug loaded micelles were evaluated in 2D Hela and HepG2 cancer cells for cellular uptake and cytotoxicity and in 3D Hela spheroids for growth inhibition, uptake and penetration studies.


Targeted (Tf-VPM) and non-targeted (VPM) micelles showed mean hydrodynamic diameter of 114.2 ± 0.64 nm and 117.4 ± 0.72 nm and zeta potential was −22.8 ± 0.62 and −14.8 ± 1.74 mV, respectively. Cellular uptake study indicated that the Tf-CVPM were taken up by cancer cells (Hela and HepG2) with higher efficiency. Enhanced cytotoxicity was demonstrated for Tf-VPM compared to CVPM. Marked spheroid growth inhibition following treatment with Tf-CVPM was observed compared to the treatment with non-targeted CVPM.


The developed transferrin-modified micelles have improved ability to solubilize the loaded drugs and could actively target solid tumors by its interaction with over-expressed transferrin receptors. Therefore, the nano-micelles could be further explored for its potential utilization in cancer therapy.

Key words

active targeting curcumin micelles transferrin vitamin E 





Curcumin loaded VPM


Dulbecco’s modified Eagle’s media


Dioleoyl phosphatidylethanolamine


Heat-inactivated fetal bovine serum


Free curcumin




Human cervical carcinoma cells


Human hepatic carcinoma cells


Minimuim essential medium


Dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide


Polyethylene glycol-phosphatidyl ethanolamine




PEG-PE based micelles




Curcumin loaded Tf-VPM


Transferrin modified PEG-PE


Transferrin modified Vitamin E based polymer


Transferrin modified vitamin E based micelles


Vitamin E based micelles



The work was supported in part by the grants provided by the Department of Science and Technology (CS-269/2013), Government of India and the Department of Biotechnology (BT/Bio-CARe/07/10003/2013–14), Govt of India to Swati Biswas. Omkara Swami gratefully acknowledges Indian Council of Medical Research (2014–24,190), Department of Health Research, Ministry of Health & Family Welfare, Government of India for awarding him with the Senior Research Fellowship (SRF). There are no conflicts of interest

Supplementary material

11095_2018_2382_MOESM1_ESM.docx (321 kb)
ESM 1 (DOCX 320 kb)


