【作者】 何广杰；

【导师】 段春迎； 何成；

【作者基本信息】 大连理工大学 ， 无机化学， 2010， 博士

【摘要】 分子和离子识别是化学、生物学和材料科学中一个至关重要的现象,是超分子化学研究的基础。分子识别发生在一个为人们不能直接感知的微观世界里,因此想了解和研究分子间的识别行为,需要通过具有特殊性质的分子探测器件输出可观测的宏观信号。在这些信号中,最为直接的方式是以光信号的形式表达出来,即可视化。将经过特殊设计的荧光分子探针引入到待测体系中,能够将分子识别的信息转换成荧光信号传递给外界,从而使人与分子间的对话成为可能,架起宏观世界和微观世界联系的桥梁,是当前分子离子探针研究中的重点和热点。本论文包含以下几方面研究内容：1.活体细胞中Cu2+的PET荧光探针：顺磁性的Cu2+对荧光分子激发态的淬灭性质,使得高选择性和灵敏度的荧光增强型的Cu2+荧光探针的设计成为一项富有挑战性的工作。本论文设计合成了三个Cu2+荧光探针。探针L1(2-羟基-1-萘醛和联苯二腙的双臂化合物)以荧光淬灭的方式选择性地识别Cu2+,但Hg2+有干扰；探针L2(丁基萘酐肼和叔丁基苯酚二醛的双臂化合物)通过荧光淬灭高选择性地识别Cu2+,Hg2+对其无干扰,并且能够应用于活体细胞荧光成像。在此探针分子研究的基础上,通过电化学测量和量子化学理论计算,精心选择了与Cu2+的氧化还原电位相匹配的电子给体和受体组分,合成了具有合适前线轨道能级的探针分子L3(7-N,N-二乙基氨基香豆醛和联苯二腙的双臂化合物)。L3本身荧光极其微弱,和Cu2+配位后,由于轨道能级的合理匹配,其分子内的光诱导电子转移过程(PET)被阻断,荧光显著增加75倍。L3荧光探针具有高选择性、高灵敏度、良好的水溶性等优点。L3和Cu2+的活体细胞共聚焦荧光成像表明L3是一个非常优良的可用于生命体系的Cu2+荧光探针。2.FRET的比率荧光探针和全光谱荧光分子的设计合成：利用能量匹配的荧光基团之间的荧光共振能量转移(FRET)设计合成具有多个荧光通道的分子是目前荧光探针和荧光材料研究中的热点。本论文中的探针分子RC1和RC2分别由罗丹明6G(绿色荧光)或罗丹明B(红色荧光)与7-N,N-二乙基氨基香豆酰氯(蓝色荧光)荧光团通过乙二胺桥联而成,选择性和竞争性实验表明这两个荧光探针能通过FRET以比率荧光选择性地识别Cu2+离子。CRB和CR6G由罗丹明B或者罗丹明6G与7-N,N-二乙基氨基香豆醛通过肼基桥联而成,这两个荧光探针以紫外可见光谱和荧光光谱双通道同时识别水溶液中的Cu2+和Hg2+。在这些双荧光基团分子探针的研究基础上,本论文进一步将具有锐线发射光谱的稀土离子引入,合成具有三荧光通道的荧光材料：单分子荧光材料材料RC1-Eu, RC1-Eu由RC1(包含蓝色和绿色荧光基团)和Eu(发射红色荧光)通过配位反应合成。通过控制其中的能量转移过程和效率,RC1-Eu能分别发射蓝、绿、红三原色荧光,并且在合适的激发波长处同时发射蓝、绿、红三原色荧光而形成白光。3.基于金属配合物的生物小分子荧光探针的设计合成：单糖、氨基酸、核糖核酸等生物小分子荧光探针在生命科学、医学等方面具有重要应用价值。在纯有机荧光探针分子的研究基础上,本论文设计合成了一系列具有荧光活性的铱、铕、钌等金属配合物的生物小分子荧光探针。探针Ir-OH(邻菲啰啉苯硼酸的铱配合物)在各种单糖中以荧光淬灭的方式选择性地识别果糖。Ir-CHO(邻菲啰啉醛的铱配合物)和Eu-CHO(邻菲啰啉醛的铕配合物)在和谷胱甘肽相关的氨基酸中选择性地识别谷氨酸和含有此片段的谷胱甘肽和金属硫蛋白。Ru-N,N (N,N-二甲基邻菲啰啉的钌配合物)的CH3CN:H2O溶液(9：1,v／v)在12种核糖核酸中以荧光增强的方式选择性地识别ATP。更多还原

