基于luciferin-luciferase体系的肼生物发光探针的设计、合成与光学评价  

Design,synthesis and optical evaluation of a hydrazine bioluminescent probe based on luciferin-luciferase system

作　　者：康婷 海澳 马雅茹 柯博文[3] 秦勇[1] 刘小宇[1] 仇丽颖 KANG Ting;HAI Ao;MA Yaru;KE Bowen;QIN Yong;LIU Xiaoyu;QIU Liying(Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry,West China School of Pharmacy,Sichuan University,Chengdu,Sichuan,610041 P. R. China;School of Pharmacy,Southwest Minzu University,Chengdu,Sichuan,610041 P. R. China;Laboratory of Anaesthesiology & Critical Care Medicine,West China Brain Research Center,West China Hospital,Sichuan University,Chengdu,Sichuan ,610041 P. R. China)

机构地区：[1]四川大学华西药学院教育部药物靶向与给药系统重点实验室,四川成都610041 [2]西南民族大学药学院,四川成都610041 [3]四川大学华西医院转化神经科学中心麻醉与危重急救研究室,四川成都610041

出　　处：《华西药学杂志》2019年第4期315-319,共5页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:81773685)

摘　　要：目的设计并合成一个用于肼(N_2H_4)可视化分析检测的小分子生物发光探针(bioluminescence probe for hydrazine,BLHy),并评价其光学响应性能。方法在光学报告的基团羟基萤光素(D-luciferin)上适当修饰位点,通过合理设计,引入肼的特异性识别基团(4-溴丁酸酯),设计和构建一个小分子生物发光探针BLHy。通过探针与底物反应后的信号强度和光学波长的变化,研究探针在肼可视化检测和分析中的应用。结果成功设计并合成了一个小分子生物发光探针BLHy。该探针在Tris-HCl缓冲体系中能特异性对肼进行响应,强度是其他物质的4~17倍,表现出良好的选择性;探针对0~333μmol·L^-1肼的响应具有良好的浓度依赖性(r^2=0. 9987)。结论首次以luciferin-luciferase生物发光体系作为分析技术,设计并合成了用于肼检测的生物发光探针BLHy,为进一步研究肼在复杂自然环境和生物体系中的作用和机制提供了有力的工具。OBJECTIVE To design, synthesize and evaluate a highly sensitive and selective bioluminescence probe ( BLHy) for the visable detection of hydrazine in vitro. METHODS A small molecular bioluminescence probe BLHy was designed and constructed by introducing the specific identification group (4 - bromobutyrate) of hydrazine on the appropriate site of the optical report group D-luciferin with a rational design strategy. The application of the probe in visable detection and analysis of hydrazine was studied by the changes of optical signals and wavelength after the reaction between the probe and the target substrate. The capability of this probe for tracking N2H4 was carefully assessed in vitro by using the Amersham Imager 600. RESULTS A small molecular bioluminescence probe BLHy was successfully designed and synthesized. The specific response intensity of BLHy to the hydrazine in Tris . HC1 buffer was 4-17 times that of other substances, indicating a good selectivity. In addition, BLHy showed an excellent linear relationship between its responsiveness and the concentrations of hydrazine at the range of 0 - 333 pjnol . L^-1 ( r^2 = 0. 9987 ). CONCLUSION The hydrazine probe BLHy, developed on the basis of the bioluminescence system for the first time, had been successfully employed for specific detection of hydrazine in vitro. As a result, this probe represented a powerful tool for investigating the role and underlying mechanism of hydrazine in the complicated environmental and biological systems.

关 键 词：肼 BLHy 羟基萤光素 荧光素酶 探针 Luciferin-luciferase体系 生物发光 4-溴丁酸酯 

分 类 号：R914[医药卫生—药物化学]