Pitarresi Giovanna, Palumbo Fabio Salvatore, Triolo Daniela, Fiorica Calogero, Giammona Gaetano
a Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche, Sezione di Chimica e Tecnologie Farmaceutiche , Università degli Studi di Palermo , Palermo , Italy.
b Institute of Biophysics at Palermo , Italian National Research Council , Palermo , Italy.
Drug Dev Ind Pharm. 2017 Sep;43(9):1480-1488. doi: 10.1080/03639045.2017.1319380. Epub 2017 Apr 27.
Aim of this work was the synthesis of a methacrylic hyaluronic acid (HA) derivative and the production, via photocrosslinking, of related hydrogels loaded with an endopeptidase intended for a potential oral treatment of celiac disease.
The methacrylic derivative of HA was prepared through a one-pot procedure involving the reaction with ethylenediamine (EDA) and methacrylic anhydride (MA). The obtained derivative, named HA-EDA-MA, was used to prepare photocrosslinked hydrogels loaded with a prolyl endopeptidase derived from Flavobacterium meningosepticum (PEP FM) able to detoxify gliadin. Obtained hydrogels were recovered as gels or freeze-dried powders.
Hydrogels obtained as freeze-dried powders, are able to protect loaded enzyme from degradation due to freeze-drying process and from alteration during storage, overall in the presence of a cryoprotectant. All photocrosslinked HA-EDA-MA hydrogels (gels and powders) release PEP FM in simulated intestinal fluid in sustained manner and in active form. HA-EDA-MA hydrogels are nontoxic as demonstrated through in vitro studies on BALB 3T3 cells.
Prepared hydrogels show a potential application for oral treatment of celiac disease thanks to the possibility to release enzymes able to detoxify the gliadin peptide that induces the immunogenic response.
本研究旨在合成一种甲基丙烯酸透明质酸(HA)衍生物,并通过光交联制备负载内肽酶的相关水凝胶,用于乳糜泻的潜在口服治疗。
通过一锅法制备HA的甲基丙烯酸衍生物,该方法涉及与乙二胺(EDA)和甲基丙烯酸酐(MA)的反应。将所得衍生物命名为HA-EDA-MA,用于制备负载源自脑膜败血黄杆菌的脯氨酰内肽酶(PEP FM)的光交联水凝胶,该酶能够使麦醇溶蛋白解毒。所得水凝胶以凝胶或冻干粉末形式回收。
以冻干粉末形式获得的水凝胶能够保护负载的酶免受冻干过程的降解以及储存期间的变化,总体上是在存在冷冻保护剂的情况下。所有光交联的HA-EDA-MA水凝胶(凝胶和粉末)在模拟肠液中以持续方式并以活性形式释放PEP FM。通过对BALB 3T3细胞的体外研究表明,HA-EDA-MA水凝胶无毒。
制备的水凝胶显示出在乳糜泻口服治疗中的潜在应用,这得益于其能够释放能够使诱导免疫原性反应的麦醇溶蛋白肽解毒的酶。