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Development of High-Content Gemcitabine PEGylated Liposomes and Their Cytotoxicity on Drug-Resistant Pancreatic Tumour Cells

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ABSTRACT

Purpose

The objective of this study was to develop high-content gemcitabine PEGylated liposomes to reverse gemcitabine resistance in pancreatic tumour cells. The mechanism of drug loading into liposomes was also investigated.

Methods

To increase the drug entrapment efficiency (EE) and drug loading (DL), a novel passive loading approach named Small Volume Incubation method (SVI) was developed and compared to the reverse phase evaporation (REV) and remote loading methods. The in vitro cytotoxicity was evaluated using MIA PaCa-2 and Panc-1 cell lines.

Results

The EE for remote loading was 12.3 ± 0.3%, much lower than expected and a burst release was observed with the resultant liposomes. Using the optimized SVI method, increased EE (37 ± 1%) and DL (4%, w/w) were obtained. The liposomes (200 ± 5 nm) showed minimal drug leakage, good stability, and significant improvement in cytotoxicity to the gemcitabine-resistant pancreatic cancer cell lines.

Conclusions

Remote loading was not suitable for loading gemcitabine into liposomes. pKa > 4.6 for basic drugs and intra-liposomal precipitation of loaded compounds were suggested as an additional requirement to the current criteria for remote loading using ammonium sulphate gradient (pKa < 11). High DL is essential for liposomes to reverse gemcitabine resistance in pancreatic cell lines.

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Abbreviations

DL:

Drug loading

EE:

Entrapment efficiency

MDR:

Multidrug resistance

MTT:

3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide

PEG:

Polyethylene glycol

REV:

Reverse phase evaporation

SVI:

Small volume incubation

TFH:

Thin film hydration

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ACKNOWLEDGMENTS AND DISCLOSURES

The financial support for this study was provided by New Zealand Cancer Society (Grant Number 3627223). The authors also wish to acknowledge the support of a Doctorial Scholarship for Hongtao Xu provided by The University of Auckland, New Zealand. The authors declare that they have no conflicts of interest to disclose.

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Authors and Affiliations

  1. School of Pharmacy, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand

    Hongtao Xu, Wenli Zhang, Linda Duxfield & Zimei Wu

  2. Department of Pharmacology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand

    James Paxton & Joanne Lim

  3. Auckland University of Technology, Auckland, New Zealand

    Yan Li

  4. School of Pharmacy, University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand

    Zimei Wu

Authors
  1. Hongtao Xu
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  2. James Paxton
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  3. Joanne Lim
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  4. Yan Li
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  5. Wenli Zhang
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Correspondence to Zimei Wu.

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Xu, H., Paxton, J., Lim, J. et al. Development of High-Content Gemcitabine PEGylated Liposomes and Their Cytotoxicity on Drug-Resistant Pancreatic Tumour Cells. Pharm Res 31, 2583–2592 (2014). https://doi.org/10.1007/s11095-014-1353-z

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  • DOI: https://doi.org/10.1007/s11095-014-1353-z

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