The Role of Nornicotine in protein Glycation

描述（由申请人提供）：尽管有充分的证据支持烟草滥用的负面影响，但香烟和尼古丁成瘾仍然是美国可预防死亡的主要原因之一。尼古丁对全身和精神的影响都是有据可查的，但很少有研究致力于去尼古丁的生理影响，去尼古丁是一种次要的精神代谢产物。我们认为去甲尼古丁能够参与糖基化蛋白的形成，并且去甲尼古丁在烟草中的天然存在以及作为尼古丁代谢物的存在可能在许多烟草相关疾病状态中具有重要意义。为了探讨去甲尼古丁在蛋白糖基化中的作用，提出了一项多学科研究。首先，利用合成有机化学，我们将确定去甲尼古丁在体外形成Amadori产品的能力。同时，我们将开发一种用于检测nor尼古丁衍生糖化蛋白的快速免疫分析方法。为了促进这一点，我们还将开发新的半抗原，用于制备对这些糖基化产物具有增强特异性的新抗体。该试验将使用长期暴露于尼古丁、去甲尼古丁和去甲尼古丁衍生的Amadori产品的行为大鼠模型进行测试。其次，为了建立Amadori产品引入的主要途径，我们将探索Amadori产品的体内合成速率和该化合物在烤烟中存在的固有比例。我们相信，拟议的研究将提供疾病和烟草滥用之间的分子联系，以及对尼古丁次级代谢物在体内进行化学反应的前所未有的能力的深刻见解。

DESCRIPTION (provided by applicant): Despite ample evidence supporting the negative effects of tobacco abuse, cigarettes and nicotine addiction remain one of the leading causes of preventable death in the United States. Both the systemic and psychoactive effects of nicotine are well-documented, yet little research has been devoted to the physiological effects of nornicotine, a minor psychoactive metabolite. We propose that nornicotine is capable of participating in the formation of glycation proteins, and that the presence of nornicotine both natively in tobacco and as a nicotine metabolite could have significant implications in a number of tobacco-related disease states. In order to explore the role of nornicotine in protein glycation, a multidisciplinary study is proposed. First, using synthetic organic chemistry, we will determine the ability of nornicotine to form Amadori products in vitro. Concurrently, we will develop a rapid immunoassay for the detection of nornicotine-derived glycated proteins. To facilitate this, we will also develop novel haptens for the preparation of new antibodies with enhanced specificity for these glycation products. This assay will be tested using a behavioral rat model of chronic exposure to nicotine, nornicotine, and the nornicotine-derived Amadori product. Second, to establish the major route of introduction of Amadori products, we will explore both the rate of in vivo synthesis of the Amadori product versus the inherent proportion of this compound that exists in cured tobacco. We believe that the proposed studies will provide a molecular link between disease and tobacco abuse as well as a dramatic insight into the unprecedented ability of a secondary nicotine metabolite to perform chemistry in vivo.