Abstract | BACKGROUND: To facilitate selective drug delivery to hepsin (Hpn)-expressing cancer cells, the RIPL peptide (IPLVVPLRRRRRRRRC; 16mer; 2.1 kDa) was synthesized as a novel cell penetrating/homing peptide (CPHP) and conjugated to a liposomal carrier. METHODS:
RIPL peptide-conjugated liposomes (RIPL-Lipo) were prepared by conjugating RIPL peptides to maleimide-derivatized liposomal vesicles via the thiol-maleimide reaction. Vesicle size and zeta potential were examined using a Zetasizer. Intracellular uptake specificity of the RIPL peptide, or RIPL-Lipo, was assessed by measuring mean fluorescence intensity (MFI) after treatment with a fluorescent marker in various cell lines: SK-OV-3, MCF-7, and LNCaP for Hpn(+); DU145, PC3, and HaCaT for Hpn(-). FITC-dextran was used as a model compound. Selective translocational behavior of RIPL-Lipo to LNCaP cells was visualized by fluorescence microscopy and confocal laser scanning microscopy. Cytotoxicities of the RIPL peptide and RIPL-Lipo were evaluated by WST-1 assay. RESULTS: RIPL peptides exhibited significant Hpn-selectivity. RIPL-Lipo systems were of positively charged nanodispersion (165 nm in average; 6-24 mV depending on RIPL conjugation ratio). RIPL-Lipo with the conjugation of 2300 peptide molecules revealed the greatest MFI in all cell lines tested. Cellular uptake of RIPL-Lipo increased by 20- to 70-fold in Hpn(+) cells, and 5- to 7-fold in Hpn(-) cells, compared to the uptake of FITC-dextran. Cytosolic internalization of RIPL-Lipo was time-dependent: bound instantly; internalized within 30 min; distributed throughout the cytoplasm after 1 h. Cytotoxicities of RIPL peptide (up to 50 μM) and RIPL-Lipo (up to 10%) were minor (cell viability >90%) in LNCaP and HaCaT cells. CONCLUSION: By employing a novel CPHP, the RIPL-Lipo system was successfully developed for Hpn-specific drug delivery.
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Authors | Min Hyung Kang, Min Jung Park, Hyun Joon Yoo, Kwon Yie hyuk, Sang Gon Lee, Sung Rae Kim, Dong Woo Yeom, Myung Joo Kang, Young Wook Choi |
Journal | European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V
(Eur J Pharm Biopharm)
Vol. 87
Issue 3
Pg. 489-99
(Aug 2014)
ISSN: 1873-3441 [Electronic] Netherlands |
PMID | 24704199
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2014 Elsevier B.V. All rights reserved. |
Chemical References |
- Drug Carriers
- Liposomes
- Maleimides
- Peptides
- maleimide
- Serine Endopeptidases
- hepsin
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Topics |
- Cell Line, Tumor
- Drug Carriers
(administration & dosage)
- Drug Delivery Systems
- Humans
- Liposomes
(administration & dosage)
- MCF-7 Cells
- Maleimides
(administration & dosage)
- Microscopy, Fluorescence
(methods)
- Peptides
(administration & dosage)
- Serine Endopeptidases
(metabolism)
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