A Mechanism Enhancing Macromolecule Transport Through Paracellular Spaces Induced by Poly-L-Arginine: Poly-L-Arginine Induces the Internalization of Tight Junction Proteins via Clathrin-Mediated Endocytosis

Yamaki, Tsutomu; Kamiya, Yusuke; Ohtake, Kazuo; Uchida, Masaki; Seki, Toshinobu; Ueda, Hideo; Kobayashi, Jun; Morimoto, Yasunori; Natsume, Hideshi

doi:10.1007/s11095-014-1324-4

Cited by 23 publications

(17 citation statements)

References 27 publications

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“…TJ proteins accumulate in early endosomes as a first station after internalization. Several reports indicate that claudin‐1, claudin‐5, claudin‐16, occludin, JAM‐A, and ZO‐1 accumulate in early endosomes in both epithelial and endothelial cells . Their presence in early endosomes is closely associated with diminished barrier function (hyperpermeability) .…”

Section: Early Endosomes: a Center Of Sortingmentioning

confidence: 99%

“…Several reports indicate that claudin-1, claudin-5, claudin-16, occludin, JAM-A, and ZO-1 accumulate in early endosomes in both epithelial and endothelial cells. 23,25,32,40,48,[68][69][70][71] Their presence in early endosomes is closely associated with diminished barrier function (hyperpermeability). 32,40,47,71 Furthermore, Rab5A depletion using small interfering RNA (siRNA) increased claudin-1 association with the plasma membrane in the human endothelial cell line EA hy.926 and also stabilized brain endothelial barrier function owing to an inability to further process internalized junction proteins.…”

Section: Endocytic Trafficking and Sorting Of Tj Proteinsmentioning

confidence: 99%

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Endocytosis of tight junction proteins and the regulation of degradation and recycling

Stamatovic¹,

Johnson²,

Sladojević

et al. 2017

Annals of the New York Academy of Sciences

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Internalization of tight junction (TJ) proteins from the plasma membrane is a pivotal mechanism regulating TJ plasticity and function in both epithelial and endothelial barrier tissues. Once internalized, the TJ proteins enter complex vesicular machinery, where further trafficking is directly dependent on the initiating stimulus and downstream signaling pathways that regulate the sorting and destiny of TJ proteins, as well as on cell and barrier responses. The destiny of internalized TJ proteins is recycling to the plasma membrane or sorting to late endosomes and degradation. This review highlights recent advances in our knowledge of endocytosis and vesicular trafficking of TJ proteins in both epithelial and endothelial cells. A greater understanding of these processes may allow for the development of methods to modulate barrier permeability for drug delivery or preventing barrier dysfunction in disease states.

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Section: Early Endosomes: a Center Of Sortingmentioning

confidence: 99%

Section: Endocytic Trafficking and Sorting Of Tj Proteinsmentioning

confidence: 99%

Endocytosis of tight junction proteins and the regulation of degradation and recycling

Stamatovic¹,

Johnson²,

Sladojević

et al. 2017

Annals of the New York Academy of Sciences

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“…22) Caco-2 cell sheets were washed with HBSS, and the DMEM culture medium was replaced with HBSS. The test solutions containing polycations (PLO and PEG-PLO at various concentrations) and FD-4 (1.0 mg/mL) were applied to the apical side of the cell sheet.…”

Section: Fd-4 Permeation Experimentsmentioning

confidence: 99%

Preparation and Evaluation of PEGylated Poly-L-ornithine Complex as a Novel Absorption Enhancer

Kamiya

Yamaki

Uchida

et al. 2017

Biological & Pharmaceutical Bulletin

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Polycationic compounds, such as poly-L-arginine and poly-L-ornithine (PLO), enhance the nasal absorption of hydrophilic macromolecular drugs. However, the bio availability corresponding to the dose of these enhancers has not been obtained in an open system study, where an administered solution is transferred to the pharynx because they do not exhibit mucoadhesion/retention in the nasal cavity. In this study, we prepared PEGylated-poly-L-ornithine (PEG-PLO) and investigated the effects of PEGylation on in vitro adhesion/retention properties, permeation enhancement efficiency, and cytotoxicity. PEG-PLO bearing 3-4 polyethylene glycol (PEG) chains per PLO molecule was more retentive than unmodified PLO on an inclined plate. The permeability of a model drug, FD-4, across Caco-2 cell sheets was enhanced by PEG-PLO as well as by PLO. PLO showed cytotoxicity at high concentrations, whereas PEG-PLO did not decrease cell viability, even above the concentration giving a sufficient enhancement effect. These findings suggest that PEGylation of polycationic absorption enhancers improves their adhesion/retention and decreases their cytotoxicity, which may lead to enhancers with greater utility.Key words poly-L-ornithine (PLO); polyethylene glycol (PEG); permeation enhancer Bioactive peptides, such as insulin, teriparatide, liraglutide, and somatropin, have been used widely as therapeutic drugs because of their intrinsic and effective activities, however, they have poor permeability across the epithelium because they are hydrophilic macromolecules. Orally administered peptides are often inactivated in the gastrointestinal tract, and thus peptides are usually administered as injectable formulations. However, injection can result in poor compliance because it is painful and burdensome. 1) Therefore, alternative routes of administration are required.The nasal mucosa has a large surface area for absorption because of its villus structure. Drugs absorbed across the nasal mucosa avoid the hepatic first pass effect because of the abundant vasculature under the nasal mucosa.2) The intranasal route is effective for absorbing high molecular weight drugs 3) and is usually not painful. Therefore, intranasal administration is an important alternative route to injection for peptides and proteins.Polycationic compounds, such as poly-L-arginine (PLA), poly-L-lysine, and poly-L-ornithine (PLO), enhance the transmucosal absorption of hydrophilic macromolecules.4,5) PLA enhances the paracellular permeability of rabbit nasal mucosa and Caco-2 cell sheets reversibly.6,7) The absorption of hydrophilic macromolecules across the nasal mucosa in rats is also increased by co-administration with PLA. 8) PLA enhances the permeability by altering the localization of tight junction proteins from the cell-cell junction to the intracellular space without causing cytotoxicity. 9) Therefore, polycationic absorption enhancers are useful in developing efficient transmucosal drug delivery systems.Nevertheless, it is difficult to increase the bioavailability of dr...

