Thiolated pectins: In vitro and ex vivo evaluation of three generations of thiomers

• Published in: Acta biomaterialia 2021-11, Vol.135, p.139-149
• Main Authors: Knoll, Patrick, Le, Nguyet-Minh Nguyen, Wibel, Richard, Baus, Randi Angela, Kali, Gergely, Asim, Mulazim Hussain, Bernkop-Schnürch, Andreas
• Format: Article
• Language: English
• Subjects: 3rd generation thiomer; Colorimetry; Cysteine; Cytotoxicity; Diffusion; Glutathione; Interpenetration; L-glutathione; Ligands; Mucoadhesion; Mucosa; Mucus; Mucus diffusion; NMR; Nuclear magnetic resonance; Pectin; Penetration; Polymers; S-protection; Swelling; Thiol/disulfide exchange; Thiolated polymers; Toxicity; Viscosity
• ISSN: 1742-7061; 1878-7568
• DOI: 10.1016/j.actbio.2021.08.016

In recent decades, three generations of thiomers have been developed with the main purpose of obtaining enhanced interactions with mucosal tissues. Therefore, many different types of thiolated ligands have been generated and attached to polymeric backbones. The aim of this study was to synthesize all three generations of thiomers and to directly compare their properties regarding mucus penetration and mucoadhesion. Starting from pectin, the unprotected thiomer pectin-cysteine (Pec-Cys), the preactivated S-protected thiomer pectin-cysteine-mercaptonicotinic acid (Pec-Cys-MNA) and the less reactive S-protected thiomer pectin-cysteine-glutathione (Pec-Cys-GSH) were synthesized and characterised by FT-IR, NMR, and colorimetric studies. The polymers were evaluated regarding their cytotoxicity, swelling behaviour, viscosity after mixing with mucus, mucus diffusion, penetration into mucosa, and mucoadhesion. The amount of the three ligands (Cys, Cys-MNA and Cys-GSH) bound to the polymer was determined to be in the range of 193-196 µmol/g. All polymers showed no cytotoxicity. Viscosity of the mixture of Pec-Cys-MNA and Pec-Cys-GSH with mucus increased 21.5- and 26.7-fold, respectively, compared to the unmodified polymer within 3 hours. Swelling, mucoadhesion, interpenetration and mucus diffusion were increased in the following rank order: Pec-Cys < Pec-Cys-MNA < Pec-Cys-GSH. Results of mucoadhesion study indicated a 7.4 and 8.1-fold increase of Pec-Cys-MNA and Pec-Cys-GSH, respectively, compared to the unmodified polymer. As the less reactive S-protected thiomer exhibited higher mucoadhesive properties than the other thiomers, this study provides evidence for the superior mucoadhesion of 3rd generation thiomers. Three generations of thiolated polymers have been developed bearing different types of thiol ligands with the main purpose of enhancing mucus interactions. In this study, all generations were synthesized on the polymeric backbone of pectin for the first time to directly compare their mucus penetrating and mucoadhesive properties. 1st generation exhibited covalently bound L-cysteine moieties. For 2nd generation, thiols of cysteines were S-protected with 2-mercaptonicotinic acid (MNA), resulting in high reactive disulfide bonds. 3rd generation was synthesized by a thiol/disulfide exchange of glutathione with MNA, producing a less reactive disulfide bond. Mucus penetrating and mucoadhesive properties were found to be increased as follows: 1st generation < 2nd