Generation and Solution of Reactivity of Nitroxyl (HNO)

化学结构、动力学和机制项目支持约翰霍普金斯大学化学系的John P. Toscano教授对硝基（HNO）的生成和溶液反应性的研究，这一研究最近受到了极大的关注，特别是作为目前治疗心力衰竭的潜在替代方法。由于其固有的反应性，HNO必须在原位生成，但目前只有非常有限数量的合适前体存在。目前还没有一种可行的方法可以在溶液或生物相关制剂中直接检测HNO，这增加了研究HNO化学的难度。提出的研究将解决当前在理解HNO化学和生物化学、可用的HNO供体分子和实际有用的直接HNO检测技术方面的不足。特别是，研究将开发(1)新的HNO前体以及(2)以膜入口质谱（MIMS）为重点的HNO检测和研究的新分析方法。此外，研究还将解决HNO诱导的磷酸化蛋白（PLN）的修饰，该蛋白参与调节心脏肌浆网钙泵（SERCA2a），以确定HNO增强SERCA2a活性的机制。新的HNO供体的开发，结合反应性研究和HNO诱导蛋白修饰的评估，不仅将为HNO的生物活性提供基本的见解，而且可能最终导致开发一类用于治疗心力衰竭的新型化合物。此外，关于开发新的HNO前体、检测HNO的新分析工具以及阐明HNO诱导的PLN修饰的拟议研究将为研究生和本科生提供广泛的培训。它将涉及要研究的前体的合成、反应混合物的仔细分析、产品的定量、时间分辨光谱研究、生化分析和现代分析技术的使用。学生将获得广泛的技能，使他们能够解决复杂的问题，并将有利于未来在工业和学术界的应用。在本资助期内，将继续积极鼓励本科生参与研究，以及努力扩大妇女和人数不足群体的参与。本科生，无论是来自约翰霍普金斯大学或附近的大学/学院的小或没有博士课程，将继续参与研究在学年和/或在夏季的几个月。

The Chemical Structure, Dynamics, and Mechanism Program supports Professor John P. Toscano of the Chemistry Department at Johns Hopkins University for research involving the generation and solution reactivity of nitroxyl (HNO), which has recently received significant attention, especially as a potential alternative to current treatments of cardiac failure. Due to its inherent reactivity, HNO must be generated in situ, but only a very limited number of appropriate precursors currently exist. Adding to the difficulty of studying HNO chemistry is that a viable method for its direct detection in solution or biologically relevant preparations is not currently available. The proposed research will address these current deficiencies in the understanding of HNO chemistry and biochemistry, in available HNO donor molecules, and in practically useful direct HNO detection techniques. In particular, research will develop (1) new precursors to HNO as well as (2) new analytical approaches for the detection and study of HNO focusing on membrane inlet mass spectrometry (MIMS). In addition, research will also address the HNO-induced modifications of the protein phospholamban (PLN), which is involved in regulating the cardiac sarcoplasmic reticulum calcium pump (SERCA2a), to determine the mechanism by which HNO enhances SERCA2a activity.The development of new HNO donors, combined with reactivity studies and the evaluation of HNO-induced protein modifications, will not only provide fundamental insight into the biological activity of HNO, but may also ultimately lead to the development of a new class of compounds for the treatment of heart failure. In addition, proposed research on the development of new precursors to HNO, new analytical tools for its detection, and elucidation of HNO-induced modifications of PLN will afford broad training for graduate students and undergraduates. It will involve the synthesis of the precursors to be studied, careful analysis of reaction mixtures, quantification of products, time-resolved spectroscopic studies, biochemical analyses, and the use of modern analytical techniques. Students will acquire a wide range of skills that will allow them to solve complex problems and will be beneficial for future applications both in industry and academia. Undergraduate participation in research, as well as efforts to broaden the participation of women and underrepresented groups, will continue to be actively encouraged in the current funding period. Undergraduates, either from Johns Hopkins University or nearby universities/colleges with small or no Ph.D. programs, will continue to be involved in research during the academic year and/or over the summer months.