Novel Spin Traps for Biological Free Radical Detection

用于生物自由基检测的新型旋转陷阱

• 批准号: 7836983
• 负责人: FREDERICK A. VILLAMENA
• 金额: $ 15.11万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7836983
• 关键词: Anions; Biochemical Reaction; Biological; Biological Models; Cell Line; Cell Membrane Permeability; Cells; Characteristics; Chemicals; Conflict (Psychology); Data; Detection; Development; Disease; Electron Spin Resonance Spectroscopy; Electronics; Free Radical Formation; Free Radicals; Goals; Half-Life; Heart; In Vitro; Kinetics; Lasers; Mediator of activation protein; Methodology; Methods; Mitochondria; Monitor; Organic Synthesis; Oxidative Stress; Oxides; Performance; Physiological; Process; Production; Property; Rattus; Reaction; Reactive Oxygen Species; Reagent; Relative (related person); Reporting; Research Personnel; Role; Site; Specificity; Spin Trapping; Study models; Superoxides; Suspension substance; Suspensions; System; Techniques; Technology; Thermodynamics; Toxic effect; adduct; base; biological systems; biomaterial compatibility; computational chemistry; cytotoxicity; design; flash photolysis; human NOS3 protein; improved; in vivo; innovation; interdisciplinary approach; meetings; nitrone; novel; oxidative damage; programs; pyrroline; stop flow technique

DESCRIPTION (provided by applicant): Over the past two decades, reactive oxygen species have been implicated as critical mediators of oxidative processes and disease mechanisms under physiological conditions. Therefore, it is of critical importance to have a direct technique capable of identifying free radicals at their site of formation in systems ranging from chemical to enzymatic reactions, and cellular to in vivo systems so that the mechanisms and processes underlying the oxidative damage in biological systems can be understood. The overall objective of this proposal is to develop novel spin traps with improved properties that can be applied to study oxidative stress in biological systems using electron paramagnetic resonance (EPR) spectroscopy. The specific monitoring of the formation of biologically relevant reactive oxygen species such as ?OH, O2?-, ROO? or RS? is achieved by the distinctive EPR spectral profile they give after addition to spin traps to form a persistent radical adduct. The most commonly used spin traps, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO) and 5-ethoxycarbonyl-5-methyl-1-pyrroline N-oxide (EMPO), are limited by their poor efficiency of trapping superoxide radical anion, short half-life of the radical adduct formed in biological milieu, cytotoxicity, sensitivity and target specificity. Our aim is to overcome such limitations by employing an interdisciplinary approach in spin trap development that encompasses theoretical prediction, organic synthesis, kinetic determination, toxicity, and ultimately, their experimental application to identify and detect free radical formation in in vitro and in vivo systems at their site of formation. The synergistic application of these technologies will provide an optimal strategy to provide new materials which can be effective in probing the role of reactive oxygen species in biological mechanisms.

描述（由申请人提供）：在过去的二十年中，活性氧已被认为是生理条件下氧化过程和疾病机制的关键介质。因此，拥有一种能够识别从化学到酶反应、从细胞到体内系统等系统中自由基形成位点的直接技术至关重要，以便了解生物系统中氧化损伤的机制和过程可以理解。该提案的总体目标是开发具有改进性能的新型自旋陷阱，可用于使用电子顺磁共振（EPR）光谱研究生物系统中的氧化应激。对生物相关活性氧物质（例如 OH、O2-、ROO？）形成的具体监测还是RS？是通过添加到自旋陷阱中形成持久的自由基加合物后给出的独特的 EPR 光谱轮廓来实现的。最常用的自旋陷阱是5,5-二甲基-1-吡咯啉N-氧化物(DMPO)、二乙氧基磷酰基-5-甲基-1-吡咯啉N-氧化物(DEPMPO)和5-乙氧基羰基-5-甲基-1-吡咯啉N-氧化物（EMPO）受到超氧自由基阴离子捕获效率差、生物环境中形成的自由基加合物半衰期短、细胞毒性、敏感性等限制和目标特异性。我们的目标是通过在自旋陷阱开发中采用跨学科方法来克服这些限制，该方法包括理论预测、有机合成、动力学测定、毒性，以及最终的实验应用，以识别和检测体外和体内系统中自由基的形成。它们的形成地点。这些技术的协同应用将为提供可有效探索活性氧在生物机制中的作用的新材料提供最佳策略。