Novel small molecules targeting brain phospholipid dysregulation in AD

针对 AD 脑磷脂失调的新型小分子

• 批准号: 10457810
• 负责人: Dongming Cai
• 金额: --
• 依托单位: JAMES J PETERS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10457810
• 关键词: AD transgenic mice; APP-PS1; Abeta clearance; Aging; Alzheimer' s Disease; Alzheimer' s disease therapy; American; Amyloid; Amyloid beta-42; Amyloid beta-Protein; Animals; Apolipoprotein E; Astrocytes; Attenuated; Behavioral; Biological Assay; Biological Availability; Brain; Calcium Channel; Calcium Channel Inhibition; Candidate Disease Gene; Chronic; Cognitive; Cognitive deficits; Confocal Microscopy; Cycloheximide; Data; Data Set; Dementia; Development; Diagnosis; Dissection; Dose; Down-Regulation; Drug Delivery Systems; Drug usage; Enzymes; FDA approved; Female; Funding; Generations; Genes; Grant; Healthcare Systems; Hippocampus (Brain); Immunofluorescence Immunologic; Impaired cognition; In Vitro; Intravenous; Investigation; Knock-in Mouse; Lead; Libraries; Link; Luciferases; Maps; Measures; Mediating; Medical; Modification; Morbidity - disease rate; Mus; Nature; Neurodegenerative Disorders; Neurons; Nimodipine; Oral; Oral Administration; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Penetrance; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phosphatidylinositol 4,5-Diphosphate; Phospholipids; Phosphoric Monoester Hydrolases; Physiologic pulse; Pilot Projects; Post-Traumatic Stress Disorders; Prevalence; Proteins; Proto-Oncogene Proteins c-akt; Research; Role; Route; SYNJ1 gene; Services; Signal Transduction; Synapses; System; Testing; Therapeutic; Transgenic Organisms; Traumatic Brain Injury; United States Department of Veterans Affairs; Veterans; Veterans Health Administration; analog; apolipoprotein E-3; apolipoprotein E-4; base; cognitive function; design; efficacious treatment; gene network; genetic manipulation; improved; in vitro testing; in vivo; in vivo Model; insight; intraperitoneal; male; mild traumatic brain injury; mortality; mouse model; neuropathology; novel; novel therapeutics; overexpression; pharmacokinetics and pharmacodynamics; preclinical efficacy; research study; screening; side effect; small molecule; tau Proteins; tau-1; time interval; transcriptome sequencing; treatment strategy; uptake; AD transgenic mice; APP-PS1; Abeta clearance; Aging; Alzheimer' s Disease; Alzheimer' s disease therapy; American; Amyloid; Amyloid beta-42; Amyloid beta-Protein; Animals; Apolipoprotein E; Astrocytes; Attenuated; Behavioral; Biological Assay; Biological Availability; Brain; Calcium Channel; Calcium Channel Inhibition; Candidate Disease Gene; Chronic; Cognitive; Cognitive deficits; Confocal Microscopy; Cycloheximide; Data; Data Set; Dementia; Development; Diagnosis; Dissection; Dose; Down-Regulation; Drug Delivery Systems; Drug usage; Enzymes; FDA approved; Female; Funding; Generations; Genes; Grant; Healthcare Systems; Hippocampus (Brain); Immunofluorescence Immunologic; Impaired cognition; In Vitro; Intravenous; Investigation; Knock-in Mouse; Lead; Libraries; Link; Luciferases; Maps; Measures; Mediating; Medical; Modification; Morbidity - disease rate; Mus; Nature; Neurodegenerative Disorders; Neurons; Nimodipine; Oral; Oral Administration; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pathway interactions; Penetrance; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phosphatidylinositol 4,5-Diphosphate; Phospholipids; Phosphoric Monoester Hydrolases; Physiologic pulse; Pilot Projects; Post-Traumatic Stress Disorders; Prevalence; Proteins; Proto-Oncogene Proteins c-akt; Research; Role; Route; SYNJ1 gene; Services; Signal Transduction; Synapses; System; Testing; Therapeutic; Transgenic Organisms; Traumatic Brain Injury; United States Department of Veterans Affairs; Veterans; Veterans Health Administration; analog; apolipoprotein E-3; apolipoprotein E-4; base; cognitive function; design; efficacious treatment; gene network; genetic manipulation; improved; in vitro testing; in vivo; in vivo Model; insight; intraperitoneal; male; mild traumatic brain injury; mortality; mouse model; neuropathology; novel; novel therapeutics; overexpression; pharmacokinetics and pharmacodynamics; preclinical efficacy; research study; screening; side effect; small molecule; tau Proteins; tau-1; time interval; transcriptome sequencing; treatment strategy; uptake

