Discovery of apoE4 modulators for Alzheimer’s disease therapy

发现用于治疗阿尔茨海默病的 apoE4 调节剂

• 批准号: 10502511
• 负责人: THOMAS D Y CHUNG
• 金额: $ 46.79万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10502511
• 关键词: 3-Dimensional; Abeta synthesis; Actins; Affect; Age-Years; Alleles; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease risk; Alzheimer' s disease therapeutic; Alzheimer' s disease therapy; Animals; Apolipoprotein E; Astrocytes; Benchmarking; Biochemical; Biological; Biological Assay; Blood - brain barrier anatomy; Brain; CD69 antigen; Catalogs; Cell model; Cells; Cerebrum; Chemicals; Chemistry; Cognitive deficits; Collaborations; Complement; Critical Pathways; Dementia; Detection; Disease; Down-Regulation; Drug Kinetics; Evaluation; Future; Genes; Genotype; Human; Late Onset Alzheimer Disease; Lead; Life; Luciferases; Measures; Modeling; Monitor; Morphologic artifacts; Neurofibrillary Tangles; Neurons; Organoids; Pathogenicity; Pathologic; Penetrance; Penetration; Performance; Pharmaceutical Preparations; Phenotype; Positioning Attribute; Property; Protein Fragment; Proteins; Reporter; Risk; Rodent Model; Senile Plaques; Signal Transduction; Synaptic plasticity; Tauopathies; Therapeutic Effect; Toxic effect; Treatment Efficacy; abeta accumulation; amyloid pathology; analog; apolipoprotein E-4; base; cheminformatics; cytotoxic; cytotoxicity; dementia risk; drug candidate; drug discovery; genetic risk factor; high throughput screening; improved; in vivo; induced pluripotent stem cell; inhibitor; neuronal survival; novel; novel therapeutic intervention; novel therapeutics; pre-clinical; programs; response; scaffold; scale up; small molecule; small molecule libraries; tau Proteins; tau phosphorylation; therapeutic target

PROJECT SUMMARY Alzheimer's disease (AD) is the most common form of dementia accompanied by detrimental cognitive deficits and pathological accumulation of amyloid-β (Aβ) plaques and tau-containing neurofibrillary tangles. The majority of AD cases occur late in life, and it usually develops after 65 years of age. The strongest genetic risk factor for late-onset AD (LOAD) is apolipoprotein E (APOE) genotype, with the ε4 allele increasing AD risk and the ε2 allele being protective compared with the common ε3 allele. Increasing evidence indicates that down- regulation of apoE4 protein level and/or inhibition of apoE4 aggregation not only alleviate amyloid pathology but also protect against tau-mediated neurodegeneration. In our preliminary studies, we have developed an apoE4 reporter assay with a split luciferase protein-fragment complementation, enabling us to not only monitor apoE4 protein level but also measure apoE4 self-oligomerization/aggregation by luciferase bioluminescent signals in a high-throughput screening format. Under the support of the Mayo-SBP (Sanford Burnham Prebys) Drug Discovery Collaboration Program, we performed a pilot screen of ~17,000 small molecule compounds with satisfactory performance, and discovered a novel apoE4 modulator that down-regulates apoE4 protein level and inhibits tau phosphorylation in induced pluripotent stem cells (iPSCs)-derived cerebral organoids from AD patient carrying APOE4, demonstrating that our HTS assay is robust and capable of identifying novel apoE4 modulators. Herein, we proposed a collaborative effort to identify apoE4 modulators that down-regulate apoE4 protein level and/or suppress apoE4 aggregation for AD therapy. Aim 1 will complete the apoE4 reporter high-throughput screen on a large chemical library to identify potent and specific apoE4 modulators for suppressing apoE4 level and/or aggregation. Aim 2 will examine the potency, modes of actin (MOA), and therapeutic effects of apoE4 modulators using biochemical and cell-based assays and human iPSC-derived cellular models. Aim 3 will perform SAR-by-catalog and limited chemistry on lead compounds to improve their potency and efficacy and determine the therapeutic efficacy of the most promising candidate compounds in 3-D cerebral organoid models such that our findings may have relevance in a humanized setting. Moreover, the “drug-like” and pharmacokinetics properties and brain penetration will be characterized and benchmarked to position them for future in vivo preclinical animal studies. The identified apoE4 modulators will be promising drug leads in novel therapeutic strategies for AD.

项目概要 阿尔茨海默氏病 (AD) 是伴有认知障碍的最常见痴呆症 β 淀粉样蛋白 (Aβ) 斑块和含有 tau 蛋白的神经原纤维缠结的缺陷和病理性积累。 大多数 AD 病例发生在晚年，通常在 65 岁之后发病，遗传风险最强。 晚发性 AD (LOAD) 的因素是载脂蛋白 E (APOE) 基因型，ε4 等位基因会增加 AD 风险， 与常见的 ε3 等位基因相比，ε2 等位基因具有保护作用。 调节 apoE4 蛋白水平和/或抑制 apoE4 聚集不仅可以缓解淀粉样蛋白病理，而且 还可以防止 tau 介导的神经变性。在我们的初步研究中，我们开发了 apoE4。 具有分裂荧光素酶蛋白片段互补的报告基因检测，使我们不仅能够监测 apoE4 蛋白质水平，还可以通过荧光素酶生物发光信号测量 apoE4 自身寡聚化/聚集 在 Mayo-SBP (Sanford Burnham Prebys) 药物的支持下。 在发现合作计划中，我们对约 17,000 种小分子化合物进行了试点筛选 令人满意的性能，并发现了一种新型 apoE4 调节剂，可下调 apoE4 蛋白水平， 抑制 AD 患者诱导多能干细胞 (iPSC) 来源的脑类器官中 tau 磷酸化 携带 APOE4，证明我们的 HTS 测定是稳健的并且能够识别新型 apoE4 调节剂。 在此，我们提出了一项合作努力，以确定下调 apoE4 蛋白水平的 apoE4 调节剂 和/或抑制 apoE4 聚集用于 AD 治疗目标 1 将完成 apoE4 报告基因高通量。 在大型化学库中进行筛选，以鉴定用于抑制 apoE4 水平的有效且特异性的 apoE4 调节剂 目标 2 将检查 apoE4 的效力、肌动蛋白 (MOA) 模式和治疗效果。 使用生化和基于细胞的测定以及人类 iPSC 衍生的细胞模型的调节剂 Aim 3 将。 对先导化合物进行按目录的 SAR 和有限的化学分析，以提高其效力和功效， 确定 3-D 大脑类器官模型中最有前途的候选化合物的治疗效果 此外，我们的研究结果可能与人性化环境相关。 药代动力学特性和大脑渗透性将被表征和基准测试，以定位它们 未来的体内临床前动物研究中，已确定的 apoE4 调节剂将成为新型药物先导药物。 AD 的治疗策略。