Light-enabled identification of the neural substrates for alkylphenol anesthesia

描述（由申请人提供）：本项目旨在通过使用光敏类似物meta-azi-propofol，帮助我们了解全身麻醉剂丙泊酚的药理学。异丙酚诱导催眠的机制在分子和系统神经科学水平上仍不清楚。此外，丙泊酚和其他全身麻醉药越来越被认为是神经发育和神经退行性风险的介质，可能导致不可逆的表型。间-阿齐-丙泊酚含有一个二氮杂环丙烯部分，当暴露于长波紫外光（~370 nm）时发生光解，产生一种反应性卡宾中间体。新的技术，涉及局部刺激的间阿齐-丙泊酚在体内将阐明大脑区域，有助于无意识产生的全身麻醉。间-azi-丙泊酚的全面体外蛋白质组学实验将揭示可能导致烷基酚麻醉的分子底物以及与这些麻醉化合物相关的急性和长期副作用。此外，通过光标记和串联质谱技术鉴定间-azi-丙泊酚的蛋白质结合位点将提供对结合特异性的了解，以提高我们对这些小疏水分子的药物作用的理解。通过解决这些目标，所获得的知识将增加我们对全身麻醉药在中枢神经系统内的广泛影响的认识，并将为未来的研究创造领域，以最大限度地减少这些广泛使用的药物的有害潜力。 公共卫生相关性：每年，美国和国外都有数百万人使用全身麻醉剂，尽管人们对控制其临床用途的机制有着清楚的了解。此外，关于这些化合物引起年轻人中枢神经系统发育缺陷和加速老年人神经退行性过程的能力的担忧也在增加。目前可用药物的最佳使用和改进的全身麻醉药的开发的前景依赖于我们辨别这些化合物的药理学的能力。

DESCRIPTION (provided by applicant): This project serves to contribute to our understanding of the pharmacology of the general anesthetic propofol through the use of a photoactive analogue, meta-azi-propofol. The mechanisms of propofol-induced hypnosis remain unclear on both molecular and systems neuroscience levels. Further, propofol and other general anesthetics are becoming increasingly recognized as mediators of neurodevelopmental and neurodegenerative risks with the potential for causing irreversible phenotypes. Meta-azi-propofol contains a diazirine moiety that undergoes photolysis when exposed to long wave ultraviolet light (~370 nm) to create a reactive carbene intermediate. Novel techniques involving localized stimulation of meta-azi-propofol in vivo will elucidate brain regions that contribute to the unconsciousness produced by this general anesthetic. Comprehensive in vitro proteomic experiments with meta-azi-propofol will uncover molecular substrates that potentially contribute to alkylphenol anesthesia and the acute and long-term side effects associated with these anesthetic compounds. Further, the identification of protein binding sites of meta-azi-propofol through photolabeling and tandem mass spectrometry techniques will provide insight into binding specificity in order to improve our understanding of drug action by these small hydrophobic molecules. By addressing these aims, the knowledge gained will add to our appreciation for the widespread impact of general anesthetics within the central nervous system and will generate areas for future research to minimize the harmful potential of these widely used drugs. PUBLIC HEALTH RELEVANCE: Every year general anesthetics are administered to millions of people in America and abroad despite a clear understanding of the mechanisms that govern their clinical usefulness. Additionally, concern has risen regarding the ability of these compounds to cause developmental deficiencies in the central nervous system of the young and to accelerate neurodegenerative processes in the elderly. The optimal usage of currently available drugs and the prospect for the development of improved general anesthetics relies on our ability to discern the pharmacology of these compounds.