Pharmaceutical nanotechnologyPeptide ligand-mediated liposome distribution and targeting to EGFR expressing tumor in vivo
Introduction
Liposomes have been increasingly developed as preferred drug carriers in anti-tumor treatments (Bangham et al., 1965, Bangham, 1968). PEG-conjugated liposomes can somewhat escape from the capture of reticuloendothelial system to circulate longer than conventional liposomes. So they have a tendency to traverse leaky vascular and passively accumulate in tumor tissues based on the so-called enhanced permeability and retention (EPR) effect (Allen et al., 1991, Lasic et al., 1991, Woodle and Lasic, 1992, Wu et al., 1993). Such stealth liposome formulations had been used to deliver anti-cancer drugs and attained significant therapy effect (Mayhew et al., 1992, Vaage et al., 1992). In addition, modification of liposomes with antibody or antibody fragment, or small molecular ligands has also been developed a promising strategy for tumor targeting. Drug-loaded liposomes with active targeting were shown to have more enhanced anti-cancer efficacy (Lopes de Menezes et al., 1998, Asai et al., 2002, Gabizon et al., 2003).
The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase over-expressed on many human cancer cells surface. It is regarded as a significant target for tumor-targeted therapy. EGFR-targeted immunoliposomes were shown to promote efficient intracellular delivery of doxorubicin to tumor cells, and therefore resulted in superior anti-tumor effects in a series of animal xenograft models (Mamot et al., 2003, Mamot et al., 2005, Mamot et al., 2006). In addition to antibodies and antibody fragments, peptide ligands having specific interaction with receptors over-expressing in tumor or tumor neovasculature were utilized to direct chemotherapeutics, proteins, and gene constructs (Kok et al., 2002). Some peptides such as the RGD-derived peptides were widely used (Takikawa et al., 2000, Schiffelers et al., 2003, Maeda et al., 2004). Many other studies had attempted to find other effective ligands for various targets based on phage display screening, comparative sequence/structure analysis and so on (Morpurgo et al., 2002, Wu et al., 2004). We had previously reported the screening of a noval peptide ligand (GE11) for EGFR (Li et al., 2005). It was shown to bind effectively to EGFR over-expressing cancer cells-mediated target specific gene transfection when conjugated to a PEI vector.
In this study, we used this peptide ligand to construct an active targeting liposome drug delivery system towards EGFR positive cancer cells. We also examined the liposome distribution in vivo in tumor bearing mice. The aim of this study is to explore the feasibility of using peptide ligand-directed liposome as target therapeutics for cancer.
Section snippets
Materials
The sequence of the peptide GE11 is YHWYGYTPQNVI as described in reference Li et al. (2005). It was custom synthesized by GL Biochem Ltd. (shanghai). Its structure and purity were confirmed by HPLC and MS. The irrelated peptide D11 is also synthesized for control. Recombinant hEGF was a gift from Dr. Li Z.P. (State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences).
The lipid 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) was
Preparation of peptide ligand-conjugated liposomes
All the liposomes were prepared and sized by extrusion through 100 nm membranes. The ligand-conjugated DSPE–PEG2000 molecules were then inserted into preformed liposomes based on the method developed by Ishida et al. (Ishida et al., 1999). The process had little disturbance to the integrity of the liposomes. The liposome size distribution before and after the insertion were shown in Table 1.
Binding and endocytosis of ligand-modified liposomes by EGFR expressing cells
Both EGF- and GE11-modified liposomes and control mPEG–DSPE liposomes were added to cultured EGFR
Discussion
For targeted drug delivery towards tumor, small molecule ligands, peptides, antibody and antibody fragments had all been used (Ahmad et al., 1993, Woodle et al., 2001, Kok et al., 2002, Gabizon et al., 2003, Schiffelers et al., 2003, Pastorino et al., 2006, Saul et al., 2006, Oba et al., 2007). EGFR is an important target for its over-expressed in many cancers (Schmidt et al., 1997, Lutsenko et al., 2002, Mamot et al., 2003). In our previous study (Li et al., 2005), peptide GE11 was identified
Acknowledgements
Grant support: National Science Foundation of China Grant No. 30472097 and Chinese National 973 Grant No. 2004CB518802
References (43)
- et al.
Liposomes containing synthetic lipid derivatives of poly(ethylene glycol) show prolonged circulation half-lives in vivo
Biochim. Biophys. Acta
(1991) - et al.
Anti-neovascular therapy by liposomal DPP–CNDAC targeted to angiogenic vessels
FEBS Lett.
(2002) Membrane models with phospholipids
Prog. Biophys. Mol. Biol.
(1968)- et al.
Attachment of antibodies to sterically stabilized liposomes: evaluation, comparison and optimization of coupling procedures
Biochim. Biophys. Acta
(1995) - et al.
Transmembrane ammonium sulfate gradients in liposomes produce efficient and stable entrapment of amphipathic weak bases
Biochim. Biophys. Acta
(1993) - et al.
In vitro and in vivo comparison of immunoliposomes made by conventional coupling techniques with those made by a new post-insertion approach
Biochim. Biophys. Acta
(2001) - et al.
A combinatorial approach to producing sterically stabilized (Stealth) immunoliposomal drugs
FEBS Lett.
(1999) - et al.
Sterically stabilized liposomes: a hypothesis on the molecular origin of the extended circulation times
Biochim. Biophys. Acta
(1991) - et al.
Anti-neovascular therapy by use of tumor neovasculature-targeted long-circulating liposome
J. Control. Release
(2004) - et al.
Targetability of novel immunoliposomes modified with amphipathic poly(ethylene glycol)s conjugated at their distal terminals to monoclonal antibodies
Biochim. Biophys. Acta
(1995)
Characterization of liposomal systems containing doxorubicin entrapped in response to pH gradients
Biochim. Biophys. Acta
An approach to increased polyplex gene delivery by peptides selected from a phage display library
J. Biochem. Biophys. Methods
Chemistry for peptide and protein PEGylation
Adv. Drug Deliv. Rev.
A dual-ligand approach for enhancing targeting selectivity of therapeutic nanocarriers
J. Control. Release
Anti-tumor efficacy of tumor vasculature-targeted liposomal doxorubicin
J. Control. Release
Suppression of GD1alpha ganglioside-mediated tumor metastasis by liposomalized WHW-peptide
FEBS Lett.
Tumor targeting based on the effect of enhanced permeability and retention (EPR) and the mechanism of receptor-mediated endocytosis (RME)
Int. J. Pharm.
Sterically stabilized liposomes
Biochim. Biophys. Acta
Sterically stabilized polyplex: ligand-mediated activity
J. Control. Release
A novel small peptide as a targeting ligand for receptor tyrosine kinase Tie2
Biochem. Biophys. Res. Commun.
Antibody-targeted delivery of doxorubicin entrapped in sterically stabilized liposomes can eradicate lung cancer in mice
Cancer Res.
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2022, European Journal of Pharmaceutics and BiopharmaceuticsCitation Excerpt :While with SPR measurements, ligands and receptors interact in monomolecular form, binding studies on cells involve all receptors on the cell surface, and potential positioning effects, interaction with other cell surface components, but also internalization can add up to the binding affinity. While EGFR-targeting peptides exhibit far lower affinity for EGFR compared to EGF and might not lead to significant targeting when used in monomolecular settings, their successful use as targeting ligand for macromolecular drug—and nucleic acid delivery systems is undoubted [8,9,14,15,17,19,21,24,38]. We and others have used polycations like linear polyethylenimine (LPEI) for gene delivery purposes in vitro and in vivo [28,39].