AAPS PharmSciTech

, Volume 19, Issue 3, pp 1084–1092 | Cite as

Fullerenol-Based Intracellular Delivery of Methotrexate: A Water-Soluble Nanoconjugate for Enhanced Cytotoxicity and Improved Pharmacokinetics

  • Shradha Bahuguna
  • Manish Kumar
  • Gajanand Sharma
  • Rajendra Kumar
  • Bhupinder Singh
  • Kaisar Raza
Research Article


Derivatization of fullerenes to polyhydroxylated fullerenes, i.e., fullerenols (FLU), dramatically decreases their toxicity and has been reported to enhance the solubility as well as cellular permeability. In this paper, we report synthesis of FLU as nanocarrier and subsequent chemical conjugation of Methotrexate (MTX) to FLU with a serum-stable and intracellularly hydrolysable ester bond between FLU and MTX. The conjugate was characterized for physiochemical attributes, micromeritics, drug-loading, and drug-release and evaluated for cancer cell-toxicity, cellular-uptake, hemocompatibility, protein binding, and pharmacokinetics. The developed hemocompatible FL-MTX offered lower protein binding vis-à-vis naïve drug and substantially higher drug loading. The conjugate offered pH-dependent release of 38.20 ± 1.19% at systemic pH and 85.67 ± 3.39% at the cancer cell pH. FLU-MTX-treated cells showed significant reduction in IC50 value vis-à-vis the cells treated with pure MTX. Analogously, the results from confocal scanning laser microscopy also confirmed the easy access of the dye-tagged FLU-MTX conjugate to the cell interiors. In pharmacokinetics, the AUC of MTX was enhanced by approx. 6.15 times and plasma half-life was enhanced by 2.45 times, after parenteral administration of single equivalent dose in rodents. FLU-MTX offered enhanced availability of drug to the biological system, meanwhile improved the cancer-cell cytotoxicity, sustained the effective plasma drug concentrations, and offered substantial compatibility to erythrocytes.


breast cancer hydroxylated fullerene chemotherapy drug delivery nanoparticle antifolate drug 



University Grant Commission, New Delhi, India, is highly acknowledged for Rajeev Gandhi National Fellowship to Mr. Manish Kumar (F1-17.1/2014-15/RGNF-2014-15 ST-RAJ-73879/(SA-III/Website)). This work would have been impossible without the gift sample of MTX from M/s Ipca Laboratories, Mumbai, India. Their support for the academic cause is highly appreciated.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflicts of interest.


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Copyright information

© American Association of Pharmaceutical Scientists 2017

Authors and Affiliations

  • Shradha Bahuguna
    • 1
  • Manish Kumar
    • 1
  • Gajanand Sharma
    • 2
  • Rajendra Kumar
    • 3
  • Bhupinder Singh
    • 2
    • 3
  • Kaisar Raza
    • 1
  1. 1.Department of Pharmacy, School of Chemical Sciences and PharmacyCentral University of RajasthanAjmerIndia
  2. 2.Division of Pharmaceutics, University Institute of Pharmaceutical Sciences, Centre of Advanced StudiesPanjab UniversityChandigarhIndia
  3. 3.UGC-Centre of Excellence in Applications of Nanomaterials, Nanoparticles and NanocompositesPanjab UniversityChandigarhIndia

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