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活性类似物异丙酚，促进我们对全身麻醉剂异丙酚药理学的理解。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，异丙酚和其他全身麻醉剂越来越被认为是神经发育和神经退行性风险的介质，具有引起不可逆表型的潜力。间偶氮异丙酚含有一个重氮嘧啶部分，当暴露在长波紫外线（~370 nm）下时，该部分会发生光解，产生反应性的二氧化碳中间体。在体内局部刺激异丙酚的新技术将阐明导致这种全身麻醉产生的无意识的大脑区域。meta-azi-异丙酚的综合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元偶氮异丙酚的蛋白质结合位点将提供结合特异性的见解，从而提高我们对这些小疏水分子药物作用的理解。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中的广泛影响的认识，并将为未来的研究创造领域，以尽量减少这些广泛使用的药物的有害潜力。

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.