基于氧杂蒽的近红外活性硫比率荧光探针的合成和性能研究

The abnormal levels of reactive sulfur species in the organism are associated with many human diseases. Accordingly, much effort has been given to the development of reactive sulfur quantitative detection method with good selectivity and high sensitivity. A detailed understanding the relationship between the produce, distribution and the level of fluctuation of these reactive sulfur species with cell activity, body physiology and pathology have important scientific significance. This project aims to design and synthesis of novel near infrared xanthene dyes, build general synthetic method of near infrared TBET-based xanthene ratio fluorophore group. The project will also develop a series of novel near infrared xanthene fluorescent ratio probes for the measurement of single reactive sulfur species by introduction of vinyl phosphate, phosphoryl, aryl phosphine and other auxiliary function group for reactive sulfur identification. The item will address the problems of the current fluorescent probe such as poor stability and biological compatibility, difficult to be quantitative. Systematic studies on the influence of different functional groups and functional groups position to the optical physical properties, the photochemical properties, selectivity and biological compatibility of the probe will be carried out. The project use modern advanced testing and characterization methods, to realize real time distribution and concentration quantitative detection of reactive sulfur species in biological systems, provide powerful tools and reliable methods for the further study of distribution in organism and role in the process of life of reactive sulfur.

生物体内活性硫物种水平异常能引起多种疾病，因此发展选择性好、灵敏度高的活性硫定量检测方法，详尽认识这些活性硫化合物的产生、分布及水平波动对细胞活性、机体生理和病理等研究有着重要的科学意义。本项目拟设计合成结构新颖的近红外氧杂蒽类荧光团，建立基于跨键能量转移的近红外氧杂蒽比率荧光团通用合成方法，引入烯基磷酸酯、磷酰基、芳基膦等活性硫识别辅助功能团，开发系列新型近红外氧杂蒽比率测量单一活性硫物种的荧光探针，努力解决目前荧光探针分子稳定性差、生物兼容性不佳、无法准确定量等问题。系统地研究不同功能团和功能团位置对探针的光物理性质、光化学性质、选择性、生物兼容性等的影响。利用现代先进的测试手段和表征方法，实现对活性硫物种在生物体系中实时分布及浓度的定量检测，为进一步研究活性硫在生物体中的分布、生命过程中的作用提供有力工具和可靠手段。

生物体内存在多种重要的小分子活性硫物种，它们的浓度不尽相同，所起的生理功能各异，且缺一不可。这些活性硫水平异常与多种疾病密切相关，因此发展选择性好的活性硫荧光探针对认识活性物种与某些疾病的关联具有重要科学意义。项目组设计合成了一系列荧光探针（主要是活性硫探针），运用紫外、荧光和共聚焦成像研究了它们的性能及在成像方面的应用。研究发现苯并吡喃盐既可作为亚硫酸响应基团，也可作为线粒体定位基，基于该基团，项目组发展了一例能在细胞和动物水平实时监测Cys代谢的荧光探针，成果作为封面文章发表；内质网中活性硫物种Cys，GSH和活性氧HOCl是彼此相关的，项目组合成了首例连续响应双光子荧光探针，发现细胞内存在还原应激；报道了第一例次氯酸可逆探针，文章发表后成高被引论文；项目组还发明了一个基于罗萨明的硫化氢探针和两个双功能探针的便捷合成方法，并申报专利3项，这三个探针成像效果好，具有较强应用潜力。项目研究成果获得河南省教育厅科技成果一等奖1项，发表影响因子3以上的SCI论文18篇，中科院1区论文11篇，高被引论文1篇，培养毕业硕士研究生13名。