• Muluneh Fromsa Seifu Department of Pharmaceutical Sciences, Dibrugarh University
  • L.K. Nath Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh-786004, India
  • Debashis Dutta Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh-786004, India



Objective: Docetaxel, a potent anticancer drug, is suffering from non-specificity and drug resistance as major limitations. In this investigation we developed Hyaluronic acid-Docetaxel conjugate (HA-DTX) loaded nanoliposomes to target cancer cells via passive and active targeting approaches.

Methods: HA-DTX was synthesized and characterized by UV-Visible spectrophotometry, FT-IR spectroscopy, 1H NMR spectroscopy, Differential scanning calorimetry and X-ray diffraction and then loaded into nanoliposomes (L-NLs) by thin film hydration method. L-NLs were characterized physicochemically and evaluated for anticancer efficacy by in vitro cytotoxicity study in glioma cells (C6 glial cells); cellular uptake and apoptotic effect were investigated by fluorescence microscopy.

Results: HA-DTX was successfully synthesized; L-NLs had an average size of 123.0±16.53 nm, polydispersity index of 0.246 and zeta potential of 44.4±6.79 mV. Also, L-NLs exhibited 90.54%±4.22 of drug loading efficiency and 2.68%±0.12 of drug loading releasing about 57.72%±1.17 at pH 5.2 and only 14.14%±1.32 at pH 7.4 after 48 h. No significant change in stability was observed after storage at 5 oC ± 3 oC as well as at 25 °C ± 2 °C/60% RH ± 5% RH for six months. The cytotoxicity effect of L-NLs was 10% higher than that of marketed formulation (percent cell viability: 15.69%±0.72 and 26.50%±0.35, respectively) at 10 µg/ml docetaxel concentration. Fluorescence microscopic investigation showed more cellular uptake and apoptotic effect were observed in L-NLs treated C6 glial cells than those treated with marketed formulation.

Conclusion: HA-DTX loaded nanoliposomes enabled docetaxel to target C6 glial cells with better efficacy and might be effective to treat glioma.

Keywords: Anticancer drug, Hyaluronic acid-Docetaxel conjugate, nanoliposomes, passive and active targeting, C6 glial cells

Keywords: Anticancer drug, Hyaluronic acid-Docetaxel conjugate, nanoliposomes, passive and active targeting, C6 glial cells


