Protein macromonomers containing reduction-sensitive linkers for covalent immobilization and glutathione triggered release from dextran hydrogels

Verheyen, Ellen; Wal, Steffen van der; Deschout, Hendrik; Braeckmans, Kevin; Smedt, Stefaan C. De; Barendregt, Arjan; Hennink, Wim E.; Nostrum, Cornelus F. van

doi:10.1016/j.jconrel.2011.08.040

Cited by 30 publications

(25 citation statements)

References 55 publications

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“…In the body, the polymer chain may either hydrolytically degrade off of the conjugates, or reductively cleave prior to hydrolytic degradation 5,42 . To visualize the reductive cleavage of the polymer chain from Lyz, the conjugates were incubated with 0.65 M DTT at 95 °C for 6 minutes (Scheme 2).…”

Section: Resultsmentioning

confidence: 99%

Degradable PEGylated protein conjugates utilizing RAFT polymerization

Decker

Maynard

2015

European Polymer Journal

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Poly(ethylene glycol) (PEG)-protein therapeutics exhibit enhanced pharmacokinetics, but have drawbacks including decreased protein activities and polymer accumulation in the body. Therefore a major aim for second-generation polymer therapeutics is to introduce degradability into the backbone. Herein we describe the synthesis of poly(poly(ethylene glycol methyl ether methacrylate)) (pPEGMA) degradable polymers with protein-reactive end-groups via reversible addition-fragmentation chain transfer (RAFT) polymerization, and the subsequent covalent attachment to lysozyme through a reducible disulfide linkage. RAFT copolymerization of cyclic ketene acetal (CKA) monomer 5,6-benzo-2-methylene-1,3-dioxepane (BMDO) with PEGMA yielded two polymers with number-average molecular weight (Mn) (GPC) of 10.9 and 20.9 kDa and molecular weight dispersities (Ð) of 1.34 and 1.71, respectively. Hydrolytic degradation of the polymers was analyzed by 1H-NMR and GPC under basic and acidic conditions. The reversible covalent attachment of these polymers to lysozyme, as well as the hydrolytic and reductive cleavage of the polymer from the protein, was analyzed by gel electrophoresis and mass spectrometry. Following reductive cleavage of the polymer, an increase in activity was observed for both conjugates, with the released protein having full activity. This represents a method to prepare PEGylated proteins, where the polymer is readily cleaved from the protein and the main chain of the polymer is degradable.

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Section: Resultsmentioning

confidence: 99%

Degradable PEGylated protein conjugates utilizing RAFT polymerization

Decker

Maynard

2015

European Polymer Journal

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“…The diffusion mechanism of solute molecules within hydrogels is historically of great interest for a wide variety of industrial applications, such as separations through membranes1–4 and chromatographic columns 5. 6 Moreover, in recent years, diffusion studies applied to nanomedicine and, in particular, to hydrogels that are able to control and sustain the release of drugs have played a leading role 7–10. Indeed several reports and reviews written recently, underline the importance of hydrogels in this field 11–14.…”

Section: Introductionmentioning

confidence: 99%

Drug–Polymer Interactions in Hydrogel‐based Drug‐Delivery Systems: An Experimental and Theoretical Study

et al. 2015

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In drug-delivery systems, drug transport is a key step, but the interpretation of the transport mechanism is still controversial. Here, we investigated a promising hydrogel library loaded with the anticonvulsant drug ethosuximide (ESM). The self-diffusion coefficient of ESM was measured using two methods: a direct and advanced measurement with a pulsed field gradient spin-echo (PFGSE) method, using an NMR spectrometer equipped with high-resolution magic angle spinning (HR-MAS) probe, and an indirect one based on fitting in vitro drug-delivery data. Starting from the experimental data a mathematical model without fitted parameters was developed and all the phenomena involved, that is, adsorption and diffusion, were considered. At low drug concentrations, adsorption prevails and consequently the diffusivity in the gels is lower than that in water. At high drug concentrations, where all adsorption sites are saturated, the diffusion in the gels is similar to that in a water solution. This study may pave the way for better device design.

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“…The addition of a radical initiator increased the degree of protein modification compared to mPEG5k‐acrylamide alone (Figure D). Side reactions between proteins and acrylate‐containing polymers during radical polymerization of hydrogels have been described by several groups . For example, incomplete protein release and loss of activity were observed, most likely due to covalent binding of proteins through radical chain transfer to thiols or other susceptible groups.…”

Section: Discussionmentioning

confidence: 99%

Protein Compatibility of Selected Cross‐linking Reactions for Hydrogels

Hammer

Brandl

Kirchhof

et al. 2014

Macromolecular Bioscience

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The compatibility of selected cross-linking reactions with lysozyme is investigated. Michael-type additions of nucleophilic amino acids to maleimide, vinyl sulfone and acrylamide groups are detected by gel electrophoresis. The degree of modification depends on the polymer and the pH. Complete modification with more than five PEG chains is observed after incubation with mPEG5k-vinyl sulfone at pH 9, whereas 96% of the protein remains unmodified after incubation with mPEG5k-acrylamide at pH 4. Incubation with mPEG5k-thiol results in thiol-disulfide exchange reactions. Hydrogel preparation is simulated by using polymer mixtures. Protein modifications are detected, which may affect the protein structure, decrease activity and bioavailability, and increase the risk for immune responses.

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Protein macromonomers containing reduction-sensitive linkers for covalent immobilization and glutathione triggered release from dextran hydrogels

Cited by 30 publications

References 55 publications

Degradable PEGylated protein conjugates utilizing RAFT polymerization

Degradable PEGylated protein conjugates utilizing RAFT polymerization

Drug–Polymer Interactions in Hydrogel‐based Drug‐Delivery Systems: An Experimental and Theoretical Study

Protein Compatibility of Selected Cross‐linking Reactions for Hydrogels

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