title: The potential of cystine-knot microproteins as novel pharmacophoric scaffolds in oral peptide drug delivery.
abstract: Within this study, the potential of three clinically relevant microproteins (SE-AG-AZ, SE-EM and SE-EP) with cystine-knot architecture as pharmacophoric scaffolds for oral peptide delivery was investigated. Cystine-knot microproteins (CKM) were analysed regarding their stability towards the most important gastrointestinal secreted and membrane bound proteases in physiological concentrations. In addition, their permeation behaviour through freshly excised rat intestinal mucosa as well as important parameters such as aggregation behaviour, stability in rat plasma and isoelectric point were evaluated and compared to the properties of the model peptide drugs bacitracin and insulin. Aggregation studies indicate that under physiological conditions between 25 and 70% of the CKMs occur as monomers, whereas the rest forms di- and trimers. Pepsin and elastase cause no or only minor degradation to CKMs, whereas trypsin and chymotrypsin degrade CKMs extensively. Removing the theoretical chymotrypsin cleavage site from a CKM, however, led to stabilization towards this protease. Two of the three evaluated CKMs are stable against membrane bound proteases. P(app) values were determined to be 5.96 +/- 0.98 x 10(-6) and 6.63 +/- 0.47 x 10(-6) cm/s. In conclusion, this study indicates that CKM are promising novel pharmacophoric scaffolds for oral peptide delivery.
authors.affiliation: ThioMatrix GmbH, Research Center Innsbruck, Mitterweg 24, 6020 Innsbruck, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Werle
authors.forename: Thierry
authors.initials: T
authors.lastname: Schmitz
authors.forename: Hong-Lei
authors.initials: HL
authors.lastname: Huang
authors.forename: Alexander
authors.initials: A
authors.lastname: Wentzel
authors.forename: Harald
authors.initials: H
authors.lastname: Kolmar
authors.forename: Andreas
authors.initials: A
authors.lastname: Bernkop-Schnürch
mesh_terms: D000284:Administration, Oral; D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000737:chemistry*; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D010455:Peptides / Q000008:administration & dosage* / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D051381:Rats
chemical_list: D010455:Peptides; D011506:Proteins; D003553:Cystine