Analysis of the interactions between Eudragit® L100 and porcine pancreatic trypsin by calorimetric techniques

Braia, Mauricio; Tubío, Gisela; Nerli, Bibiana Beatriz; Loh, Watson; Romanini, Diana

doi:10.1016/j.ijbiomac.2011.10.016

Cited by 13 publications

(8 citation statements)

References 27 publications

(25 reference statements)

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“…56 In addition to electrostatic interactions, their studies on the anionic polyelectrolyte EudragitÒ L100 show that additional hydrophobic groups can lead to signicant contributions. 58 However, several ITC studies indicate that the entropic driving forces for protein adsorption on charged interfaces or polymeric layers is due to counterion release. Welsch et al 59 also showed by ITC that the binding of lysozyme to a negatively charged microgel is an entropically driven process, although evidence for electrostatic interactions also comes from the ionic strength dependence of binding affinity.…”

Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

Protein–polyelectrolyte interactions

et al. 2013

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Section: Isothermal Titration Calorimetry (Itc)mentioning

confidence: 99%

Protein–polyelectrolyte interactions

et al. 2013

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“…EL100-coated PECs partially inhibited insulin from degradation by pancreatin, so the coating seems to perform this role in addition to enabling the pH-dependent release in intestinal regions. EL100 might inactivate trypsin, as the latter seems to bind poly(acrylic acid) [78], while chymotrypsin can bind carboxylate units, leading to enzyme inhibition [79].…”

Section: Discussionmentioning

confidence: 99%

An Enteric-Coated Polyelectrolyte Nanocomplex Delivers Insulin in Rat Intestinal Instillations When Combined with a Permeation Enhancer

et al. 2020

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The use of nanocarriers is being researched to achieve oral peptide delivery. Insulin-associated anionic polyelectrolyte nanoparticle complexes (PECs) were formed that comprised hyaluronic acid and chitosan in an optimum mass mixing ratio of 5:1 (MR 5), followed by coating with a pH-dependent polymer. Free insulin was separated from PECs by size exclusion chromatography and then measured by HPLC. The association efficiency of insulin in PECs was >95% and the loading was ~83 µg/mg particles. Dynamic light scattering and nanoparticle tracking analysis of PECs revealed low polydispersity, a negative zeta potential range of −40 to −50 mV, and a diameter range of 95–200 nm. Dissolution studies in simulated small intestinal fluid (FaSSIF-V2) revealed that the PECs were colloidally stable. PECs that were coated with Eudragit® L-100 delayed insulin release in FaSSIF-V2 and protected insulin against pancreatin attack more than uncoated PECs. Uncoated anionic PECs interacted weakly with mucin in vitro and were non-cytotoxic to Caco-2 cells. The coated and uncoated PECs, both concentrated further by ultrafiltration, permitted dosing of 50 IU/kg in rat jejunal instillations, but they failed to reduce plasma glucose or deliver insulin to the blood. When ad-mixed with the permeation enhancer (PE), sucrose laurate (100 mM), the physicochemical parameters of coated PECs were relatively unchanged, however blood glucose was reduced by 70%. In conclusion, the use of a PE allowed for the PEC-released bioactive insulin to permeate the jejunum. This has implications for the design of orally delivered particles that can release the payload when formulated with enhancers.

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“…Bovine pancreas Trypsin (EC: 3.4.21.4) is a prototype of the serine endopeptidase of S 1 family that consists of a polypeptide chain in which two β-barrel domains are bridged by the catalytic residues 12 .…”

Section: Methodsmentioning

confidence: 99%

The effect of spermine on the structure, thermal stability and activity of bovine pancreatic trypsin

et al. 2016

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This work studied the interaction between spermine and trypsin at pH 8.0. The thermal stability of trypsin was investigated in the presence of spermine over a temperature range (from 293 K to 353 K).Additionally, the conformational change of trypsin induced by spermine was analyzed by UV-Vis absorption and fluorescence spectra. The effect of spermine on trypsin activity was studied at 37 °C and pH 8.0, using Tris-HCl as a buffer. The fluorescence spectroscopy results indicated that the binding of spermine to trypsin was a spontaneous binding process. The fluorescence of trypsin was quenched by spermine through the static quenching mechanism. By increasing the concentration of spermine, the thermal stability of trypsin was increased. The quantitative analysis of CD spectra revealed some information regarding the changes in the content of the α-helix and the β-sheet of the trypsin upon binding to spermine. It was also found that by increasing the concentration of spermine, the activity of 2 trypsin was increased. Molecular docking results also revealed the presence of one binding site with a negative value for the Gibbs free energy of the binding of spermine to trypsin. Further, the docking and fluorescence spectroscopy study revealed that van der Waals interactions and hydrogen bonds played a major role in stabilizing the complex. As a result, spermine could be considered as an activator and stabilizer for trypsin.

show abstract

Analysis of the interactions between Eudragit® L100 and porcine pancreatic trypsin by calorimetric techniques

Cited by 13 publications

References 27 publications

Protein–polyelectrolyte interactions

Protein–polyelectrolyte interactions

An Enteric-Coated Polyelectrolyte Nanocomplex Delivers Insulin in Rat Intestinal Instillations When Combined with a Permeation Enhancer

The effect of spermine on the structure, thermal stability and activity of bovine pancreatic trypsin

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