A Simple and Robust Indicator for Glutathione

慢性病是世界上主要的死亡原因，占所有死亡人数的60%以上。 众所周知，三肽谷胱甘肽（GSH）水平降低与慢性 疾病，并且在监测疾病进展和治疗干预中感兴趣。为 例如，最近的研究表明，监测全血谷胱甘肽（GSH）水平可能有益 用于诊断无症状心力衰竭和线粒体疾病GSH监测也是 可用于慢性肾病或心脏手术后的健康评估 手术然而，健康受试者和患者中报告的GSH水平均表现出实验室间差异。 变化量血浆GSH水平经常报告，尽管事实上，他们占不到0.5% 循环的GSH。不同的样品处理和结合步骤在实验室间传播 这是因为它们导致不同程度的样品氧化和标记。为了实现 一致和有效的GSH监测，这项建议的主要目标是开发一种荧光 直接在最低稀释的全血中起作用的指示剂。为了实现这一目标， 建议的具体目标如下： 1. NIR活性GSH选择性指示剂的设计、合成和评价。 基于上述初步数据，我们的工作假设是， GSH的指示剂可以设计成通过多点超分子相互作用结合GSH， 产生伴随的荧光信号，而没有来自血红蛋白或其它 血液成分。 2.开发全血中GSH的检测和定量方法。 我们假设，再次根据我们的初步数据，谷胱甘肽的选择性和敏感性， 在疾病相关的范围内，可以通过GSH选择性的设计在人类血液中实现 含有NIR活性探针的受体。因此，为实现这一目标而提出的研究重点是 优化和示范的效用所提出的设计原则和综合 通过广泛的评估和验证研究确定指标。

Chronic diseases are the leading cause of death in the world, accounting for over 60 % of all fatalities. Decreased levels of the tripeptide glutathione (GSH) are well-known to be associated with chronic diseases and are of interest in monitoring disease progression and therapeutic interventions. For example, recent studies show that monitoring whole blood glutathione (GSH) levels can be of benefit towards diagnosing asymptomatic heart failure and mitochondrial disorders. GSH monitoring is also useful in the health assessment of patients with chronic renal disease or post-operative open heart surgery. However, reported levels of GSH in both healthy subjects and patients exhibit inter-laboratory variation. Plasma GSH levels are often reported despite the fact that they account for less than 0.5 % of the circulating GSH. Divergent sample processing and conjugation steps propagate inter-laboratory variability, as they result in varying degrees of sample oxidation and labeling. In order to achieve consistent and efficient GSH monitoring, the major goal of this proposal is to develop a fluorescent indicator that functions directly in minimally-diluted whole blood. In order to achieve this goal, the following specific aims are proposed: 1. Design, synthesis and evaluation of NIR-active GSH-selective indicators. Based on the preliminary data referred to above, our working hypothesis is that highly selective indicators for GSH can be designed to bind GSH via multipoint supramolecular interactions and produce concomitant fluorescence signals without optical interference from hemoglobin or other blood components. 2. Develop methodology for detection and quantitation of GSH in whole blood. We postulate, again on the basis of our preliminary data, that selectivity and sensitivity for GSH over a disease-relevant range can be achieved in human blood via the design of GSH-selective NIR active probe-containing receptors. The studies proposed in this aim are thus focused on optimization and demonstration of the utility of the proposed design principles and synthetic indicators via extensive evaluation and validation studies.