1. WHO (World Health Organization). Cancer. [Accessed on 2019 Dec 14]: https://www.who.int/health-topics/cancer#tab=tab_1
2. Ying M, Zhan C, Wang S, Yao B, Hu X, Song X, et al. Liposome-Based Systemic Glioma-Targeted Drug Delivery Enabled by All-D Peptides. ACS Appl Mater Interfaces 2016; 8(44): 29977?29985.
3. Vieira DB, Gamarra LF. Getting into the brain: liposome-based strategies for effective drug delivery across the blood–brain barrier. Int J Nanomedicine 2016; 11: 5381–5414.
4. Persaud-Sharma D, Burns J, Trangle J, Moulik S. Disparities in Brain Cancer in the United States: A Literature Review of Gliomas. Med Sci (Basel) 2017; 5(3): 16.
5. Bastien JIL, McNeill KA, Fine HA. Molecular characterizations of glioblastoma, targeted therapy, and clinical results to date. Cancer 2015; 121(4): 502–516.
6. Djedid R, Kiss R, Lefranc F. Targeted therapy of glioblastomas: a 5-year view. Ther 2009; 6(3): 351–370.
7. Hayward S, Wilson CL, Kidamb S. Hyaluronic acid-conjugated liposome nanoparticles for targeted delivery to CD44 overexpressing glioblastoma cells. Oncotarget 2016; 7 (23): 341–358.
8. Satapathy BS, Mukherjee B, Baishya R, Debnath MC, Dey NS, Maji R. Lipid nanocarrier-based transport of docetaxel across blood brain barrier. RSC Adv 2016; 6: 85261-85274.
9. Kobayashi H, Watanabe R, Choyke PL. Improving conventional enhanced permeability and retention (EPR) effects; what is the appropriate target?. Theranostics 2014; 4: 81–89.
10. Masserini M. Nanoparticles for Brain Drug Delivery. ISRN Biochem 2013; 2013: 1–18.
11. Lam YMF, Chan CYJ, Kuhn JG. Pharmacokinetics and pharmacodynamics of the taxanes. J Oncol Pharm Pract 1997; 2: 78-93.
12. Naguib YW, Rodriguez BL, Li X, Li X, Hursting SD, Williams RO, et al. Solid Lipid Nanoparticle Formulations of Docetaxel Prepared with High Melting Point Triglycerides: In Vitro and in Vivo Evaluation. Mol Pharm 2014; 11: 1239?1249.
13. Kenmotsu H, Tanigawara Y. Pharmacokinetics, dynamics and toxicity of docetaxel: Why the Japanese dose differs from the Western dose. Cancer Sci 2015; 106: 497–504.
14. Pooja D, Kulhari H, Adams DJ, Sistla R. Formulation and dosage of therapeutic nanosuspension for active targeting of docetaxel (WO 2014210485A1). Expert Opin Ther Targets 2016; 26(7): 745-749.
15. Clarke SJ, Rivory LP. Clinical Pharmacokinetics of Docetaxel. Clin Pharmacokinet 1999; 36 (2): 99-114.
16. Sanchez-Moreno P, Boulaiz H, Ortega-Vinuesa JL. Novel drug delivery system based on Docetaxel-loaded nanocapsules as a therapeutic strategy against breast cancer cells. Int J Mol Sci 2012; 13: 4906-4919.
17. Rajappa S, Joshi A, Dova D, Batra U, Rajendranath R, Deo A, et al. Novel formulations of docetaxel, paclitaxel and doxorubicin in the management of metastatic breast cancer. Oncol Lett 2018; 16(3): 3757-3769.
18. Wadhwa S, Mumper RJ. Polymer-drug conjugates for anticancer drug delivery. Crit Rev Ther Drug Carrier Syst 2015; 32(3): 215–245.
19. Haag R, Kratz F. Polymer therapeutics: concepts and applications. Angew Chem Int Ed Engl 2006; 45(8): 1198–1215.
20. Seifu MF, Nath LK. Polymer-drug conjugates: novel carriers for cancer chemotherapy. Polym Plast Technol Eng 2019; 58(2): 158-171.
21. Luo Y, Ziebell MR, Prestwich GD. A Hyaluronic acid-taxol antitumor bioconjugate targeted to cancer cells. Biomacromolecules 2000; 1: 208-218.
22. Tripodo G, Trapani A, Torre ML, Giammona G, Trapani G, Mandracchia D. Hyaluronic acid and its derivatives in drug delivery and imaging: recent advances and challenges. Eur J Pharm Biopharm 2015; 97: 400-416.
23. Luo Y, Prestwich GD. Hyaluronic acid-N-hydroxysuccinimide: A useful intermediate for bioconjugation. Bioconjugate Chem 2001; 12: 1085–1088.
24. Choia KY, Saravanakumar G, Park JH, Park K. Hyaluronic acid-based nanocarriers for intracellular targeting: Interfacial interactions with proteins in cancer. Colloids Surf B 2012; 99: 82– 94.
25. Zhang H, Li Ry, Lu X, Mou Z, Lin G. Docetaxel-loaded liposomes. Preparation, pH sensitivity, pharmacokinetics, and tissue distribution. J Zhejiang Univ Sci B 2012; 13(12): 981-989.
26. Shaw TK, Mandal D, Dey G, Pal MM, Paul P, Chakraborty S, et al. Successful delivery of docetaxel to rat brain using experimentally developed nanoliposome: a treatment strategy for brain tumor. Drug Deliv 2017; 24(1): 346-357.
27. Mittapalli RK, Liu X, Adkins CE, Nounou MI, Bohn KA, Terrell TB, et al. Paclitaxel–Hyaluronic NanoConjugates Prolong Overall Survival in Preclinical Brain Metastases of Breast Cancer Model. Mol Cancer Ther 2013; 12(11): OF1-OF11.
