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GE11-modified liposomes for non-small cell lung cancer targeting: preparation, ex vitro and in vivo evaluation.

Cheng L, Huang FZ, Cheng LF, Zhu YQ, Hu Q, Li L, Wei L, Chen DW - Int J Nanomedicine (2014)

Bottom Line: Interestingly, the cytotoxic effect of the liposomes on A549 tumor cells was closely related to GE11 density, and liposomes with 10% GE11 had the highest tumor cell killing activity and a 2.6-fold lower half maximal inhibitory concentration than that of the nontargeted counterpart (PEG-LP/DOX).Fluorescence microscopy and flow cytometry analysis revealed that GE11 significantly increased cellular uptake of the liposomes, which could be ascribed to specific EGFR-mediated endocytosis.It was found that multiple endocytic pathways were involved in entry of GE11-LP/DOX into cells, but GE11 assisted in cellular internalization mainly via the clathrin-mediated endocytosis pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou, Jiangsu Province, People's Republic of China.

ABSTRACT
Non-small cell lung cancer (NSCLC) is a serious threat to human health, and 40%-80% of NSCLCs express high levels of epidermal growth factor receptor (EGFR). GE11 is a novel peptide and exhibits high affinity for EGFR binding. The aim of this study was to construct and evaluate GE11-modified liposomes for targeted drug delivery to EGFR-positive NSCLC. Doxorubicin, a broad-spectrum antitumor agent, was chosen as the payload. GE11 was conjugated to the distal end of DSPE-PEG2000-Mal by an addition reaction with a conjugation efficiency above 90%. Doxorubicin-loaded liposomes containing GE11 (GE11-LP/DOX) at densities ranging from 0% to 15% were prepared by combination of a thin film hydration method and a post insertion method. Irrespective of GE11 density, the physicochemical properties of these targeted liposomes, including particle size, zeta potential, and drug entrapment efficiency, were nearly identical. Interestingly, the cytotoxic effect of the liposomes on A549 tumor cells was closely related to GE11 density, and liposomes with 10% GE11 had the highest tumor cell killing activity and a 2.6-fold lower half maximal inhibitory concentration than that of the nontargeted counterpart (PEG-LP/DOX). Fluorescence microscopy and flow cytometry analysis revealed that GE11 significantly increased cellular uptake of the liposomes, which could be ascribed to specific EGFR-mediated endocytosis. It was found that multiple endocytic pathways were involved in entry of GE11-LP/DOX into cells, but GE11 assisted in cellular internalization mainly via the clathrin-mediated endocytosis pathway. Importantly, the GE11-modified liposomes showed enhanced accumulation and prolonged retention in tumor tissue, as evidenced by a 2.2-fold stronger mean fluorescence intensity in tumor tissue than the unmodified liposomes at 24 hours. In summary, GE11-modified liposomes may be a promising platform for targeted delivery of chemotherapeutic drugs in NSCLC.

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Related in: MedlinePlus

1H-NMR (A) and Fourier transform infrared spectra (B) for DSPE-PEG2000-GE11 and high-pressure liquid chromatographic curves for DSPE-PEG2000-Mal (C-a), free GE11 (C-b), DTT (C-c), the reaction mixture (C-d), and the reaction mixture treated with DTT (C-e).Abbreviations: DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; PEG, polyethylene glycol; DTT, DL-dithiothreitol; DMSO, dimethyl sulfoxide.
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f1-ijn-9-921: 1H-NMR (A) and Fourier transform infrared spectra (B) for DSPE-PEG2000-GE11 and high-pressure liquid chromatographic curves for DSPE-PEG2000-Mal (C-a), free GE11 (C-b), DTT (C-c), the reaction mixture (C-d), and the reaction mixture treated with DTT (C-e).Abbreviations: DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; PEG, polyethylene glycol; DTT, DL-dithiothreitol; DMSO, dimethyl sulfoxide.

Mentions: GE11 was conjugated to DSPE-PEG2000-Mal by an addition reaction and the structure of the product was verified by 1H-NMR and Fourier transform infrared spectroscopy. As shown in Figure 1A, the peaks at 0.81 ppm and 1.23 ppm were attributed to methyl and methylene protons in DSPE, while the peaks at 3.53 ppm and 6.5–9.3 ppm were attributed to PEG and GE11, respectively. In the Fourier transform infrared spectrum (Figure 1B), the peak at 1,105 cm−1 was the stretching vibration of C–O–C in the PEG segment and the peak at 1,679 cm−1 was the stretching vibration of C(=O)–N in GE11. These results suggested that GE11 was successfully conjugated to DSPE-PEG2000-Mal.