  1. 1.
    Muddineti OS, Ghosh B, Biswas S. Current trends in using polymer coated gold nanoparticles for cancer therapy. Int J Pharm. 2015;484(1):252–67.CrossRefPubMedGoogle Scholar
  2. 2.
    Xin Y, Huang Q, Tang J-Q, Hou X-Y, Zhang P, Zhang LZ, et al. Nanoscale drug delivery for targeted chemotherapy. Cancer Lett. 2016;379(1):24–31.CrossRefPubMedGoogle Scholar
  3. 3.
    Schmitz N, Pfistner B, Sextro M, Sieber M, Carella AM, Haenel M, et al. Aggressive conventional chemotherapy compared with high-dose chemotherapy with autologous haemopoietic stem-cell transplantation for relapsed chemosensitive Hodgkin's disease: a randomised trial. Lancet. 2002;359(9323):2065–71.CrossRefPubMedGoogle Scholar
  4. 4.
    Shi C, Zhang Z, Shi J, Wang F, Luan Y. Co-delivery of docetaxel and chloroquine via PEO–PPO–PCL/TPGS micelles for overcoming multidrug resistance. Int J Pharm. 2015;495(2):932–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Abdullin TI, Bondar OV, Nikitina II, Bulatov ER, Morozov MV, Hilmutdinov A, et al. Effect of size and protein environment on electrochemical properties of gold nanoparticles on carbon electrodes. Bioelectrochemistry. 2009;77(1):37–42.CrossRefPubMedGoogle Scholar
  6. 6.
    Ford JM, Hait WN. Pharmacology of drugs that alter multidrug resistance in cancer. Pharmacol Rev. 1990;42(3):155–99.PubMedGoogle Scholar
  7. 7.
    te Velde EA, Vogten JM, Gebbink MF, van Gorp JM, Voest EE, Borel Rinkes I. Enhanced antitumour efficacy by combining conventional chemotherapy with angiostatin or endostatin in a liver metastasis model. Br J Surg. 2002;89(10):1302–9.CrossRefGoogle Scholar
  8. 8.
    Maeda H, Wu J, Sawa T, Matsumura Y, Hori K. Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J Control Release. 2000;65(1):271–84.CrossRefPubMedGoogle Scholar
  9. 9.
    Maeda H, Bharate G, Daruwalla J. Polymeric drugs for efficient tumor-targeted drug delivery based on EPR-effect. Eur J Pharm Biopharm. 2009;71(3):409–19.CrossRefPubMedGoogle Scholar
  10. 10.
    Atta AH, El-Shenawy AI, Refat MS, Elsabawy KM. Preparation and characterization of some gold nanometric compounds with simple organic materials as precursor: spectroscopic, biological and anti-cancer assessments. J Mol Struct. 2013;1039:51–60.CrossRefGoogle Scholar
  11. 11.
    Biswas S, Torchilin VP. Nanopreparations for organelle-specific delivery in cancer. Adv Drug Deliv Rev. 2014;66:26–41.CrossRefPubMedGoogle Scholar
  12. 12.
    Chandrasekharan P, Maity D, Yong CX, Chuang K-H, Ding J, Feng S-S. Vitamin E (d-alpha-tocopheryl-co-poly(ethylene glycol) 1000 succinate) micelles-superparamagnetic iron oxide nanoparticles for enhanced thermotherapy and MRI. Biomaterials. 2011;32(24):5663–72.CrossRefPubMedGoogle Scholar
  13. 13.
    Kutty RV, Feng S-S. Cetuximab conjugated vitamin E TPGS micelles for targeted delivery of docetaxel for treatment of triple negative breast cancers. Biomaterials. 2013;34(38):10160–71.CrossRefPubMedGoogle Scholar
  14. 14.
    Muddineti OS, Ghosh B, Biswas S. Current trends in the use of vitamin E-based micellar nanocarriers for anticancer drug delivery. Expert Opin Drug Deliv. 2016;14(6):1–12.Google Scholar
  15. 15.
    Li P-Y, Lai P-S, Hung W-C, Syu W-J. Poly (L-lactide)-vitamin E TPGS nanoparticles enhanced the cytotoxicity of doxorubicin in drug-resistant MCF-7 breast cancer cells. Biomacromolecules. 2010;11(10):2576–82.CrossRefPubMedGoogle Scholar
  16. 16.
    Hayashi T, Tsai S-Y, Mori T, Fujimoto M, Su T-P. Targeting ligand-operated chaperone sigma-1 receptors in the treatment of neuropsychiatric disorders. Expert Opin Ther Targets. 2011;15(5):557–77.CrossRefPubMedGoogle Scholar
  17. 17.
    Sriraman SK, Pan J, Sarisozen C, Luther E, Torchilin V. Enhanced cytotoxicity of folic acid-targeted liposomes co-loaded with C6 ceramide and doxorubicin: in vitro evaluation on HeLa, A2780-ADR, and H69-AR cells. Mol Pharm. 2016;13(2):428–37.CrossRefPubMedGoogle Scholar
  18. 18.
    Zeng X, Sun Y-X, Qu W, Zhang X-Z, Zhuo R-X. Biotinylated transferrin/avidin/biotinylated disulfide containing PEI bioconjugates mediated p53 gene delivery system for tumor targeted transfection. Biomaterials. 2010;31(17):4771–80.CrossRefPubMedGoogle Scholar
  19. 19.
    Tortorella S, Karagiannis TC. Transferrin receptor-mediated endocytosis: a useful target for cancer therapy. J Membr Biol. 2014;247(4):291–307.CrossRefPubMedGoogle Scholar
  20. 20.
    Yue J, Liu S, Wang R, Hu X, Xie Z, Huang Y, et al. Transferrin-conjugated micelles: enhanced accumulation and antitumor effect for transferrin-receptor-overexpressing cancer models. Mol Pharm. 2012;9(7):1919–31.CrossRefPubMedGoogle Scholar
  21. 21.
    Abouzeid AH, Patel NR, Sarisozen C, Torchilin VP. Transferrin-targeted polymeric micelles co-loaded with curcumin and paclitaxel: efficient killing of paclitaxel-resistant cancer cells. Pharm Res. 2014;31(8):1938–45.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Chen H, Zhang T, Zhou Z, Guan M, Wang J, Liu L, et al. Enhanced uptake and cytotoxity of folate-conjugated mitoxantrone-loaded micelles via receptor up-regulation by dexamethasone. Int J Pharm. 2013;448(1):142–9.CrossRefPubMedGoogle Scholar
