title: Purified PEGylated porcine glucagon-like peptide-2 reduces the severity of colonic injury in a murine model of experimental colitis.
abstract: The rapid degradation of porcine glucagon-like peptide-2 (pGLP-2) by the enzyme dipeptidyl peptidase-IV (DPP-IV) is the main impediment in the development of pGLP-2 as a potential therapeutic agent for intestinal dysfunction and damage. In this study, one mono-modified Lys(30)-polyethylene glycol (PEG)-pGLP-2 was prepared using mPEG-succinimidyl propionate. To determine the optimized condition for PEGylation, the reactions were monitored by RP-HPLC and MALDI-TOF-MS. Stability was tested in purified DPP-IV in vitro. In vivo, the protective effects for colonic injury were measured in dextran sulfate sodium (DSS)-induced colitis in mice. The monoPEGylated products reached the maximum yield at 4:1 ratio of mPEG5k-SPA to pGLP-2. An effective method of successfully separating PEGylated pGLP-2 from mPEG-SPA5kD using CM Sepharose Fast Flow resin was established. The half-life of Lys(30)-PEG-pGLP-2 was 16-fold longer than that of pGLP-2 in DPP-IV. The DSS mice exhibited marked weight loss), which was significantly reduced by Lys(30)-PEG-pGLP-2 therapy. DSS treatment significantly increased colonic damage score, which was significantly reduced by administration of Lys(30)-PEG-pGLP-2 in DSS-mice. DSS-induced colitis clearly induced Myeloperoxidase activity in the colon, which was significantly reduced by treatments with 3% DSS-pGLP-2 or 3% DSS-PEG-pGLP-2. These results showed that site-specific Lys(30)-PEG-GLP-2 was resistant to degradation and reduced the severity of colonic injury in murine colitis. The enhanced biological potency of this product highlighted its potential as a therapeutic agent for intestinal diseases.
authors.affiliation: Institute of Animal Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Ke-ke
authors.initials: KK
authors.lastname: Qi
authors.affiliation: Institute of Animal Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
authors.affiliation: Institute of Animal Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wan
authors.affiliation: Institute of Animal Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Xiao-ming
authors.initials: XM
authors.lastname: Men
authors.affiliation: Institute of Animal Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China. Electronic address: zjsnkyxzw@163.com.
authors.forename: Zi-wei
authors.initials: ZW
authors.lastname: Xu
mesh_terms: D000818:Animals; D002908:Chronic Disease; D003092:Colitis / Q000139:chemically induced / Q000188:drug therapy* / Q000401:mortality / Q000473:pathology; D003106:Colon / Q000378:metabolism* / Q000473:pathology; D016264:Dextran Sulfate / Q000633:toxicity; D004195:Disease Models, Animal; D053767:Glucagon-Like Peptide 2 / Q000737:chemistry / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology*; D013552:Swine
chemical_list: D053767:Glucagon-Like Peptide 2; D011092:Polyethylene Glycols; D016264:Dextran Sulfate
keywords: Colitis; Dextran sulfate sodium; Ion-exchange chromatography; PEGylation; Porcine glucagon-like peptide-2