Selectivity Enhancement of Paclitaxel Liposome Towards Folate Receptor-Positive Tumor Cells by Ligand Number Optimization Approach


The present work aims to develop folate-targeted paclitaxel liposome (F-PTX-LIP), which will selectively target tumor cells overexpressing folate receptor (FR) and leave normal cells. Liposomes were prepared by thin-film hydration method followed by post-insertion of synthesized ligand 1,2-distearoyl-sn-glycero-phosphoethanolamine-polyethyleneglycol 2000-folic acid (DSPE-PEG2000-FA) on the outer surface of the liposome. The synthesized ligand was evaluated for in vivo acute toxicity in Balb/c mice. Developed liposomal formulations were characterized using transmission electron microscopy (TEM) and small-angle neutron scattering (SANS). We have investigated the effect of ligand number on cell uptake and cytotoxicity by confocal laser scanning microscopy (CLSM), competitive inhibition and 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay. Compared to lung adenocarcinoma cells (A549), uptake in human ovarian carcinoma cells (SKOV3) was 2.2- and 1.2-fold higher for liposome with 480 and 240 ligand number respectively. Competitive inhibition experiment shows that prior incubation of SKOV3 cells with free folic acid significantly reduced the cell uptake of F-PTX-LIP with 480 ligand number (480 F-PTX-LIP) by 2.6-fold. 480 F-PTX-LIP displays higher cytotoxicity than free drug and PTX liposome. Moreover, it specifically targets the cells with higher folate receptor expression. Optimized 480 F-PTX-LIP formulation can be potentially useful for the treatment of folate receptor-positive tumors.

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folate receptor




paclitaxel liposome


folate-targeted paclitaxel liposome


1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethyleneglycol 2000


1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethyleneglycol 2000-folic acid


particle size


polydispersity index


zeta potential

% EE:

percent drug entrapment efficiency

% DL:

percent drug loading


transmission electron microscopy


small-angle neutron scattering


differential scanning calorimetry


confocal laser scanning microscopy


3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


FR-targeted liposomes


folate receptor-targeted liposomes with no ligand


blank liposome

240 F-LIP:

folate receptor-targeted liposomes with 240 ligand per liposome

480 F-LIP:

folate receptor-targeted liposomes with 480 ligand per liposome

240 F-PTX-LIP:

folate receptor-targeted paclitaxel liposomes with 240 ligand per liposome

480 F-PTX-LIP:

folate receptor-targeted paclitaxel liposomes with 480 ligand per liposome


polyethylene glycols


soya phosphatidylcholine LIPOID S100










dynamic light scattering


Fourier transform infrared


position-sensitive detector


Roswell Park Memorial Institute


proton nuclear magnetic resonance


mean fluorescence intensity


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The authors would like to acknowledge the Council of Scientific and Industrial Research (CSIR), New Delhi, and Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Mumbai, for providing fellowship. They are also thankful to AICTE-NAFETIC for providing research facilities.

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Correspondence to Pradeep R. Vavia.

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Prajapati, M.K., Bishnu, A., Ray, P. et al. Selectivity Enhancement of Paclitaxel Liposome Towards Folate Receptor-Positive Tumor Cells by Ligand Number Optimization Approach. AAPS PharmSciTech 20, 317 (2019).

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  • nanotechnology
  • liposome
  • paclitaxel
  • DSPE-PEG2000-Folate
  • ligand number
  • folate receptor targeting