  23. 23.
    Nam J-P, Park S-C, Kim T-H, Jang J-Y, Choi C, Jang M-K, et al. Encapsulation of paclitaxel into lauric acid-O-carboxymethyl chitosan-transferrin micelles for hydrophobic drug delivery and site-specific targeted delivery. Int J Pharm. 2013;457(1):124–35.CrossRefPubMedGoogle Scholar
  24. 24.
    Zhang P, Hu L, Yin Q, Zhang Z, Feng L, Li Y. Transferrin-conjugated polyphosphoester hybrid micelle loading paclitaxel for brain-targeting delivery: synthesis, preparation and in vivo evaluation. J Control Release. 2012;159(3):429–34.CrossRefPubMedGoogle Scholar
  25. 25.
    Muthu MS, Kutty RV, Luo Z, Xie J, Feng S-S. Theranostic vitamin E TPGS micelles of transferrin conjugation for targeted co-delivery of docetaxel and ultra bright gold nanoclusters. Biomaterials. 2015;39:234–48.CrossRefPubMedGoogle Scholar
  26. 26.
    Muddineti OS, Kumari P, Ghosh B, Torchilin VP, Biswas S. d-α-Tocopheryl succinate/Phosphatidyl ethanolamine conjugated amphiphilic polymer-based Nanomicellar system for the efficient delivery of curcumin and to overcome multiple drug resistance in cancer. ACS Appl Mater Interfaces. 2017;9:16778–92.CrossRefPubMedGoogle Scholar
  27. 27.
    Torchilin V. Targeted polymeric micelles for delivery of poorly soluble drugs. Cell Mol Life Sci. 2004;61(19):2549–59.CrossRefPubMedGoogle Scholar
  28. 28.
    Sarisozen C, Abouzeid AH, Torchilin VP. The effect of co-delivery of paclitaxel and curcumin by transferrin-targeted PEG-PE-based mixed micelles on resistant ovarian cancer in 3-D spheroids and in vivo tumors. Eur J Pharm Biopharm. 2014;88(2):539–50.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Wu H, Zhu L, Torchilin VP. pH-sensitive poly (histidine)-PEG/DSPE-PEG co-polymer micelles for cytosolic drug delivery. Biomaterials. 2013;34(4):1213–22.CrossRefPubMedGoogle Scholar
  30. 30.
    Yang C, Chen H, Zhao J, Pang X, Xi Y, Zhai G. Development of a folate-modified curcumin loaded micelle delivery system for cancer targeting. Colloids Surf B: Biointerfaces. 2014;121:206–13.CrossRefPubMedGoogle Scholar
  31. 31.
    Liang N, Sun S, Li X, Piao H, Piao H, Cui F, et al. Alpha-tocopherol succinate-modified chitosan as a micellar delivery system for paclitaxel: preparation, characterization and in vitro/in vivo evaluations. Int J Pharm. 2012;423(2):480–8.CrossRefPubMedGoogle Scholar
  32. 32.
    Sriraman SK, Salzano G, Sarisozen C, Torchilin V. Anti-cancer activity of doxorubicin-loaded liposomes co-modified with transferrin and folic acid. Eur J Pharm Biopharm. 2016;105:40–9.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Pereira PM, Berisha N, Bhupathiraju NDK, Fernandes R, Tomé JP, Drain CM. Cancer cell spheroids are a better screen for the photodynamic efficiency of glycosylated photosensitizers. PLoS One. 2017;12(5):e0177737.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Li L, Yang Q, Zhou Z, Zhong J, Huang Y. Doxorubicin-loaded, charge reversible, folate modified HPMA copolymer conjugates for active cancer cell targeting. Biomaterials. 2014;35(19):5171–87.CrossRefPubMedGoogle Scholar
  35. 35.
    Liu Z, Gao X, Kang T, Jiang M, Miao D, Gu G, et al. B6 peptide-modified PEG-PLA nanoparticles for enhanced brain delivery of neuroprotective peptide. Bioconjug Chem. 2013;24(6):997–1007.CrossRefPubMedGoogle Scholar
  36. 36.
    Harding C, Heuser J, Stahl P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J Cell Biol. 1983;97(2):329–39.CrossRefPubMedGoogle Scholar
  37. 37.
    McMahon HT, Boucrot E. Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol. 2011;12(8):517–33.CrossRefPubMedGoogle Scholar
  38. 38.
    Danhier F, Kouhé TTB, Duhem N, Ucakar B, Staub A, Draoui N, et al. Vitamin E-based micelles enhance the anticancer activity of doxorubicin. Int J Pharm. 2014;476(1):9–15.CrossRefPubMedGoogle Scholar
  39. 39.
    Tima S, Ampasavate SCA, Berkland C, Okonogi S. Stable curcumin-loaded polymeric micellar formulation for enhancing cellular uptake and cytotoxicity to FLT3 overexpressing EoL-1 leukemic cells. Eur J Pharm Biopharm. 2017;114:57–68.CrossRefPubMedGoogle Scholar
  40. 40.
    Zanoni M, Piccinini F, Arienti C, Zamagni A, Santi S, Polico R, et al. 3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained. Sci Rep. 2016;6:19103.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Sambale F, Lavrentieva A, Stahl F, Blume C, Stiesch M, Kasper C, et al. Three dimensional spheroid cell culture for nanoparticle safety testing. J Biotechnol. 2015;205:120–9.CrossRefPubMedGoogle Scholar
  42. 42.
    Perche F, Torchilin VP. Cancer cell spheroids as a model to evaluate chemotherapy protocols. Cancer Biol Ther. 2012;13(12):1205–13.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Omkara Swami Muddineti
    • 1
  • Preeti Kumari
    • 1
  • Balaram Ghosh
    • 1
  • Swati Biswas
    • 1
  1. 1.Department of PharmacyBirla Institute of Technology & Science-Pilani, Hyderabad CampusHyderabadIndia

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