【作者】 杜健军；

【导师】 彭孝军；

【作者基本信息】 大连理工大学 ， 应用化学， 2009， 博士

【摘要】 生命活动是由许多生物活性物质参加的各种化学反应的结果,金属离子就是其中重要的一种。IIB族元素中,锌是生物体内必需的微量金属元素,直接参与机体内很多重要的生理过程,在维持蛋白质,核酸,肽类和激素等生物大分子的基本结构和实现其正常功能方面起着重要的作用。而IIB族元素中的镉和汞在较低浓度时就会对生物体产生较大的毒性和破坏性。因此,IIB族金属离子的研究和检测特别是对细胞内金属离子的成像检测具有重要的意义。与传统的金属离子检测方法相比,荧光探针检测因其兼备便捷、专一和灵敏的优点,并能对环境体系和生命体系中的分析对象进行实时原位检测而具有广泛的应用前景。因此设计合成金属离子荧光探针,在化学体系中试验其性质,结合荧光成像技术,检测细胞内某种金属离子的浓度变化及其分布。这将对探讨金属离子在生命活动中所发挥的作用以及对某种离子缺陷时出现的病理研究提供重要的帮助。因而用荧光探针检测金属离子成为了超分子化学以及生物医学等领域中重要的研究课题之一。本论文基于PCT原理设计合成了BODIPY类镉离子比率型荧光探针BCd1。在识别过程中,镉离子的配位作用减弱了共轭氮原子对荧光团的供电能力,从而引起了荧光波长近60 nm的蓝移,荧光量子产率增大约4倍。BCd1具有很强的抗干扰能力,同时具有较低的pKa（4.1）。BCd1可以完全区分Cd²⁺和Zn²⁺,并实现了在Hela细胞中镉离子的比率荧光成像。基于PET原理设计合成了一系列以BODIPY为荧光团,以开链硫醚作为识别基团的汞离子荧光探针BHg1-3。通过对醚链末端基团的修饰来调整探针分子的油/水溶性以及对汞离子的识别能力。研究表明,探针分子对目标离子的识别能力对探针分子的抗阴阳离子干扰以及细胞内汞离子成像具有重要的影响。设计合成了一系列基于汞离子促进水解反应的罗丹明类Hg²⁺荧光探针RHg1-3,并对该机理进行了详细阐述和验证。RHg1-3能够专一性地识别Hg²⁺,并具有很高的灵敏度。RHg1-3实现了在海水等水体系中ppb级Hg²⁺的检测,以及在Hela细胞内Hg²⁺的荧光显微成像。基于去质子和PCT原理设计合成了喹啉类锌离子比率荧光分子探针QZn1。在识别过程中,锌离子的配位作用导致荧光波长红移约90 nm,显示了典型的双波长比率荧光探针的光谱特征。更多还原

【Abstract】 The activity of life is an effect of different chemical reactions by lots of biological active substances,and metal ion is one important kind of them.Essential trace Zinc participates in many important physiological processes.For example,it plays critical roles in maintaining the structure and realizing the normal function of protein,nucleic acid,peptides,incretion and other big biology molecules.However,Cadmium and mercury are toxic and harmful to organism even at very low concentrations.So the detection of them especially the detection in vivo is important to the research of biology,medicine and environment science.Compared with tranditional methods for the detection of metal ions,the fluorescent probes have attracted much attention over the years for its many inherent merits including high specifity and high sensitivity.This technology has been widely used for the real-time detection of the molecular events both in the chemical and biological systems.We can design and synthesize metal ions fluorescent probes,and study the property of the probes in chemical system,and then detect the concentration changes and distribution of metal ions in cells on line according to confocal fluorescence imaging technology.This will be helpful to explore the function of metal ions in life activity and pathology of the shortness of metal ions and afford essential basis in the chemical and the biological systems.Therefore the BODIPY ratio-fluorescent Cd²⁺ probe BCd1 based on PCT mechanism has been designed and synthesized.The formation of complex BCdl/Cd2²⁺ weakened the electron-donating character,leading to a blue shift of 60 nm in fluorescence emission and an increase of quantum yield for about 4 fold.BCd1 shows strong anti-jamming and low pKa.It can distinguish Cd2²⁺ from Zn²⁺ and especially can be used in ratiometric fluorescence microscopy in Hela cells.The BODIPY fluorescent Hg²⁺ probe BHg1-3 based on PET mechanism have been designed and synthesized with a series of NS₂X₂ catenulate group as recogniation group.The coordination ability and oil/water solubility of the probes can be adjusted by different recognition moiety of catenulate NS₂X₂.The results indicate that comlexation ability between probes and ions is one of most important factors for interference and cell-imaging experiment.The rhodamine derivatives of Hg²⁺ fluorescent probes Rltg1-3 based on Hg²⁺-prompted hydrolysis mechanism have been designed and synthesized.The mechanism was studied and confirmed carefully.RHg1-3 exhibit Hg²⁺ selective among metal ions.RHg series have extremely highly sensitivity for Hg²⁺.RHg1-3 have enabled the detection of ppb level of Hg²⁺ in natural water samples like seawater.More importantly,they hava been also proved of Hg²⁺ fluorescence microscopy in Hela cells.The quinoline derivative of ratio-fluorescent Zn²⁺ probe QZn1 based on mechanism of PCT and deprotonation has been designed and synthesized.The formation of complex QZn1/Zn²⁺ leads to a 90 nm red shift in fluorescence emission,which shows typical ratio changes in spectra.更多还原