title: Nanosized insulin-complexes based on biodegradable amine-modified graft polyesters poly[vinyl-3-(diethylamino)-propylcarbamate-co-(vinyl acetate)-co-(vinyl alcohol)]-graft-poly(L-lactic acid): protection against enzymatic degradation, interaction with Caco-2 cell monolayers, peptide transport and cytotoxicity.
abstract: Non-parenteral insulin delivery by the oral route is limited by epithelial barriers and enzymatic degradation. Nanosized insulin-complexes based on amine modified comb-like polyesters were designed to overcome these barriers. Protection of insulin in nanocomplexes (NC) from enzymatic degradation was investigated. The interaction with enterocyte-like Caco-2 cells in terms of cytotoxicity, transport through and uptake in the cell layers was evaluated by measuring transepithelial electrical resistance (TEER), release of lactate dehydrogenase (LDH) and insulin transport. The protection capacity of NC and their interaction with Caco-2 cells varied strongly as a function of lactide-grafting (hydrophobicity). With increasing lactide-grafting (P(26) < or = P(26)-1(LL) < or = P(26)-2(LL)) NC protected up to 95% of the insulin against degradation by trypsin. Transport and uptake into cell monolayers increased with higher l-lactid grafting. About 25% of a 1.25mg/ml TRITC-insulin NC dispersion with P(26)-2(LL) was recovered in Caco-2 cells after 90 min. A concentration dependent cytotoxicity profile was observed showing elevated LDH release and decreased TEER values. The cytotoxicity correlates with the surfactant like character of the polymers, decreasing the surface tension to 46 mN/m for the amphiphilic P(26)-2(LL). The observed TEER decrease was reversible after 20 h, suggesting that the biodegradable comb-polyesters offer a promising approach to overcome mucosal barriers.
authors.affiliation: Department of Pharmaceutics and Biopharmacy, Philipps-Universität, Marburg, Germany. pepsi51@gmx.de
authors.forename: Michael
authors.initials: M
authors.lastname: Simon
authors.forename: Isabel
authors.initials: I
authors.lastname: Behrens
authors.forename: Lea Anne
authors.initials: LA
authors.lastname: Dailey
authors.forename: Matthias
authors.initials: M
authors.lastname: Wittmar
authors.forename: Thomas
authors.initials: T
authors.lastname: Kissel
mesh_terms: D018938:Caco-2 Cells; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D004339:Drug Compounding; D004355:Drug Stability; D017097:Electric Impedance; D006801:Humans; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000378:metabolism*; D007328:Insulin / Q000737:chemistry / Q000378:metabolism*; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D053758:Nanoparticles; D011091:Polyesters / Q000138:chemical synthesis / Q000737:chemistry* / Q000633:toxicity; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
chemical_list: D004337:Drug Carriers; D007004:Hypoglycemic Agents; D007328:Insulin; D011091:Polyesters; D007770:L-Lactate Dehydrogenase; D014357:Trypsin