【作者】 郭婷；

【导师】 钱旭红；

【作者基本信息】 华东理工大学 ， 化学生物技术与工程， 2013， 博士

【摘要】 氧化还原过程与信号传导、衰老、疾病等生理过程密切相关,具有重要的研究意义。本论文基于PeT、ICT等荧光探针设计机理,针对活性氧次氯酸、肿瘤细胞的氧化还原过程相关客体,设计合成了几类荧光分子探针。光谱性质及离体水平测试结果表明,这些探针能高灵敏、高选择性识别氧化还原过程相关客体。1.次氯酸根荧光探针：以4-氨基苯酚为受体、1,8-萘酰亚胺为荧光团,设计、合成了两个次氯酸根荧光探针PA和HA。体外荧光测试表明,探针PA是荧光增强型次氯酸探针,而双荧光发射的探针HA可用于活细胞内应用研究。探针HA可通过双峰/单峰发射区分次氯酸根、活性氮以及其它活性氧。活细胞影像结果表明,探针HA具有很好的细胞膜渗透性、低毒性,可用于HeLa细胞中的CIO-检测和成像。2.近红外硝基还原酶及乏氧荧光探针：以尼罗蓝染料为荧光团、对硝基甲酸苄酯为受体,设计、合成了新型近红外乏氧荧光探针NBP。酶水平测试结果表明,探针NBP在NADH存在下可被硝基还原酶催化还原释放荧光团NBF,荧光明显增强。与其它内源性还原物质的竞争性实验进一步证实探针NBP对硝基还原酶的高选择性。NBP透膜性较好,能够对肿瘤细胞A549的乏氧环境进行荧光检测和成像,对实体瘤的早期诊断具有潜在应用价值。3.近红外荧光发色团及pH荧光探针：以半菁染料为荧光团,设计合成了两个新型近红外荧光染料H1和H2。探针HI的pKa为6.1,在pH3.5～7.0范围内紫外响应灵敏。HI在470nm和630nm处实现比色检测,可用于酸性环境pH的检测。探针H2的pKa为5.7,在pH5.0～7.0范围内荧光响应灵敏。酸性条件(pH<5.0)下,探针H2在690nm处有强荧光发射。作为近红外荧光探针,探针H2的pKa与肿瘤细胞或酸性细胞器相匹配,具有活细胞荧光成像的潜在应用价值。4.手性二酸荧光探针：以1,1-联萘为手性识别基团、1,8-萘酰亚胺为荧光团,选用乙二胺或(S,S)-环己二胺为连接臂,设计、合成了两个新型手性荧光探针SP1和SP2。该类探针在500nm处有荧光发射峰。针对手性羧酸,探针SP1和SP2的荧光强度变化不同,其检测手性二酸的潜在应用价值在进一步研究中。更多还原

【Abstract】 Oxidation-reduction (redox) reaction has broad consequences for health, aging and disease and becomes an expanding research area. In this present work, based on the mechanism of photo-induced electron transfer and intramolecular charge transfer, several series of fluorescent probes were developed, which showed selective and sensitive response for guests such as reactive oxygen species, hypoxia and pH.1. Fluorescence probes for hypochloric acid:Two fluorescence probes PA and HA were developed by tethering redox active4-aminophenoxyl to a fluorescent1,8-naphthalimide scaffold. The probe PA showed dramatic fluorescence enhancement toword ClO-.The probe HA exhibited good selectivity against CIO-among various endogenous reactive oxygen or nitrogen species including ONOO-, H2O2, ClO-, O2·-, etc. Importantly, HA displayed a unique two channel ratiometric signal, which renders it advantageous in vitro experiments. HA also exhibited good biocompatibility with its fair aqueous solubility, cell permeability and low cytotoxicity.2. A NIR fluorescence probe for nitroreductase and hypoxia:A Near-infrared fluorescence probe NBP was developed for detection of nitroreductase and hypoxia. Reduction of the p-nitrobenzyl moiety will trigger a cascade reaction to release the Nile Blue fluorophore from a self-collapsible carbamate linkage with a fluorescence enhancement at658nm. A detection limit as low as0.18μg mL-1was calculated. NBP also displayed high reactivity toward other biological reducing agents such as Cys, Hys and GSH. It works well for cell assays and clearly has potentials for in vivo experiments due to its near infrared emission.3. Novel NIR fluorophore and its application in detecion of pH:Two novel pH fluorescence probes H1and H2were designed and synthesized based on hemicyanine dye. Probe H1exhibits a pKa of6.1, while probe H2showed a pKa of5.7. With the agreement of the pKa of H2and pH of both tumor cells or lysosome, H2could be used as a potential NIR fluorescent probe in imaging of acid organelle in cells.4. Fluorescence probes for chiral carboxyli acid:By linking1,8-naphthalimide to chiral (S,S)-1,1-BINOL, we attempted to develop two novel fluorescent probes SP1and SP2for chiral recognition of chiral carboxylic acid. These probes exhibit dual emissions at500nm.更多还原