Research on PEGylated uricase by periodate oxidation and reductive- amination
10.11665/j.issn.1000-5048.20150318
- VernacularTitle:高碘酸氧化-还原胺化法对尿酸酶的聚乙二醇化研究
- Author:
Yangjian CHEN
1
;
Xinqi CHEN
;
Xiangyun ZHENG
;
Xiaoda SONG
Author Information
1. 浙江医药高等专科学校生物与制药学院；中国药科大学生命科学与技术学院
- Publication Type:Journal Article
- Keywords:
poly(ethylene glycol)derivative;
N-terminal site-specific modification;
uricase;
periodate oxidation;
reductive-amination
- From:
Journal of China Pharmaceutical University
2015;46(3):364-370
- CountryChina
- Language:Chinese
-
Abstract:
PEGylated uricase was prepared with the N-terminal amino site-specific modification by periodate oxidation followed by reductive-amination. A monomethoxy poly(ethylene glycol)intermediate was synthesized by amidation from monomethoxy poly(ethylene glycol)amine hydrochloride 20000(mPEG20000-NH2 ·HCl)with the relative molecular mass of 20 kD and N-(tert-butoxycarbonyl)-L-serine(Boc-Ser-OH), and then the Boc group of the intermediate was removed by trifluoroacetic acid(TFA)to produce the desired product Ser-mPEG20000. This compound could be oxidated by periodate to obtain a new poly(ethylene glycol)aldehyde derivative with high activity, which could be used to modify proteins with the N-terminal amino site-specific PEGylation after ultrafiltration, and the modification conditions to uricase by Ser-mPEG20000 were optimized. The structures of poly(ethylene glycol)intermediate and the target product were characterized by IR and 1H NMR, and the overall yield of the target product was 72. 8%. The preliminary modification to uricase indicated that the desired product Ser-mPEG20000 could modify proteins easily and efficiently. The optimal modification conditions of uricase PEGylated by Ser-mPEG20000 were obtained as follows: the molar ratio of Ser-mPEG20000 to uricase was 2 ∶1; the pH value of solution was 5. 0; the reaction temperature was 25 °C and the reaction time was 6 h.
