Performance of cell-penetrating peptide-linked polymers physically mixed with poorly membrane-permeable molecules on cell membranes
| Autor: | Atsushi Kasai, Yuki Ishimaru, Ryoji Kimura, Akio Hashizume, Shinji Yamashita, Yoshie Masaoka, Noriko Nakajima, Sadaaki Maeda, Ken-ichiro Hiwatari, Hiroyuki Tachikawa, Hitoshi Yamauchi, Shinji Sakuma, Takafumi Yamamoto, Masaya Suita, Makoto Kataoka, Norihiro Shinkai |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Cell Membrane Permeability
Time Factors Membrane permeability Cell Pharmaceutical Science Peptide Cell-Penetrating Peptides Rhodamine Cell membrane Amiloride chemistry.chemical_compound Acetamides medicine Humans Fluorescent Dyes chemistry.chemical_classification Microscopy Confocal Rhodamines Pinocytosis Cell Membrane General Medicine Fluoresceins Membrane medicine.anatomical_structure chemistry Biochemistry Acrylates Cell-penetrating peptide Biophysics Polyvinyls Caco-2 Cells Oligopeptides Biotechnology |
| Zdroj: | European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V. 81(1) |
| ISSN: | 1873-3441 |
| Popis: | We are investigating a new class of penetration enhancers that enable poorly membrane-permeable molecules physically mixed with them to effectively penetrate cell membranes without their concomitant cellular uptake. Since we previously revealed that poly(N-vinylacetamide-co-acrylic acid) modified with d -octaarginine, which is a typical cell-penetrating peptide, significantly enhanced the nasal absorption of insulin, we examined the performance of the polymers on cell membranes. When Caco-2 cells were incubated with 5(6)-carboxyfluorescein (CF) for 30 min, approximately 0.1% of applied CF was internalized into the cells. This poor membrane permeability was dramatically enhanced by d -octaarginine-linked polymers; a 25-fold increase in the cellular uptake of CF was observed when the polymer concentration was adjusted to 0.2 mg/mL. None of the individual components, for example, d -octaarginine, had any influence on CF uptake, demonstrating that only d -octaarginine anchored chemically to the polymeric platform enhanced the membrane permeation of CF. The polymer-induced CF uptake was consistently high even when the incubation time was extended to 120 min. Confocal laser scanning microphotographs of cells incubated with d -octaarginine-linked polymers bearing rhodamine red demonstrated that the cell outline was stained with red fluorescence. The polymer-induced CF uptake was significantly suppressed by 5-(N-ethyl-N-isopropyl)amiloride, which is an inhibitor of macropinocytosis. Results indicated that d -octaarginine-linked polymers remained on the cell membrane and poorly membrane-permeable CF was continuously internalized into cells mainly via macropinocytosis repeated for the individual peptidyl branches in the polymer backbone. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect