Bioorthogonal Generation and Use of Diazo-Compounds

描述（由申请人提供）：生物正交化学反应是研究各种生物途径如何运作的有力工具。这些转化需要反应性的平衡，以达到预期的结果，而不引起有害的副反应。叠氮化物的功能由于其与生物系统的相容性和易于结合而成为这种转化的极好前体。最近，Raines小组开发了一种膦酸酯试剂，能够在温和的条件下在有机溶剂系统中将叠氮化物转化为重氮化合物。第一个目标是合成一种不需要有机溶剂就能发挥作用的相关磷脂酯。该试剂的水溶性和反应性将被系统地优化，以促进所需的转化，而不允许生物系统中存在的水或其他官能团发生反应。当第一个目标实现后，下一步将是探索所得到的重氮化合物的反应性。最后，随着反应性的透彻理解，测试将转移到生物设置。高活性重氮化合物将用于共价交联生物分子，以更好地了解其功能。这只是这种化学转化的许多有用的应用之一。能够在生理条件下从生物相容性叠氮化物中生成高活性重氮化合物的试剂的开发将对化学生物学领域产生重大影响。

DESCRIPTION (provided by applicant): Bioorthogonal chemical reactions are a powerful tool to investigate how various biological pathways operate. These transformations require a balance of reactivity in order to achieve the desired outcome without causing deleterious side reactions. The azide functionality has emerged as an excellent precursor for such transformations due to its compatibility with biological systems and the ease in which it can be incorporated. Recently, the Raines group developed a phosphinoester reagent that was capable of converting azides into diazo compounds under mild conditions in organic based solvent systems. The first goal is to synthesize a related phosphinoester that is able to function without the need of organic solvents. The water solubility and reactivity of the reagent will be systematically optimized to promote the desired transformation without allowing water or other functional groups present in biological systems to react. When the first goal has been realized, the next step will be to explore the reactivity of the resulting diazo compounds. Finally, with a thorough understanding of reactivity, the tests will be moved to a biological setting. The highly reactive diazo compounds will be used to covalently crosslink biomolecules to better understand their function. This is just one of many useful applications of this chemical transformation. The development of a reagent capable of producing highly reactive diazo compounds from biologically compatible azides under physiological conditions will have a significant impact on the field of chemical biology.