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“…Therefore, it seems that nasal rhGH absorption is increased by increasing the solubility and the paracellular permeability of rhGH by PLA as the same mechanism identified previously. [13][14][15][16][17] With regard to the effects of the molecular weight of PLA, a delay in the rhGH absorption was observed with an increase in the molecular weight. In particular, T max in 1.0 % (w/v) PLA (150) was 9.6 h, and the rhGH absorption was remarkably delayed compared with that in the solutions with other concentrations and molecular weights of PLA.…”

Section: Discussionmentioning

confidence: 99%

“…11,13) In the studies on the mechanism of enhanced absorption by PLA in the mucosa, it has been found that PLA induces the enhanced paracellular permeability by the dispersion of tight junction proteins (claudin-4, occludin, zonula occludens-1 (ZO-1) and tricellulin) into the cytoplasm through the cell-cell junction. [14][15][16][17] In particular, it was known that occludin was important for macromolecular permeation among tight junction proteins. 18,19) In addition, it has been also demonstrated that adherens junction protein (E-cadherin, β-catenin) is dispersed by PLA from the cell-cell junction.…”

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confidence: 99%

Development of a Transnasal Delivery System for Recombinant Human Growth Hormone (rhGH): Effects of the Concentration and Molecular Weight of Poly-L-arginine on the Nasal Absorption of rhGH in Rats

Kawashima

Uchida

Yamaki

et al. 2016

Biological & Pharmaceutical Bulletin

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A novel system for delivering recombinant human growth hormone (rhGH) that is noninvasive and has a simple method of administration is strongly desired to improve the compliance of children. The aim of this study was to investigate the potential for the intranasal (i.n.) co-administration of rhGH with poly-L-arginine (PLA) as a novel delivery system by evaluating the effects of the concentration and molecular weight of PLA on the nasal absorption of rhGH. The influence of the formation of insoluble aggregates and a soluble complex in the dosage formulation on nasal rhGH absorption was also evaluated by size-exclusion chromatography and ultrafiltration. PLA enhanced the nasal absorption of rhGH at each concentration and molecular weight examined. Nasal rhGH absorption increased dramatically when the PLA concentration was 1.0 % (w/v) due to the improved solubility of rhGH in the formulation. A delay in rhGH absorption was observed when the molecular weight of PLA was increased. This appeared to be because the increase in molecular weight caused the formation of a soluble complex. It seems that the PLA concentration affects the absorptionenhancing effect on rhGH, while the molecular weight of PLA affects the time when the maximum plasma rhGH concentration was reached (T max ) of rhGH after i.n. administration, mainly because of the interactions among rhGH, PLA, and additives. Therefore, the transnasal rhGH delivery system using PLA is considered to be a promising alternative to subcutaneous (s.c.) injection if these interactions are sufficiently controlled.Key words poly-L-arginine; nasal absorption; enhancer; recombinant human growth hormone Most peptide and protein medicinal drugs are administered clinically by injection. This invasive administration with a needle and syringe is associated with various problems, such as pain for the patients, the risk of trouble at the injection site, increased medical waste and the potential for complicated operation. Hence, an alternative route of administration is strongly desired. There have been numerous studies on the transmucosal delivery of peptide and protein drugs, such as oral, nasal, and pulmonary administration.1-3) However, these types of drugs are rapidly destroyed by the enzymes present in various organs, especially in the gastrointestinal tract, and have a limited permeation across the mucous membrane. Therefore, these drugs have low bioavailability.Recombinant human growth hormone (rhGH), a single polypeptide chain of 191 amino acids and a molecular mass of 22.1 kDa, is used to treat short stature in children caused by growth hormone deficiency, Turner's syndrome, or chronic renal failure. At present, rhGH is administered daily by subcutaneous (s.c.) injections. Therefore, compliance with rhGH therapy is often low, because children hate the pain caused by such injections. The compliance of patients affects the therapeutic outcome, because it is necessary to regularly administer injections of rhGH for the limited period of growth. 4,5) Hence, a novel rhGH deliver...

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A Mechanism Enhancing Macromolecule Transport Through Paracellular Spaces Induced by Poly-L-Arginine: Poly-L-Arginine Induces the Internalization of Tight Junction Proteins via Clathrin-Mediated Endocytosis

Abstract: PLA induced the transient internalization of TJ proteins in cell-cell junctions via clathrin-mediated endocytosis, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route.

Cited by 23 publications

References 27 publications

Endocytosis of tight junction proteins and the regulation of degradation and recycling

Endocytosis of tight junction proteins and the regulation of degradation and recycling

Preparation and Evaluation of PEGylated Poly-L-ornithine Complex as a Novel Absorption Enhancer

Development of a Transnasal Delivery System for Recombinant Human Growth Hormone (rhGH): Effects of the Concentration and Molecular Weight of Poly-L-arginine on the Nasal Absorption of rhGH in Rats

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