Light-enabled identification of the neural substrates for alkylphenol anesthesia

描述（由申请人提供）：该项目通过使用光活性类似物间阿齐-丙泊酚，有助于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，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，有可能导致不可逆的表型。间阿齐-丙泊酚含有二氮丙啶部分，当暴露于长波紫外线（~370 nm）时会发生光解，产生反应性卡宾中间体。涉及体内局部刺激间阿齐-丙泊酚的新技术将阐明导致这种全身麻醉产生无意识的大脑区域。使用间阿齐-丙泊酚进行的综合体外蛋白质组学实验将揭示可能有助于烷基酚麻醉以及与这些麻醉化合物相关的急性和长期副作用的分子底物。此外，通过光标记和串联质谱技术鉴定间阿齐-丙泊酚的蛋白质结合位点将提供对结合特异性的深入了解，以提高我们对这些疏水性小分子药物作用的理解。通过实现这些目标，所获得的知识将加深我们对全身麻醉药在中枢神经系统内的广泛影响的认识，并将为未来的研究领域提供新的领域，以最大限度地减少这些广泛使用的药物的潜在危害。

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.