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

描述（由申请人提供）：超氧化物涉及多种病理生理过程，如缺血再灌注损伤、神经退行性疾病、高血压、衰老和癌症。少量产生的超氧化物参与细胞信号传递过程。然而，在病理过程中大量产生可能导致细胞功能障碍和组织损伤。因此，超氧化物的检测对我们理解其在正常和病理生理中的作用至关重要。各种各样的分析技术已经发展到量化生物系统中的超氧化物自由基，包括分光光度法，使用超氧化物还原铁细胞色素c或硝基蓝四唑；化学发光和荧光技术；电子顺磁共振（EPR）光谱学。然而，许多这些方法在测量超氧化物有一定的局限性，主要是缺乏特异性或必要的反应速率。与现有探针相比，我们已经确定了一类基于三烷基自由基的新型探针，用于检测和量化生物系统中产生的超氧化物，具有更高的灵敏度和特异性。这些新探针的一个子集是双模态的——它们可以通过EPR和荧光显微镜技术检测到。因此，本提案的目标是开发和表征其他新的基于三烷基的探针，用于检测和定量生物系统中的超氧化物，并进一步开发和表征这些探针的双模态子集，用于成像应用。因此，提出了以下具体目标：(i)合成、表征和验证测试稳定的高氯三苯基甲基（PTM）衍生顺磁自旋探针，用于定量生物系统中的超氧化物；（ii）合成、表征和验证测试稳定的双峰高氯三苯基甲基（PTM）衍生顺磁自旋探针，用于定量和荧光成像生物系统中的超氧化物分布。本提案的目标是开发和表征新的基于三烷基的探针，用于生物系统中超氧化物的检测、定量和成像。超氧化物参与多种病理生理过程，如缺血再灌注损伤、神经退行性疾病、高血压、衰老和癌症。在正常情况下，超氧化物的产生量很低，并参与细胞信号传导过程。在病理状态下，大量的超氧化物可导致细胞功能障碍和组织损伤。因此，超氧化物的检测对我们理解其在正常和病理生理中的作用至关重要。

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.