The Role of Nornicotine in protein Glycation

描述(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的死亡的主要原因之一。尼古丁对全身和精神的影响都有很好的文献记载，但很少有研究致力于去甲尼古丁的生理影响，去甲尼古丁是一种次要的精神活性代谢物。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存在以及作为尼古丁代谢物的存在可能在一些与烟草相关的疾病状态中具有重要意义。为了探索去甲尼古丁在蛋白质糖基化中的作用，提出了一项多学科的研究。首先，利用合成有机化学，我们将测定去甲尼古丁在体外形成Amadori产物的能力。同时，我们将开发一种快速免疫分析方法来检测去甲尼古丁衍生的糖化蛋白。为了促进这一点，我们还将开发新的半抗原来制备具有增强这些糖基化产物特异性的新抗体。这项测试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，为了确定引入Amadori产品的主要途径，我们将探索Amadori产品在体内合成的速度与这种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作用的前所未有的能力的戏剧性洞察。

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.