Organometallic Sensors for Cellular Small Molecule Detection

7.项目摘要/摘要 非生物过渡金属催化剂在生物体系中的应用经历了 近年来，从人造金属酶到分子的例子都有显著的增长 用于检测一氧化碳等具有挑战性的分析物的探头。私家侦探有长期利益关系 发现、发展和理解检测具有挑战性的生物学相关性的策略 分子通过过渡金属独特的反应性。最近，Michel组织报告说，BODIPY 乙烯探针(BEP)作为第一个基于前荧光活性的传感器(ABS)用于检测 生物系统中的乙烯。这些ABS采用了著名的Ru-烯烃歧化催化剂， 都很容易与乙烯反应。 虽然乙烯长期以来一直被认为是一种重要的植物激素，但它也被证明 在哺乳动物中由于氧化应激而产生，而氧化应激是多种疾病的标志。特别是 乙烯产生于过氧化脂质和/或形成过氧化脂质的中间体的自由基碎裂。 过氧化脂质的形成是活性氧物种的结果，这与比赛有关。 压力或信号在许多疾病中的作用，包括癌症、心血管疾病和 神经退行性疾病等。虽然有一些复杂的光谱方法 为了灵敏地测量呼气中的生物标记物乙烯，这些方法是必要的 在空间分辨率和样本复杂性方面受到限制。自从报告了我们最初的ABS方法以来，我们的团队 进行了系统的配基调节研究，以提高探针的响应时间和灵敏度。 通过这项工作，探测极限提高了近两个数量级。尽管如此 在内源乙烯检测中的广泛应用仍然是一个重要的问题 与在生物系统中保持稳健稳定性的同时进一步提高灵敏度的修改有关。 开发这项技术的下一阶段将建立在机械洞察和最新进展的基础上 在催化烯烃歧化反应中。这将通过跨学科的综合方法来实现， 分析和光物理表征，以及亚细胞定位研究。据预计， 拟议的研究将导致高度敏感的乙烯ABS定位于亚细胞位置 预计将在浓度最高的地方发现乙烯。此外，我们还希望探索小说 乙烯检测策略超越了通常用于乙烯和 其他小分子分析物。正如过去10-15年的情况一样，我们预计将有 在蜂窝环境中运行的过渡金属催化剂的持续增长 原本不可能实现的功能。拟议方案的研究和概念将 继续为这一领域作出重大贡献。 1

7. Project Summary/Abstract The adaptation of abiotic transition metal catalysts for applications in biological systems has undergone remarkable growth in recent years with examples ranging from artificial metalloenzymes to molecular probes for detection of challenging analytes such as carbon monoxide. The PI has a standing interest in discovery, development, and understanding of strategies for detecting challenging biologically relevant molecules via the unique reactivity of transition metals. Most recently, the Michel group reported BODIPY Ethylene Probes (BEPs) as the first profluorescent Activity Based Sensors (ABS) for the detection of ethylene in biological systems. These ABS adapted well-known ruthenium olefin metathesis catalyst, which are known to readily react with ethylene. While ethylene has long been known as an important plant hormone it has also been demonstrated to be produced in mammals as a result of oxidative stress that is hallmark to numerous diseases. In particular ethylene arises from the radical fragmentation of lipid peroxides and/or intermediates in their formation. The formation of lipid peroxides is a result of reactive oxygen species, which are implicated as playing stress or signaling roles in numerous diseases including cancer, cardiovascular disease, and neurodegenerative diseases amongst others. While there are some sophisticated spectroscopic methods for sensitively measuring the biomarker ethylene in exhaled breath, these approaches are necessarily limited in spatial resolution and complexity of sample. Since reporting our initial ABS approach, our group has conducted systematic ligand modulation studies to improve probe response time and sensitivity. Through this work the limit-of-detection was improved nearly two orders of magnitude. Despite these advancements, important questions remain for broad applications in the detection of endogenous ethylene related to modifications that further improve sensitivity while retaining robust stability in biological systems. The next stage of developing this technology will build on mechanistic insight and recent advancements in catalytic olefin metathesis. This will be accomplished through an interdisciplinary approach of synthesis, analytical and photophysical characterization, and subcellular localization studies. It is anticipated that the proposed research will result in highly sensitive ethylene ABS localized to subcellular locations where ethylene is expected to be found in the highest concentrations. Further we expect to explore novel strategies for ethylene detection beyond the dosimetric responses generally employed for ethylene and other small molecule analytes. As has been the case over the past 10-15 years, we envision that there will be continued growth of transition metal catalysts operating in cellular environments to perform critical functions that would otherwise not be possible. The research and concepts of the proposed program will continue to significantly contribute to this field. 1