摘要
目的 用计算机虚拟筛选法寻找 BCR -ABL 酪氨酸激酶小分子抑制剂。方法 采用 ABL 酪氨酸激酶催化区在非活化时的独特构象作为受体结合位点,对有机小分子三维结构数据库进行 DOCK 筛选,寻找先导化合物;用M TT 法检测挑选的化合物对细胞增殖的抑制作用;用 W estern blot和免疫共沉淀方法检测化合物对 BCR -ABL 表达与活性的影响。结果 从 DOCK 筛选得到能量得分靠前的 1000个化合物中,选择了 15个化合物进行生物学测试。体外试验表明,8个化合物对 K 562细胞有明显的抑制作用 (IC 值在 10～200μm ol/L 之间),发现了两种新骨架结构 50的先导化合物,它们对 BCR -ABL 活性有明显的抑制作用,但对 ABL 的表达无影响。结论 筛选出对 BCR -ABL 酪氨酸激酶有明显抑制作用的两个先导化合物。计算机虚拟筛选方法通过对化合物数据库进行 “筛选”,可以大大提高先导化合物发现的效率,加快新药研究的步伐。
Objective To discover BCR-ABL tyrosine kinase inhibitors through structure based virtual screening. Methods Docking screening against the distinctive inactive conformation of the catalytic domain of BCR-ABL tyrosine kinase was performed on 3D database. The MTT assay was performed to assess the viability of the tumor cells treated with selected compounds. The amount and kinase activity of BCR-ABL protein were detected in the presence of compounds by Western blot analysis and immunoprecipitation. Results From the top 1 000 compounds with the best DOCK energy score, 15 compounds were selected for biological assay. Eight out of 15 compounds showed notable inhibitory activity against Ph+ human K562 cells with IC50 values ranging from 10 to 200 μmol/L. In cell-based assays of ABL tyrosine phosphorylation, the ability of two kinds of novel, structurally diverse, lead compounds to inhibit ABL kinase activity was observed. However, no significant differences in the amount of BCR-ABL protein were noted on the ABL immunoblot in the presence of these lead compounds. Conclusions Two promising lead compounds were discovered to inhibit BCR-ABL tyrosine kinase activity. Virtual screening technique has been proven to narrow down the size of screening compound libraries to the most prospective drug candidates with high success rates.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2004年第2期145-149,i004,共6页
Acta Academiae Medicinae Sinicae
基金
国家高技术研究发展计划项目(863项目)(2001AA234021-9)资助~~
