Towards Treatment of Alzheimer’s Disease by Targeting Pathogenic Tau and Beta-Amyloid Structures

项目摘要 目标1通过基于结构的药物设计解决痴呆症药物的缺乏。这种方法，所以 治疗癌症和艾滋病的成果，是开放的阿尔茨海默氏病（AD），因为进展， 衍射和cryoEM。Tau蛋白的聚集与痴呆的发作密切相关。从 最近从阿尔茨海默病患者的尸检大脑中提取的Tau原纤维的近原子结构， 已经确定了原纤维解聚化合物的药物结合位点（或药效团）。通过 筛选适合药效团的化合物，已经发现了新的Tau解聚剂。这些 解聚剂溶解AD-脑Tau原纤维，但不产生有毒产物。从更多的循环中 Tau原纤维与新的解聚剂的复合物的结构测定，然后进行化合物的结构测定。 筛选、安全和有效的化合物将被寻求来逆转Tau在细胞中的毒性聚集。 个脑袋合成化学家共同研究员加州大学洛杉矶分校教授帕特里克哈伦将合作应用类似的 方法来发现与脑渗透纳米颗粒的解聚剂的复合物。 目的2提出填补Tau和β-聚体的小聚集体结构知识的真空。 淀粉样蛋白，称为低聚物。许多其他研究提供的证据表明，低聚物是更多的 比相同蛋白质的纤维更有细胞毒性。不同纤维形成蛋白的寡聚体具有相同的结构 相似之处在于特定的抗体（A11和M204）识别它们，但不识别它们相应的原纤维。 低聚物的瞬态性质已经击败了以前试图了解其原子结构的尝试，但我们的研究表明， 一个实验室最近发现了一种单克隆Fab，它从AD中提取了相当均一的Tau寡聚体 大脑和稳定他们足够长的时间，使网格适合cryoEM结构确定。 初步的显微照片表明，抗体配体允许β-淀粉样蛋白寡聚体对齐， cryoEM确定结构，随后可用于抑制剂的设计， 解聚剂。 目标3提出在我们的合作研究者加州大学洛杉矶分校的实验室中生长的“迷你大脑”中测试AD药物 教授诺维奇这些类器官是BB的大小，但显示实际的结构和电特性。 人类的大脑它们是由人类细胞制成的，并显示细胞类型和电子信息。 人类的大脑初步工作表明，这些迷你大脑可以感染Tau病理，现在 我们的各种候选药物干扰聚集的Tau的扩散和损伤的能力 可以在其中进行测试。如果成功，这种方法可以为测试阿尔茨海默病药物提供新的途径 在人体试验之前。为了比较，我们的抑制剂和解聚剂也将在小鼠中进行评估。 tau蛋白病模型。

Project Summary Aim 1 addresses the dearth of drugs for dementia, by structure-based drug design. This approach, so fruitful for treating cancer and HIV-AIDS, is opening for Alzheimer’s Disease (AD) because of advances in diffraction and cryoEM. Aggregation of protein Tau is strongly correlated with the onset of dementia. From the recent near-atomic structure of Tau fibrils extracted from the autopsied brain of an Alzheimer’s patient, the drug-binding site (or pharmacophore) has been determined for a fibril-disaggregatng compound. By screening compounds that fit the pharmacophore, new Tau disaggregants have been discovered. These disaggregants dissolve AD-brain Tau fibrils, but do not produce toxic products. From further cycles of structure determination of complexes of Tau fibrils with the new disaggregants, followed by compound screening, safe and effective compounds will be sought to reverse the toxic aggregation of Tau in the brain. Synthetic chemist co-Investigator UCLA Prof. Patrick Harran will collaborate to apply a similar approach to discover complexes of disaggregants with brain-penetrant nanoparticles. Aim 2 proposes to fill the vacuum of knowledge of the structures of small aggregates of Tau and beta- amyloid, known as oligomers. Numerous studies of others provide evidence that oligomers are more cytotoxic than fibrils of the same protein. Oligomers of different fibril-forming proteins share structural similarities in that particular antibodies (A11 & M204) recognize them, but not their corresponding fibrils. The transient nature of oligomers has defeated previous attempts to learn their atomic structures, but our lab has recently discovered a monoclonal Fab that extracts fairly homogeneous oligomers of Tau from AD brains and stabilizes them long enough to make grids suitable for cryoEM structure determination. Preliminary micrographs suggest that antibody ligands permit alignment of beta-amyloid oligomers for cryoEM determination of structure, which may serve subsequently for design of inhibitors and disaggregants. Aim 3 proposes tests of AD drugs in “mini-brains” which are grown in the lab of our co-Investigator UCLA Prof. Novitch. These organoids are the size of a BB yet display structure and electrical properties of actual human brains. They are made from human cells and display the cell types and electrical messaging of human brains. Preliminary work shows these mini-brains can be infected with Tau pathology, and now the ability of our various drug candidates to interfere with the spreading and damage of aggregated Tau can be tested in them. If successful, this approach can provide a new avenue for testing Alzheimer’s drugs prior to human trials. For comparison, our inhibitors and disaggregants will also be assessed in a mouse model of tauopathy.