GE11-modified liposomes for non-small cell lung cancer targeting: preparation, ex vitro and in vivo evaluation.

Cheng L, Huang FZ, Cheng LF, Zhu YQ, Hu Q, Li L, Wei L, Chen DW - Int J Nanomedicine (2014)

1H-NMR (A) and Fourier transform infrared spectra (B) for DSPE-PEG2000-GE11 and high-pressure liquid chromatographic curves for DSPE-PEG2000-Mal (C-a), free GE11 (C-b), DTT (C-c), the reaction mixture (C-d), and the reaction mixture treated with DTT (C-e).Abbreviations: DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; PEG, polyethylene glycol; DTT, DL-dithiothreitol; DMSO, dimethyl sulfoxide.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928463&req=5

f1-ijn-9-921: 1H-NMR (A) and Fourier transform infrared spectra (B) for DSPE-PEG2000-GE11 and high-pressure liquid chromatographic curves for DSPE-PEG2000-Mal (C-a), free GE11 (C-b), DTT (C-c), the reaction mixture (C-d), and the reaction mixture treated with DTT (C-e).Abbreviations: DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine; PEG, polyethylene glycol; DTT, DL-dithiothreitol; DMSO, dimethyl sulfoxide.
Mentions: GE11 was conjugated to DSPE-PEG2000-Mal by an addition reaction and the structure of the product was verified by 1H-NMR and Fourier transform infrared spectroscopy. As shown in Figure 1A, the peaks at 0.81 ppm and 1.23 ppm were attributed to methyl and methylene protons in DSPE, while the peaks at 3.53 ppm and 6.5–9.3 ppm were attributed to PEG and GE11, respectively. In the Fourier transform infrared spectrum (Figure 1B), the peak at 1,105 cm−1 was the stretching vibration of C–O–C in the PEG segment and the peak at 1,679 cm−1 was the stretching vibration of C(=O)–N in GE11. These results suggested that GE11 was successfully conjugated to DSPE-PEG2000-Mal.

Bottom Line: Interestingly, the cytotoxic effect of the liposomes on A549 tumor cells was closely related to GE11 density, and liposomes with 10% GE11 had the highest tumor cell killing activity and a 2.6-fold lower half maximal inhibitory concentration than that of the nontargeted counterpart (PEG-LP/DOX).Fluorescence microscopy and flow cytometry analysis revealed that GE11 significantly increased cellular uptake of the liposomes, which could be ascribed to specific EGFR-mediated endocytosis.It was found that multiple endocytic pathways were involved in entry of GE11-LP/DOX into cells, but GE11 assisted in cellular internalization mainly via the clathrin-mediated endocytosis pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou, Jiangsu Province, People's Republic of China.

ABSTRACT
Non-small cell lung cancer (NSCLC) is a serious threat to human health, and 40%-80% of NSCLCs express high levels of epidermal growth factor receptor (EGFR). GE11 is a novel peptide and exhibits high affinity for EGFR binding. The aim of this study was to construct and evaluate GE11-modified liposomes for targeted drug delivery to EGFR-positive NSCLC. Doxorubicin, a broad-spectrum antitumor agent, was chosen as the payload. GE11 was conjugated to the distal end of DSPE-PEG2000-Mal by an addition reaction with a conjugation efficiency above 90%. Doxorubicin-loaded liposomes containing GE11 (GE11-LP/DOX) at densities ranging from 0% to 15% were prepared by combination of a thin film hydration method and a post insertion method. Irrespective of GE11 density, the physicochemical properties of these targeted liposomes, including particle size, zeta potential, and drug entrapment efficiency, were nearly identical. Interestingly, the cytotoxic effect of the liposomes on A549 tumor cells was closely related to GE11 density, and liposomes with 10% GE11 had the highest tumor cell killing activity and a 2.6-fold lower half maximal inhibitory concentration than that of the nontargeted counterpart (PEG-LP/DOX). Fluorescence microscopy and flow cytometry analysis revealed that GE11 significantly increased cellular uptake of the liposomes, which could be ascribed to specific EGFR-mediated endocytosis. It was found that multiple endocytic pathways were involved in entry of GE11-LP/DOX into cells, but GE11 assisted in cellular internalization mainly via the clathrin-mediated endocytosis pathway. Importantly, the GE11-modified liposomes showed enhanced accumulation and prolonged retention in tumor tissue, as evidenced by a 2.2-fold stronger mean fluorescence intensity in tumor tissue than the unmodified liposomes at 24 hours. In summary, GE11-modified liposomes may be a promising platform for targeted delivery of chemotherapeutic drugs in NSCLC.

Show MeSH
Related in: MedlinePlus