【Abstract】 Molecules and ions recognition is an important phenomenon in chemistry, biology and the materials science, and is the basical concept of supramolecular chemistry. The crucial recognition events occur in a much smaller world than that we are accustomed to. Therefore it needs to output the macroscopic signals with the special designed molecular detectors in order to understand and investigate the intermolecular act of recognition. In these signals, the most direct way is expressed by the light signal form, namely visualization. By introducing the special designed fluorescence probes into the target system, it will be able to transform the molecular recognition information into the fluorescence signals to transmit the outside world, thus will cause the communication between the human and the molecular possible and put up a bridge connecting the macrocosm and the microcosm, which is the key point and the hot spot in the current molecule-ion probe research field. The present paper contains following several aspects:1. Cu2+ PET fluorescence probes in living cells:owing to the quenching nature of paramagnetism Cu2+ to the fluorescence molecule excited state, it is a challenging work to design the high selective and sensitive fluorescence enhancement Cu2+ fluorescence probe. The present paper designed and synthesized three Cu2+ fluorescence probes. Probe L1 (bis(2-hydroxyl-naphthalene-carboxaldehyde) benzil dihydrazone) could selectively recognize Cu2+ by the fluorescence quenching way, but Hg2+ had disturbance. Probe L2 was synthesized from the reaction of 6-hydrazino-benzo [de] isoquinoline-1,3-diones with 4-tert-Butyl-2,6-diformyl-phenol. L2 could selectively distinguish Cu2+, and Hg2+ did not disturb it. L2 could be applied in the living cell fluorescence image. As a advancement of such research, by virtue of the electrochemistry survey and the quantum chemistry theoretical calculation, the electronic donor and the acceptor component which match with the Cu2+ redox potential was carefully chosen. The third probe L3, with appropriate front track energy level, was synthesized from the reaction of 7-diethylaminocoumarin-3-aldehyde with benzyl-dihydrazone. The fluorescence of L3 itself was extremely weak. After the Cu2+ coordinating, owing to the orbital energy level reasonable match, the photo induced electron transfer (PET) was blocked. As a result, the fluorescence of L3 obviously increased 75 times. L3 had many merits such as high selectivity, high sensitivity, good water-solubility, and so on. The focuse fluorescence image of living cell of L3 binding Cu2+ indicated L3 was an excellent Cu2+ fluorescence probe in the life system. 2. The design and synthesis of FRET ratio fluorescence probes and entire spectrum fluorescence molecules:It is currently a hot spot in fluorescence sensors research field to design and synthesis dual channel fluorescence probes based on fluorescence resonance energy (FRET) by virtue of fluorescence groups that energy match. In present paper, probes RC1 and RC2 were linked by rhodamine 6G (green fluorescence) or rhodamine B (red fluorescence) group as energy accepter with coumarin (blue fluorescence) group as energy donor through the ethylene diamine group. The selective and competitive experiments indicated the two probes could distinguish Cu2+ through FRET ratio fluorescence. CRB and CR6G were linked by rhodamine B or rhodamine 6G with coumarin through hydrazine group. Such two fluorescence probes could simultaneously distinguish Cu2+ and Hg2+ in aqueous solution by the UV-vis and fluorescence spectra double channels. Based on these probes linked by two fluorophores, the rare earth ion with sharp line emission spectrum was introduced to develop the three fluorescence channel. Single molecule compound RC1-Eu, containing RC1 (containing blue and green fluorophores) and Eu (launching red fluorescence), was synthesized through coordination reaction. By subtly regulating energy transfer process and the efficiency, RC1-Eu could separately launched blue, green and red three-primary colors, and could simultaneously emit blue, green, and red three-primary colors to produce white light when excitated at appropriate wavelength.3. The design and synthesis of fluorescence probes based on metal complexes target biological small molecules:Fluorescence probes target small biological molecules such as monosaccharides, amino acids, ribonucleic acids have important applications in life science and medicine. Based on the pure organic fluorescence probes research, the present paper designed and synthesized a series of fluorescence probes based on fluorescence active Ir, Eu and Ru complexes coordinated with phenanthroline derivatives. Probe Ir-OH (phenanthroline benzene boric acid Ir complex) could selectively recognize fructose in several mono-saccharides by fluorescence quenching approach. Ir-CHO (phenanthroline aldehyde Ir complex) and Eu-CHO (phenanthroline aldehyde Eu complex) could selectively recognize glutamic acid, glutathione and metallothionein by interacting with the particular fragments of them by the N-terminal acylation approach. Ru-N,N (N,N-dimethyl-phenanthroline Ru complex) could selectively distinguish ATP in twelve kinds of ribonucleic acid in CH3CN: H2O (9:1, v/v) solution by fluorescence enhancement response.更多还原