Cholinesterase Inhibitors in Alzheimer's Disease

项目摘要 阿尔茨海默病(AD)的特征是胆碱能功能障碍和淀粉样多肽的沉积 (a？)源自A？前体蛋白(APP)。目前FDA批准的四种治疗AD的药物属于这一类别 胆碱酯酶抑制剂(ChEI)。这些药物对治疗轻至中度AD很有用，但使用有限 在高级阶段。我们的目标是测试新的乙酰胆碱酯酶抑制剂(AChEIs)或丁酰胆碱酯酶 抑制物(BChEIs)在AD的病理生理学中的关键步骤，以更好地了解药物的效果。 我们的重点是测试新的治疗策略，以验证和新的疾病靶点。我们的假设是 某些CHEI具有神经保护活性，不依赖于它们的胆碱能活性，这是因为它们降低了A？ ？财产。我们提出的研究ChEI药物在淀粉样变通路上的机制的建议是基于我们的 结果某些CHEIs可显著降低A？在培养的细胞和动物中的水平。具体目标是： [目的]研究新型AChEIs和BChEIs对APP通路阶跃的影响。我们将研究i)的影响 APP途径上的新ChEIs：A？和BACE水平，II)官能团取代的影响，III) APPmRNA5‘-非编码区，以及iv)突触蛋白。2]检测一组新的CHEI对A？沉积 在活体内。我们将研究不同剂量的选择性AChEIs和BChEIs对APP和A？成双的 转基因APPswe-Tau淀粉样斑块加缠结小鼠。3]研究胆碱酯酶在 疼痛基因敲除小鼠。我们将研究AchE酶在APP上的作用。多肽与突触蛋白 来自新型AChE基因敲除小鼠的脑组织样本中的标志物。4]调查CHEIs对以下方面的影响 人类(存档)样本中的分子标记。为了验证动物研究，我们将测试 APP和A对精选CHEIs的治疗？在人体血浆和/或脑脊液样本中的水平。 主要终点是定量的和功能性的：细胞存活率、酶分析和APP途径水平 蛋白质/多肽和突触蛋白标记物。我们将机械地选择与 ChE的外周变构结合结构域与酯类和阴离子结合结构域并在 细胞培养和动物模型。这项工作将表明CHEIs对A？和突触蛋白， 与它们对酶的选择性无关。这一特性将有助于最大限度地发挥其在 淀粉样蛋白和突触蛋白，可用于设计更好的阿尔茨海默病治疗剂。项目叙事 阿尔茨海默病(AD)的特征是胆碱能系统的突触前标志物减少， 尤其是大脑中与记忆和学习有关的区域，以及淀粉样多肽的沉积。 (a？)，它是由A？前体蛋白(APP)。FDA目前批准的治疗AD的五种药物中的四种 是胆碱酯酶抑制剂(ChEI)，它通过抑制乙酰胆碱(ACh)增加大脑的供应 胆碱酯酶(ChE)的酶，从而保护胆碱能回路，这被认为是中介记忆 小路。相当令人惊讶的是，这些药物对记忆没有实质性的影响，但有 稍微延长一段时间保存认知的意想不到和可喜的结果。现在 该提案试图通过CHEIs和FINE来表征神经保存和保护的机制 调整这一途径以改善CHEIs的这一有益属性。我们的目标是测试新的乙酰胆碱酯酶 抗胆碱酯酶抑制剂(AChEIs)或丁酰胆碱酯酶抑制剂(BChEIs)在糖尿病的病理生理学中的关键步骤 为了更好地了解药物的效果。我们建议对CHEI药物的作用机制进行研究。 将APP加工成潜在毒性A？的淀粉样变通路。我们的研究最初集中在 这些药在A上吗？最近已经扩展到突触蛋白标记物和其他 神经保护作用。该提案的结果是确定CHEIs阻止的机制 潜在的有毒物质A？并利用这一特性开发新的治疗剂。

PROJECT ABSTRACT Alzheimer's disease (AD) is characterized by cholinergic dysfunction and depositions of the amyloid ?-peptide (A?) derived from A? precursor protein (APP). Four current FDA-approved drugs for AD belong to the category of cholinesterase inhibitor (ChEI). These drugs are useful for treatment of mild to moderate AD but limited use in advanced stages. Our goal is to test novel acetylcholinesterase inhibitors (AChEIs) or butyrylcholinesterase inhibitors (BChEIs) against pivotal steps in the pathophysiology of AD to better understand the drugs' effects. Our focus is to test new therapeutic strategies to both validated and novel disease targets. Our hypothesis is that certain ChEIs have neuroprotective activity independent of their cholinergic activity due to their A?-lowering ? property. Our proposal to study the mechanism of ChEI drugs on amyloidogenic pathways is based on our results that certain ChEIs significantly reduced A? levels in cultured cells and animals. The specific aims are: 1] To study the effect of novel AChEIs and BChEIs on APP pathway steps. We will examine i) the effect of novel ChEIs on APP pathway: A? and BACE levels, ii) the effect of substitution of functional groups, iii) APPmRNA 5'-UTR, and iv) synaptic proteins. 2] To test the effect of a novel group of ChEIs on A? deposition in vivo. We will study the effects of different doses of selective AChEIs and BChEIs on APP and A? in double transgenic APPSWE-Tau amyloid plaque plus tangle producing mice. 3] To examine the role of ChE enzyme in AChE knockout mice. We will study the role of the AChE enzyme on APP, A? peptides and synaptic protein markers in the brain tissue samples from novel AChE knockout mice. 4] To investigate the effect of ChEIs on molecular markers in human (archived) samples. To validate the animal studies, we will test the effect of treatment of selected ChEIs on APP and A? levels in human plasma and/or CSF samples. The primary end-points are quantitative and functional: Cell survival, enzyme assay and levels of APP pathway protein/peptides and synaptic protein markers. We will mechanistically select ChEIs that interact with the peripheral allosteric binding domain of ChE and with the esteratic and anionic binding domains and test them in cell culture and animal models. This work will indicate unique effect of ChEIs on A? and synaptic proteins, independent of their selectivity for the enzyme. This property will help maximize their beneficial effects on amyloid and synaptic proteins, which can be utilized to design better therapeutic agents for AD. PROJECT NARRATIVE Alzheimer's disease (AD) is characterized by a reduction in the presynaptic markers of the cholinergic system, particularly in areas of the brain related to memory and learning, and by depositions of the amyloid ?¿peptide (A?), which is derived from the A? precursor protein (APP). Four of five current FDA-approved drugs for AD are cholinesterase inhibitors (ChEI), which increase the brain's supply of acetylcholine (ACh) by inhibiting cholinesterases (ChE) enzyme and thus preserve cholinergic circuits, which are believed to mediate memory pathways. Rather surprisingly, these drugs did not have a substantial effect on memory, but had an unexpected and welcome outcome of preserving cognition for a slightly extended period of time. The present proposal attempts to characterize the mechanisms of neuropreservation and protection by the ChEIs and fine tune this pathway to improve this beneficial property of ChEIs. Our goal is to test novel acetylcholinesterase inhibitors (AChEIs) or butyrylcholinesterase inhibitors (BChEIs) against pivotal steps in the pathophysiology of AD to better understand the effects of the drugs. We propose to study the mechanism of ChEI drugs on amyloidogenic pathways that process APP to potentially toxic A?. Our studies initially focused on the effects of these drugs on A? production and have recently been extended to synaptic protein markers and other neuroprotective effects. The outcome of the proposal is to identify mechanisms by which ChEIs block potentially toxic A? levels and to utilize this property in developing novel therapeutic agents.