INTERDISCIPLINARY APPROACH TO ALZHEIMER DRUG DISCOVERY

阿尔茨海默病(AD)是一种以沉积为特征的 脑血管和脑实质β/A4-淀粉样蛋白和进展性 有效的胆碱能神经传递不足。这两个功能 是拟议的药物发现小组的重点。 项目1的研究人员将专注于淀粉样前体蛋白(APP) 在大脑和脑血管中处理，使用合成的APP多肽和 重组APP全蛋白和APPase片段将被提纯并 特色化的。除了这些酶学研究外，APP处理 将在培养的内皮细胞、平滑肌细胞和 血小板。 项目2中的调查人员还将使用完整的APP处理测试 培养细胞，根据细胞生物学合理选择化合物 并筛选这些化合物在调节APP处理方面的有效性。 在存在先导化合物的地方，将对衍生品进行检查，以便 确定不同类别的结构-活性关系 化合物。多种宿主细胞系统和在体脑组织中的检测 自由移动的啮齿动物将被用作屏蔽物。补充细胞 项目1和2的生物学和生化方法，研究人员在 项目3将寻求大脑淀粉样变的活体模型， 在转基因小鼠中过表达突变APP可能有利于 淀粉样变。 进行性胆碱能神经传递缺陷是另一个原因 AD的突出特点。精细分子和生理特性 胆碱能系统构成了项目3的另一个重点，是 项目4的主要关注点是新开发的区分 脑M受体亚型将用于分析水平和 亚型在基底前脑、海马区和 老年人的新皮质。同样沿着这条线，项目4将提供 关于可能的治疗机会的详细信息 神经肽和功能性拮抗剂甘丙肽的操作 胆碱能神经传递。甘氨酸能在阿尔茨海默病中的保存 系统，再加上胆碱能神经元的丧失，表明甘丙素 拮抗剂可能在临床上有用。 一个核心基金将为以下项目提供科学支持 药物发现小组。在核心的职能中， 提供重组野生型和突变型APP和APP片段， 合成肽、蛋白质纯化和测序以及工业 负责化合物衍生和大规模合成的联络工作。联络： 此外，还将提供法律和专利事务以及便利 小组所有成员之间及时沟通的重要性。

Alzheimer disease (AD) is a characterized by the deposition of cerebrovascular and cerebral parenchymal beta/A4-amyloid and progressive deficiency of effective cholinergic neurotransmission. These two features are the focus of the proposed Drug Discovery Group. Investigators in Project 1 will focus on amyloid precursor protein (APP) processing in brain and cerebral vessels, using synthetic APP peptides and recombinant APP holoprotein and fragments as APPases will be purified and characterized. In addition to these enzymological studies, APP processing will be studied in cultured endothelial cells, smooth muscle cells and platelets. Investigators in Project 2 will also use an APP processing assay in intact cultured cells, rationally choosing compounds based on the cell biology of APP and screening the compounds for efficacy in regulating APP processing. Where lead compounds exist, derivatives will be examined in order to determine structure-activity relationships of the various classes of compounds. Various host cell systems and an in vivo assay in the brain of the freely moving rodent will be used as screens. Complementing the cell biology and biochemical approaches of Projects 1 and 2, investigators in Project 3 will seek an in vivo model of cerebral amyloidogenesis, overexpressing in transgenic mice mutant APPs which are likely to favor amyloidogenesis. Progressive deficiency in cholinergic neurotransmission is another prominent feature of AD. Fine molecular and physiological characterization of the cholinergic system forms another focus of Project 3 and is the primary focus of Project 4. Newly developed antibodies that differentiate brain muscarinic receptor subtypes will be used to analyze levels and distributions of subtypes in the basal forebrain, hippocampus, and neocortex of aged humans. Also along this line, Project 4 will provide details regarding the possible therapeutic opportunity offered by manipulation of galanin, a neuropeptide and functional antagonist of cholinergic neurotransmission. The preservation in AD of galaninergic systems, coupled with loss of cholinergic neurons, suggests that galanin antagonists might be clinically useful. A Core Facility will serve to provide scientific support to the Projects of the Drug Discovery Group. Among the functions of the Core will be the provision of recombinant wildtype and mutant APP and APP fragments, synthetic peptides, protein purification and sequencing, and industrial liaison for compound derivatization and large-scale synthesis. Liaison for legal and patent matters will also be provided as well as the facilitation of timely communication among all members of the Group.