新型抗HBV化合物:-βL-D4A对DNA聚合酶δ的作用及其机制  

Effect of β-L-D4A on DNA polymerase δ

作　　者：李岩[1] 林菊生[1] 张颖慧[1] 王晓燕[1] 何星星[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝病研究所,湖北武汉430030

出　　处：《胃肠病学和肝病学杂志》2008年第4期318-321,324,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金重点项目(30330680)

摘　　要：目的对DNA聚合酶δ进行纯化和鉴定,通过检测β-L-D4A对DNA聚合酶δ的作用,观察β-L-D4A的毒副作用。方法运用多次离子交换层析的方法提取、纯化人Hela细胞的DNA聚合酶δ;检测β-L-D4A对DNA聚合酶δ的酶动力学作用。结果提纯并鉴定了DNA聚合酶δ,收得率是15%,比活力是1 911 ukat/mg。β-L-D4A为抑制剂时,Km是2.45μmol/L,IC50是150.1μmol/L,K i是132.7μmol/L,均提示β-L-D4A对DNA聚合酶δ的抑制作用远小于拉米夫定。结论β-L-D4A是DNA聚合酶δ的竞争性抑制剂,它对该酶的毒副作用远小于拉米夫定,可望成为更高效低毒的抗HBV类药物。Objective To purify and characterize DNA polymerase δ and investigate the side effect of β-L-D4A on DNA polymerase δ. Methods Ion exchange chromatography was used to separate DNA polymerase δ from crude extract of human Hela cell. Detailed kinetic parameters were determined for β-L-D4A against DNA polymerase δ. Results DNA polymerase δ was purified and characterized with 15% yield and 1 911 ukat/mg specific activity. The kinetic constants of β-L-D4A against DNA polymerase δ were Km 2.45 μmol/L, IC50 150.1μmol/L, Ki 132.7 μmol/L,which confirmed that the inhibition of β-L-D4A was less than lamivudine. Conclusion These findings predict that β-L-D4A may be a more effective anti-virus drug.

关 键 词：核苷类 -βL-D4A DNA聚合酶Δ 酶动力学 毒副作用 

分 类 号：R979.5[医药卫生—药品]