title: Monitoring the in vitro enzyme-mediated degradation of degradable poly(ester amide) for controlled drug delivery by LC-ToF-MS.
abstract: To scrutinize materials for specific biomedical applications, we need sensitive and selective analytical methods that can give more insight into the process of their biodegradation. In the present study, the enzymatic degradation of multiblock poly(ester amide) based on natural amino acids, such as lysine and leucine, was performed with serine proteases (α-chymotrypsin (α-CT) and proteinase K (PK)) in phosphate-buffered saline solution at 37 °C for 4 weeks. Fully and partially degraded water-soluble products were analyzed by liquid chromatography hyphenated with time-of-flight mass spectrometry using an electrospray interface (LC-ESI-ToF-MS). Tracking the release of monomeric and oligomeric products into the enzyme media during the course of enzymatic degradation revealed the preferences of α-CT and PK toward ester and amide bonds: both α-CT and PK showed esterase and amidase activity. Although within the experimental time frame up to 30 and 15% weight loss was observed in case of α-CT and PK, respectively, analysis by size exclusion chromatography showed no change in the characteristic molecular-weight averages of the remaining polymer. This suggests that the enzymatic degradation occurs at the surface of this biomaterial. A sustained and linear degradation over a period of 4 weeks supports the potential of this class of poly(ester amide)s for drug delivery applications.
authors.affiliation: Analytical Chemistry Group, Van't Hoff Institute for Molecular Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands. a.ghaffar@uva.nl
authors.forename: A
authors.initials: A
authors.lastname: Ghaffar
authors.forename: G J J
authors.initials: GJ
authors.lastname: Draaisma
authors.forename: G
authors.initials: G
authors.lastname: Mihov
authors.forename: A A
authors.initials: AA
authors.lastname: Dias
authors.forename: P J
authors.initials: PJ
authors.lastname: Schoenmakers
authors.forename: Sj
authors.initials: S
authors.lastname: van der Wal
mesh_terms: D000577:Amides / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002417:Cattle; D002853:Chromatography, Liquid; D002918:Chymotrypsin / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000379:methods; D019286:Endopeptidase K / Q000378:metabolism*; D005658:Fungi; D007930:Leucine / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008970:Molecular Weight; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000577:Amides; D001672:Biocompatible Materials; D011091:Polyesters; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D019286:Endopeptidase K; D007930:Leucine; D008239:Lysine