CAJ | 학술논문

目的:为了降低尿酸酶的免疫原性并提高其稳定性,利用PEG修饰荆对尿酸酶进行修饰,以期获得性质更优的治疗痛风的药物.方法:用相对分子质量为40kD,活化基团为羟基琥珀酰亚胺的PEG修饰剂对尿酸酶进行修饰,并比较修饰前后在酶解稳定性、pH稳定性及温度稳定性方面的差异,以及PEG修饰对体内半衰期,免疫原性的影响.结果:发现PEG修饰后尿酸酶的酶解稳定性显著提高,PEG化尿酸酶保留了原有尿酸酶80%的活性,体内半衰期从45min延长至696min.PEG化尿酸酶与抗体的结合能力为原型蛋白的1/8.体内的免疫原性也明显降低.结论:化学修饰后的尿酸酶可望成为潜在的治疗痛风的有效药物.
목적:위료강저뇨산매적면역원성병제고기은정성,이용PEG수식형대뇨산매진행수식,이기획득성질경우적치료통풍적약물.방법:용상대분자질량위40kD,활화기단위간기호박선아알적PEG수식제대뇨산매진행수식,병비교수식전후재매해은정성、pH은정성급온도은정성방면적차이,이급PEG수식대체내반쇠기,면역원성적영향.결과:발현PEG수식후뇨산매적매해은정성현저제고,PEG화뇨산매보류료원유뇨산매80%적활성,체내반쇠기종45min연장지696min.PEG화뇨산매여항체적결합능력위원형단백적1/8.체내적면역원성야명현강저.결론:화학수식후적뇨산매가망성위잠재적치료통풍적유효약물.
To modify uricase with PEG reagent in order to decrease uricase immunogenieity and increase its stability.Methods:The branched PEG of 40 kD was chosen to modify native uricase.The properties of the mod-ified uricase including the stabilities to protease,pH and temperature,in vivo half-life time,as well as the immu-nogenicity were evaluated.The pharmacokinetic profiles of the midofied uricase were studied in mice.Results:It is demonstrated that the conjugation of PEG to lysine residues of Candida utilis uricase resulted in higher tryp-sin resistance.reduced immune response.and prolonged in vivo half-life.PEG modified uricase retained 80% of the enzymatic activity of native uricase.In addition,it was found that half-life in serum of the intravenously injec-ted PEGylated uriease of up to 696 min was longer that that of native uficase of 45 min.Higher plasma drug con-centrations were also reached with dosing of the PEGylated uricase to mice.Furthermore,the binding affinity Was shown to be reduced for the PEG-uricase using ELISA assay.and it was one-eishth that of native uricase.Final-ly,it Was indicated that the PEG uficase induced a delayed immunoresponse in mice following repeated adminis-trations.Conclusion:These findings demonstrate that this chemically modified form of uricase may serve as a potentially effective drug to treat gout patients.