28. Dubey RD, Klippstein R, Wang JT, Hodgins N, Mei K, Sosabowski J, et al. Novel hyaluronic acid conjugates for dual nuclear imaging and therapy in CD44-expressing tumors in mice in vivo. Nanotheranostics 2017; 1(1): 59–79.
29. Dey NS, Mukherjee B, Maji R, Satapathy BS. Development of linker conjugated nanosize lipid vesicles: a strategy for cell selective treatment in breast cancer. Curr Cancer Drug Targets 2016; 16: 357–372.
30. Ren G, Liu D, Guo W, Wang M, Wu C, Guo M, et al. Docetaxel prodrug liposomes for tumor therapy: characterization, in vitro and in vivo evaluation. Drug Deliv 2016; 23(4): 1272–1281.
31. European Medicines Agency (EMEA). Stability testing of new drug substances and products (ICH Topic Q1A (R2)). London: 2006, p. 14.
32. Barth RF. Rat brain tumor models in experimental neurooncology: the 9L, C6, T9, F98, RG2 (D74), RT-2 and CNS-1 gliomas. J Neuro-oncol 1998; 36: 91–102.
33. Thermo Fisher Scientific (Invitrogen). User Guide for CellEventTM Caspase-3/7 Green Detection Reagent. Carlsbad (CA): Thermo Fisher Scientific Inc; 2017. p. 1-2. Catalog No. C10423, C10723, Pub. No. MAN0003556, Rev. B.0,
34. Wolny PM, Banerji S, Gounou C, Brisson AR, Day AJ, Jackson DG, et al. Analysis of CD44-hyaluronan interactions in an artificial membrane system: insights into the distinct binding properties of high and low molecular weight hyaluronan. J Biol Chem 2010; 285: 30170-30180.
35. Fan X, Zhao X, Qu X, Fang J. pH sensitive polymeric complex of cisplatin with hyaluronic acid exhibits tumor-targeted delivery and improved in vivo antitumor effect. Int J Pharm 2015; 496: 644-653.
36. Leonelli F, Bella AL, Migneco LM, Bettolo RM. Design, Synthesis and Applications of Hyaluronic Acid-Paclitaxel Bioconjugates. Molecules 2008; 13: 360-378.
37. Lee H, Lee K, Park TG. Hyaluronic acid-Paclitaxel conjugate micelles: synthesis, characterization, and antitumor activity. Bioconjugate Chem 2008; 19: 1319–1325.
38. Goodarzi N, Ghahremani MH, Amini M, et al. CD44-targeted docetaxel conjugate for cancer cells and cancer stem-like cells: A novel hyaluronic acid-based drug delivery system. Chem Biol Drug Des 2014; 83: 741–752.
39. Veereshappa, Purohit P, Shrawat VK, Singh VK, Inventors; Shilpa Medicare Limited, assignee. Process for preparing docetaxel trihydrate polymorph. Patent cooperation treaty (PCT), World Intellectual Property Organization, International Bureau WO2012160568A1. 29 November 2012.
40. Tatini LK, Reddy KVSRK, Rao NS. Vapor-Induced Phase Transformations in Docetaxel. AAPS PharmSciTech 2012; 13 (2): 548–555.
41. Bozzuto G, Molinari A. Liposomes as nanomedical devices. Int J Nanomed 2015; 10: 975–999.
42. MacLachlan I. Liposomal formulations for nucleic acid delivery. In: Crooke ST, editor. Antisense drug technology: principles: strategies, and applications. 2nd ed. London: Taylor & Francis Group, LLC; 2007. p. 253–254.
43. Eloya JO, Petrillia R, Topana JF, Antonio HMR, Barcellos JPA, Chesca DL, et al. Co-loaded paclitaxel/rapamycin liposomes: Development, characterization and in vitro and in vivo evaluation for breast cancer therapy. Colloids Surf B 2016; 141: 74–82.
44. Martins KF, Messias AD, Leite FL, Duek EAR Preparation and characterization of paclitaxel-loaded PLDLA microspheres. Mater Res 2014; 17: 650–656.
45. Sibenaller ZA, Etame AB, Ali MM, Barua M, Braun TA, Casavant T L, et al. Genetic characterization of commonly used glioma cell lines in the rat animal model system. Neurosurg Focus 2005; 19(4): 1-9.
46. Beaulieu E, Demeule M, Ghitescu L, Béliveau R. P-glycoprotein is strongly expressed in the luminal membranes of the endothelium of blood vessels in the brain. Biochem J 1997; 326: 539–544.
47. Gottesman MM, Pastan IH. The Role of Multidrug Resistance Efflux Pumps in Cancer: Revisiting a JNCI Publication Exploring Expression of the MDR1 (P-glycoprotein) Gene. J Natl Cancer Inst 2015; 107(9): 1-3.
48. Tije AJ, Verweij JT, Loos WJ, Sparreboom A. Pharmacological effects of formulation vehicles: implications for cancer chemotherapy. Clin Pharmacokinet 2003; 42: 665–685.
49. Mattheolabakis G, Milane L, Singh A, Amiji MM. Hyaluronic acid targeting of CD44 for cancer therapy: from receptor biology to nanomedicine. J Drug Target 2015; 23(7-8): 605-618.
15 Views | Downloads
How to Cite
Fromsa Seifu, M., Lila Kanta, N., & Dutta, D. (2020). HYALURONIC ACID-DOCETAXEL CONJUGATE LOADED NANOLIPOSOMES FOR TARGETING TUMOR CELLS. International Journal of Applied Pharmaceutics, 12(6). Retrieved from https://innovareacademics.in/journals/index.php/ijap/article/view/39026
Original Article(s)