Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release

Background and Aim: Polyrotaxanes are molecular necklaces composed of polymers, threaded macrocycles, and bulky stopper molecules to inhibit the decomposition of the first two. These supramolecular assemblies are promising active ingredients for treating lysosomal storage disorders. This study aimed...

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Main Authors: Gergely Kali, Alexander H. Mayer, Dennis To, Martyna Truszkowska, Raphael Plangger, Markus Gallei, Andreas Bernkop-Schnürch
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Carbohydrate Polymer Technologies and Applications
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Online Access:http://www.sciencedirect.com/science/article/pii/S266689392400166X
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author Gergely Kali
Alexander H. Mayer
Dennis To
Martyna Truszkowska
Raphael Plangger
Markus Gallei
Andreas Bernkop-Schnürch
author_facet Gergely Kali
Alexander H. Mayer
Dennis To
Martyna Truszkowska
Raphael Plangger
Markus Gallei
Andreas Bernkop-Schnürch
author_sort Gergely Kali
collection DOAJ
description Background and Aim: Polyrotaxanes are molecular necklaces composed of polymers, threaded macrocycles, and bulky stopper molecules to inhibit the decomposition of the first two. These supramolecular assemblies are promising active ingredients for treating lysosomal storage disorders. This study aimed to synthesize such polyrotaxanes with a novel, simple method, resulting in high threading efficacy, enhanced cellular uptake, and intracellular cyclodextrin (CD) release. Methods: In this study, we developed two novel poly(ethylene glycol) (PEG) based polyrotaxanes, with threaded α-cyclodextrin (α-CD) or its mixture with 2-hydroxypropyl-α-CD (HP-α-CD) and glutathione sensitive disulfide connected stopper molecules. The structure and composition of these polyrotaxanes were determined by 1H NMR spectroscopy and gel permeation chromatography, while the cellular uptake was investigated by flow cytometry and confocal microscopy. Results: High threading efficacy, as well as molar mass of 17.9 and 13.1 kDa, was found for the polymeric supramolecules with threaded α-CD and α-CD/HP-α-CD, respectively. Glutathion-triggered reductive removal of the stopper molecules showed potential decomposition of these polyrotaxanes in target cells. Flow cytometry revealed an up to 52-fold enhancement in cellular uptake of α- and HP-α-CD by the polyrotaxanes compared to free CD, which was also visualized by confocal microscopy. Conclusion and scope of application: Polyrotaxanes based on α-CD and its derivative were tested in vitro for application in the treatment of lysosomal storage disease for the first time. Based on these results, polyrotaxanes with disulfide stopper molecules might be promising supramolecular excipients for cellular delivery of α-CDs.
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publishDate 2024-12-01
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spelling doaj-art-be7cf216490c48c0b285eb97c6bed3cc2024-12-13T11:06:32ZengElsevierCarbohydrate Polymer Technologies and Applications2666-89392024-12-018100586Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin releaseGergely Kali0Alexander H. Mayer1Dennis To2Martyna Truszkowska3Raphael Plangger4Markus Gallei5Andreas Bernkop-Schnürch6Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, AustriaCenter for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, AustriaCenter for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, AustriaCenter for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, AustriaCenter for Chemistry and Biomedicine, Department of Organic Chemistry, Institute of Chemistry, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, AustriaPolymer Chemistry, Saarland University, Campus C4 2, 66123 Saarbrücken, Germany; Saarene, Saarland Center for Energy Materials and Sustainability, Campus C4 2, 66123 Saarbrücken, GermanyCenter for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria; Corresponding author at: Center for Chemistry and Biomedicine, Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.Background and Aim: Polyrotaxanes are molecular necklaces composed of polymers, threaded macrocycles, and bulky stopper molecules to inhibit the decomposition of the first two. These supramolecular assemblies are promising active ingredients for treating lysosomal storage disorders. This study aimed to synthesize such polyrotaxanes with a novel, simple method, resulting in high threading efficacy, enhanced cellular uptake, and intracellular cyclodextrin (CD) release. Methods: In this study, we developed two novel poly(ethylene glycol) (PEG) based polyrotaxanes, with threaded α-cyclodextrin (α-CD) or its mixture with 2-hydroxypropyl-α-CD (HP-α-CD) and glutathione sensitive disulfide connected stopper molecules. The structure and composition of these polyrotaxanes were determined by 1H NMR spectroscopy and gel permeation chromatography, while the cellular uptake was investigated by flow cytometry and confocal microscopy. Results: High threading efficacy, as well as molar mass of 17.9 and 13.1 kDa, was found for the polymeric supramolecules with threaded α-CD and α-CD/HP-α-CD, respectively. Glutathion-triggered reductive removal of the stopper molecules showed potential decomposition of these polyrotaxanes in target cells. Flow cytometry revealed an up to 52-fold enhancement in cellular uptake of α- and HP-α-CD by the polyrotaxanes compared to free CD, which was also visualized by confocal microscopy. Conclusion and scope of application: Polyrotaxanes based on α-CD and its derivative were tested in vitro for application in the treatment of lysosomal storage disease for the first time. Based on these results, polyrotaxanes with disulfide stopper molecules might be promising supramolecular excipients for cellular delivery of α-CDs.http://www.sciencedirect.com/science/article/pii/S266689392400166XCyclodextrinPoly(ethylene glycol)PolyrotaxaneCellular uptakeDisulfide stopper molecules
spellingShingle Gergely Kali
Alexander H. Mayer
Dennis To
Martyna Truszkowska
Raphael Plangger
Markus Gallei
Andreas Bernkop-Schnürch
Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
Carbohydrate Polymer Technologies and Applications
Cyclodextrin
Poly(ethylene glycol)
Polyrotaxane
Cellular uptake
Disulfide stopper molecules
title Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
title_full Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
title_fullStr Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
title_full_unstemmed Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
title_short Disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
title_sort disulfide stoppered polyrotaxanes with enhanced cellular uptake and intracellular cyclodextrin release
topic Cyclodextrin
Poly(ethylene glycol)
Polyrotaxane
Cellular uptake
Disulfide stopper molecules
url http://www.sciencedirect.com/science/article/pii/S266689392400166X
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