Pharmaceutical Research

, Volume 19, Issue 3, pp 265–269

Use of the Post-Insertion Technique to Insert Peptide Ligands into Pre-Formed Stealth Liposomes with Retention of Binding Activity and Cytotoxicity

  • João N. Moreira
  • Tatsuhiro Ishida
  • Rogério Gaspar
  • Theresa M. Allen
Article

DOI: 10.1023/A:1014434732752

Cite this article as:
Moreira, J.N., Ishida, T., Gaspar, R. et al. Pharm Res (2002) 19: 265. doi:10.1023/A:1014434732752

Abstract

Purpose: Simple methods for the large-scale manufacture of ligand-targeted liposomes will be needed if clinical trials are to proceed. We tested a recently developed technology for inserting peptide ligands into preformed Stealth liposomes. Antagonist G-targeted liposomes (PLG) were prepared and loaded with doxorubicin and their cellular association and cytotoxicity were evaluated using the human small cell lung cancer H69 cell line.

Methods: The hexapeptide antagonist G was covalently coupled via a thioether bond to the terminus of polyethylene glycol (PEG) in micelles formed from maleimide-derivatized poly(ethylene glycol) (Mr 2000) distearoylphosphatidylethanolamine followed by transfer into preformed liposomes during a one-step incubation. For cellular association, we used radiolabeled liposomes. Cytotoxicity was evaluated using the MTT in vitro proliferation assay.

Results: The postinsertion approach to the formation of peptide-targeted liposomes led to the production of PLG bearing a maximum of approximately 0.3 μg antagonist G/μmol phospholipid. These liposomes had increased cellular association to H69 cells relative to nontargeted liposomes and, when loaded with doxorubicin, they resulted in similar levels of cytotoxicity to those obtained by conventional coupling techniques.

Conclusions: The postinsertion technique is a simple, effective means for the production of biologically active peptide-targeted liposomes.

antagonist G polyethylene glycol targeting liposomes doxorubicin small cell lung cancer 

Copyright information

© Plenum Publishing Corporation 2002

Authors and Affiliations

  • João N. Moreira
    • 1
    • 2
  • Tatsuhiro Ishida
    • 3
  • Rogério Gaspar
    • 2
  • Theresa M. Allen
    • 3
  1. 1.Department of PharmacologyUniversity of AlbertaEdmontonCanada
  2. 2.Faculty of Pharmacy and Center for NeurosciencesUniversity of CoimbraCoimbraPortugal
  3. 3.Alliance Pharmaceutical Corp.San Diego

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