Novel trityl probes for measurement and bi-modal imaging of superoxide

描述(申请人提供)：超氧化物参与了多种病理生理过程，如缺血再灌注损伤、神经退行性疾病、高血压、衰老和癌症。当超氧化物的生成量很低时，它参与了细胞信号传递过程。然而，在病理过程中产生的大量物质可能会导致细胞功能障碍和组织损伤。因此，超氧化物歧化酶的检测对于我们理解其在正常和病理生理学中的作用至关重要。已经开发了各种分析技术来定量生物体系中的超氧阴离子自由基，包括使用超氧化物还原铁细胞色素c或硝基蓝四氮唑的分光光度方法；基于化学发光和荧光的技术；以及电子顺磁共振(EPR)光谱。然而，这些方法中的许多在测量超氧化物方面都有一定的局限性，主要是缺乏特异性或必要的反应速率。我们已经确定了一类新的基于三苯基的探针，与现有的探针相比，它具有更高的灵敏度和特异度，可以检测和定量生物系统中产生的超氧化物。这些新探针的一个子集是双模式的--它们可以通过EPR和荧光显微镜技术进行检测。因此，这项提议的目标是开发和表征更多用于检测和定量生物系统中超氧化物的新的三苯基探针，并进一步开发和表征用于成像应用的这些探针的双峰子集。因此，提出了以下具体目标：(I)用于生物体系中超氧化物定量的稳定的四氯三苯甲基(PTM)衍生的顺磁自旋探针的合成、表征和验证测试，以及(Ii)用于生物体系中超氧化物分布的定量和荧光成像的稳定的双峰四氯三苯甲基(PTM)衍生的顺磁自旋探针的合成、表征和验证测试。这项建议的目标是开发和表征新的基于三苯基的探针，用于检测、定量和成像生物系统中的超氧化物。超氧化物歧化参与了多种病理生理过程，如缺血再灌注损伤、神经退行性疾病、高血压、衰老和癌症等。在正常情况下，超氧化物的生成量很低，并参与细胞信号传递过程。在病理条件下，大量的超氧化物可能会导致细胞功能障碍和组织损伤。因此，超氧化物歧化酶的检测对于我们理解其在正常和病理生理学中的作用至关重要。

DESCRIPTION (provided by applicant): Superoxide has been implicated in a variety of pathophysiological processes such as ischemia-reperfusion injury, neurodegenerative diseases, hypertension, aging, and cancer. Superoxide, when produced in low amounts, is involved in cell-signaling processes. However, large amounts produced during pathological processes may lead to cellular dysfunction and tissue damage. Hence, the detection of superoxide is critical to our understanding of its role in normal and pathophysiology. A variety of analytical techniques have been developed to quantify superoxide radicals in biological systems including spectrophotometric methods that use the reduction of ferricytochrome c or nitroblue tetrazolium by superoxide; chemiluminescence and fluorescence-based techniques; and electron paramagnetic resonance (EPR) spectroscopy. However, many of these methods have certain limitations for measuring superoxide, primarily lacking specificity or the necessary reaction rates. We have identified a new class of probes based on trityl radicals to detect and quantify superoxide produced in biological systems with greater sensitivity and specificity when compared to existing probes. One subset of these new probes is bi-modal - they can be detected by EPR as well as fluorescence microscopy techniques. Therefore, the goal of this proposal is to develop and characterize additional new trityl-based probes for detection and quantification of superoxide in biological systems, and to further develop and characterize the bi-modal subset of these probes for imaging applications. Accordingly, the following specific aims are proposed: (i) Synthesis, characterization, and validation testing of stable perchloro-triphenylmethyl (PTM)-derived paramagnetic spin probes for quantification of superoxide in biological systems and (ii) Synthesis, characterization, and validation testing of stable, bi-modal perchloro-triphenylmethyl (PTM)-derived paramagnetic spin probes for quantification and fluorescence imaging of superoxide distribution in biological systems. The goal of this proposal is to develop and characterize new trityl-based probes for detection, quantification, and imaging of superoxide in biological systems. Superoxide has been implicated in a variety of pathophysiological processes such as ischemia-reperfusion injury, neurodegenerative diseases, hypertension, aging, and cancer. Under normal circumstances, superoxide is produced in low amounts and is involved in cell-signaling processes. During pathological conditions, large amounts of superoxide may lead to cellular dysfunction and tissue damage. Hence, the detection of superoxide is critical to our understanding of its role in normal and pathophysiology.