SIRT5调节剂的荧光筛选法研究  

Fluorescence-based screening for SIRT5 modulators

作　　者：李燕[1] 崔珺[2,3] 刘亭[4,3] 李勇军[2,3] 兰燕宇[2,3] 王爱民[2,3] 王永林[4,3] 何彬[2,3] 

机构地区：[1]贵阳医学院基础医学院,贵阳550004 [2]贵阳医学院民族药与中药开发应用教育部工程研究中心,贵阳550004 [3]贵阳医学院药学院,贵阳550004 [4]贵阳医学院贵州省药物制剂重点实验室,贵阳550004

出　　处：《中国新药杂志》2014年第15期1795-1800,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金(21302027);贵州省中药现代化科技产业研究开发专项(黔科合重G字[2013]4001);贵州省科技支撑计划社会发展攻关项目([2012]3079);贵州省高等学校创新能力提升计划(黔教合协同创新字[2013]04)

摘　　要：目的:鉴于高通量筛选方法是发现Sirtuin调节剂的重要手段,拟通过组蛋白去乙酰化酶SIRT5去琥珀酰化酶活的最新发现,开发出SIRT5酶活的"直接荧光测定法",并探讨其在筛选SIRT5特异型抑制剂方面的应用。方法:设计、合成了带有香豆素荧光基团的琥珀酰多肽底物,通过条件优化,我们建立了SIRT5调节剂的荧光筛选方法,再结合SIRT1/2/3活性的二次筛选,可以发现SIRT5的特异型调节剂。结果与结论:建立了SIRT5调节剂的荧光筛选方法,为最终SIRT5靶向治疗的药物发现奠定基础。Objective： The high-throughput screening is an efficient approach for discovering srtuin modulators. In the present study, we developed a fluorescence-based method for screening SIRT5 modulators based on SIRT5 novel enzymatic activity, the desuccinylase activity, and further explored its application in screening of SIRT5 specific inhibitors. Methods： We designed and synthesized a coumarin containing succinyl peptide as a fluorogenic substrate. By optimizing the conditions, we established a fluorescence-based method for screening SIRT5 modulators, which further combined with the secondary assay for screening SIRT1/2/3 inhibitors to find the specificity of SIRT5 inhibitors. Results and Conclusion： The fluorescence-based method for screening SIRT5 modulators will facilitate the drug discovery with SIRT5 as a therapeutic target

关 键 词：组蛋白去乙酰化酶 SIRTUIN 5 调节剂 荧光筛选方法 

分 类 号：R914.5[医药卫生—药物化学]