title: Mucin-based stationary phases as tool for the characterization of drug-mucus interaction.
abstract: Mucin glycoproteins belong to a class of high molecular weight, heavy glycosylated, proteins that together with water, salts and lipids constitute mucous secretions. Particular disease states (e.g. obstructive chronic bronchitis and ovarian tumor) are known to modify the composition and the thickness of those barriers. Therefore, it is important to address whether the absorption of potential drug candidates to be administered is influenced by the presence of interaction with this class of proteins. Typically, the methods adopted to characterize drug-protein interaction are dialysis, ultrafiltration and gel filtration. Besides these, bio-affinity chromatographic methods have demonstrated to be valuable tools offering the advantageous characteristics such as simplicity, efficiency, high-throughput capability and robustness. The present contribution reports on the synthesis and analytical characterization of a new chromatographic stationary phase based on covalently immobilized mucin and explores the use of LC-UV affinity zonal chromatography as a tool to screen drugs for their affinity to mucin. A series of different binding chemistries for the covalent linkage of mucin to silica-based supports as well as distinct immobilization protocols (static and dynamic) have been evaluated in order to optimize surface coverage. Resultant stationary phases have been characterized chromatographically by studying the effect of mobile phase and analyte structure on the distribution and retention of test compounds. As conclusive study, we report the evaluation of the retention characteristics of 41 drug-like compounds (having heterogeneous chemical properties) for their interaction with this novel stationary phase.
authors.affiliation: University of Vienna, Department of Analytical Chemistry, Währinger Strasse 38, 1090 Vienna, Austria; TI-COAST, Van 't Hoff Institute for Molecular Sciences, Science Park 904, 1098 XH Amsterdam, Netherlands. Electronic address: a.gargano@uva.nl.
authors.forename: Andrea F G
authors.initials: AF
authors.lastname: Gargano
authors.affiliation: University of Tuebingen, Institute of Pharmaceutical Sciences, Auf der Morgenstelle 8, 72076 Tuebingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lämmerhofer
authors.affiliation: R&D iMed RIA, Astra Zeneca, Pepparedsleden 1, Mölndal SE-43183, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Lönn
authors.affiliation: TI-COAST, Van 't Hoff Institute for Molecular Sciences, Science Park 904, 1098 XH Amsterdam, Netherlands.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Schoenmakers
authors.affiliation: R&D iMed RIA, Astra Zeneca, Pepparedsleden 1, Mölndal SE-43183, Sweden.
authors.forename: Tomas
authors.initials: T
authors.lastname: Leek
mesh_terms: D000042:Absorption; D000818:Animals; D002846:Chromatography, Affinity; D004347:Drug Interactions; D009077:Mucins / Q000737:chemistry* / Q000378:metabolism; D009093:Mucus / Q000737:chemistry / Q000378:metabolism*; D012822:Silicon Dioxide / Q000737:chemistry; D013552:Swine
chemical_list: D009077:Mucins; D012822:Silicon Dioxide
keywords: Affinity chromatography; Drug–protein affinity; Liquid chromatography; Mucin; Protein stationary phases