Abstract
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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Abbreviations
- FR:
-
folate receptor
- PTX:
-
paclitaxel
- PTX-LIP:
-
paclitaxel liposome
- F-PTX-LIP:
-
folate-targeted paclitaxel liposome
- DSPE-PEG2000:
-
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethyleneglycol 2000
- DSPE-PEG 2000-FA:
-
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethyleneglycol 2000-folic acid
- PS:
-
particle size
- PDI:
-
polydispersity index
- ZP:
-
zeta potential
- % EE:
-
percent drug entrapment efficiency
- % DL:
-
percent drug loading
- TEM:
-
transmission electron microscopy
- SANS:
-
small-angle neutron scattering
- DSC:
-
differential scanning calorimetry
- CLSM:
-
confocal laser scanning microscopy
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- F-LIP:
-
FR-targeted liposomes
- NF-LIP:
-
folate receptor-targeted liposomes with no ligand
- b-LIP:
-
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
- PEG:
-
polyethylene glycols
- SPC:
-
soya phosphatidylcholine LIPOID S100
- DCP:
-
dihexadecylphosphate
- DCC:
-
dicyclohexylcarbodiimide
- NHS:
-
N-hydroxysuccinimide
- TEA:
-
triethylamine
- DLS:
-
dynamic light scattering
- FTIR:
-
Fourier transform infrared
- PSD:
-
position-sensitive detector
- RPMI:
-
Roswell Park Memorial Institute
- 1H NMR:
-
proton nuclear magnetic resonance
- MFI:
-
mean fluorescence intensity
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Acknowledgments
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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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). https://doi.org/10.1208/s12249-019-1531-8
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DOI: https://doi.org/10.1208/s12249-019-1531-8