新型核苷类似物β-L-D4A对人DNA聚合酶β和δ的作用及其机制  

Effect ofβ-L-D4A on human DNA polymerasesβandδand its mechanism

作　　者：李岩[1] 林菊生[1] 张颖慧[1] 王晓燕[1] 何星星[1] 汪鸿[1] 高玲玲[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,湖北省武汉市430030

出　　处：《世界华人消化杂志》2008年第6期621-628,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金重点;No.30330680~~

摘　　要：目的:对人DNA聚合酶δ进行纯化和鉴定,通过检测β-L-D4A对人DNA聚合酶β和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对人DNA聚合酶β和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1911 Ukat/mg.β-L-D4A为抑制剂时,对人DNA聚合酶β和δ的酶动力学参数分别是Km=1.86 umol/L,2.45 umol/L,IC50 =25.21μmol/L和150.1μmol/L,Ki=24.03μmol/L和132.7μmol/L,结果均提示β-L-D4A对DNA聚合酶β和δ的抑制作用远小于拉米夫定.结论:p-L-D4A是人DNA聚合酶β和δ的竞争性抑制剂,他对这两种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.AIM：To purify and characterize the DNA polymerase δ and investigate the side effects of β-L-D4A（a novel nucleoside analog）on human DNA polymerases β and δ. METHODS：Human DNA polymerase δ was separated from crude extract of human Hela cells by ion exchange chromatography.Detailed kinetic parameters were determined for β-L- D4A against DNA polymerases β and δ. RESULTS：Human DNA polymerase δ was purified and characterized with a yield of 15% and a specific activity of 1911 Ukat/mg.When β-L-D4A was used as an inhibitor,the kinetic parameters of human DNA polymerases β and were Km=1.86μmol/L and 2.45μmol/L,IC50 =25.21μmol/L and 150.1μmol/L,Ki=24.03μmol/L and 132.7μmol/L,respectively,suggesting that the inhibitory effect of β-L-D4A on human DNA polyrnerases β and δ was less than that of lamivudine. CONCLUSION：β-L-D4A is a competitive inhibitor of DNA polymerases β and δ.Its toxic side effects on DNA polymerases β and δ are less than lamivudine.It can be expected to become a high efficient and low toxic anti-HBV agent.

关 键 词：核苷类 Β-L-D4A 人DNA聚合酶β和δ 酶动力 毒副作用 

分 类 号：R96[医药卫生—药理学]