Alzheimer’s disease (AD) is the most prevalent neurodegenerative disease of aging, with one in eight older Americans diagnosed with AD. It is also the most frequently diagnosed type of dementia within the Veterans Affairs (VA) Medical System, and one of the major causes of morbidity and mortality among veterans. The Department of VA estimates that 600,000 veterans suffered from severe AD and other forms of dementia in 2000, and this number is increased significantly today, because of the increasing proportion of older veterans and the increased prevalence of dementia in veterans that suffer from traumatic brain injury (TBI) and/or post- traumatic stress disorder (PTSD). Therefore, AD research studies are particularly important to veterans. Currently no treatment is available to slow or stop AD. There is a great need for identification of more efficacious therapies for AD, which is among the priorities of VA RR&D research directions. Synaptojanin 1 (synj1), the main phosphoinositol biphosphate phosphatase [PIP2 degrading enzyme] in the brain and synapses, has been recently linked to AD. More importantly, we have demonstrated that synj1 regulates lysosomal clearance of Aβ and that the increased synj1 expression links to ApoE4-induced phospholipid dysregulation and cognitive deficits. Down-regulation of synj1 promotes Aβ clearance, reduces tau hyper-phosphorylation and ameliorates ApoE4 pathogenic effects. Subsequently, reduction of synj1 attenuates AD-related pathological changes and behavioral deficits in AD mouse models. These findings suggest that reduction of synj1 has potential therapeutic benefits for AD. Our initial screening of 89 top hits out of a library of compounds (~3,600 small molecules) with the potential to reduce synj1 protein levels using “The Connectivity Map”, identified a FDA-approved drug nimodpine with synj1- and Aβ-lowering effects in both wild- type and ApoE4 neuronal cultures. Further administration of nimodipine for one month, is capable of reducing brain content of synj1 and Aβ, as well as improving cognitive functions in an AD transgenic mouse model and an ApoE4 KI mouse model. However, chronic administration of nimodipine failed to reduce brain Aβ42 levels (particularly insoluble fractions), or to improve cognitive function. Our data suggest that the effects of nimodipine on reduction of synj1 expression are independent of its inhibitory effects on calcium channel activities. We then designed and synthesized first-generation nimodipine structural analogs using medicinal chemistry to reduce its calcium channel activity, and identified a novel compound, SynaptoCpd#9, with attenuated inhibition of calcium channels and increased potency against synj1 and Aβ42 compared to nimodipine both in vitro and in vivo. Oral administration of SynaptoCpd#9 in APPSwe/PS1E9 and ApoE4 mice for 3-6 months improved cognitive function and reduced AD-related pathologies (insoluble Aβ42 particularly). RNA-sequencing and qPCR studies of treated ApoE4 neurons identified three candidate genes involved in nimodipine- or SynaptoCpd#9-mediated effects. We posit that continued exploration of SAR of nimodipine derivatives and further dissection of their mechanisms of actions will provide new insights regarding AD pathogenesis and may lead to identification of novel targets for AD therapies. In this application, we propose to refine structural modifications of nimodipine derivatives to increase their potency at lowering synj1 and improving cognitive function in AD mouse models (aim 1: medicinal chemistry modifications, in vitro test funnel assays and in vivo proof of concept animal studies). In parallel studies, we propose to dissect mechanisms of actions of these novel compounds (aim 2: mechanistic studies), which will provide new insights regarding mechanisms of AD pathogenesis and advance therapeutic strategies for AD. Information obtained from our proposed studies will lead to development of novel potent AD therapies, and better understanding of AD pathogenesis through mechanistic investigation. Therefore, our studies will directly benefit Veterans, and improve the quality of service provided within the VA health care system.

阿尔茨海默氏病（AD）是衰老的最普遍的神经退行性疾病 美国人被诊断出患有广告。它也是退伍军人中最常见的痴呆症类型 事务（VA）医疗系统，也是退伍军人发病和死亡的主要原因之一。 弗吉尼亚州估计，有60万退伍军人遭受了严重的广告和其他形式的痴呆 2000年，由于老年退伍军人比例的增加，今天这个数字显着增加 以及遭受脑外伤（TBI）和/或后遭受创伤性脑损伤（TBI）的退伍军人痴呆症患病率的增加 创伤应力障碍（PTSD）。因此，AD研究对退伍军人尤为重要。 目前尚无治疗方法可以减速或停止广告。非常需要识别更多 AD的有效疗法，这是VA RR＆D研究方向的优先级。 Synaptojanin 1（Synj1），主要磷酸肌醇双磷酸磷酸酶[PIP2降解酶] 大脑和突触最近已与AD联系在一起。更重要的是，我们已经证明了Synj1 调节Aβ的溶酶体清除率，并增加Synj1表达与APOE4诱导的联系 磷脂失调和认知缺陷。 Synj1的下调促进Aβ清除率，减少 Tau高磷酸化并改善APOE4致病作用。随后，Synj1的减少 减弱与广告相关的病理变化，行为在AD小鼠模型中定义。这些发现 表明Synj1的减少对AD具有潜在的治疗益处。我们最初筛选了89个热门单曲 一个化合物库（约3,600个小分子），具有使用“使用”的潜力 连通图”，确定了由FDA批准的药物Nimodpine，在这两个野生中都具有synj1-和降低Aβ的作用 类型和APOE4神经元文化。进一步给予氮氨胺一个月，能够降低 Synj1和Aβ的大脑含量，以及在AD转基因小鼠模型中改善认知功能 APOE4 Ki鼠标模型。但是，慢性氮氨基氨酸的施用未能降低大脑Aβ42水平 （部分不溶性分数）或改善认知功能。我们的数据表明 尼莫地平减少synj1表达与其对钙通道的抑制作用无关 活动。然后，我们使用医学设计和合成第一代氮瞬平结构类似物 化学以减少其钙通道活性，并确定一种新型化合物SynaptOCPD＃9， 与Synj1和Aβ42的钙通道的抑制减弱，与 尼莫迪平均体外和体内。 Appswe/ps1e9和apoE4小鼠的synaptoCPD＃9的口服给药 在3-6个月中，改善了认知功能和降低与AD相关的病理（尤其是不溶于Aβ42）。 对APOE4神经元进行的RNA测序和QPCR研究确定了涉及的三个候选基因 nimodipine-或SynaptoCPD＃9介导的效果。我们指出，继续探索nimodipine的SAR 衍生物和进一步解剖其行动机制将为广告提供新的见解 发病机理，可能导致对AD疗法的新靶标识别。 在此应用中，我们建议完善对氮岛衍生物的结构修饰，以增加其 降低Synj1并改善AD小鼠模型的认知功能方面的效力（AIM 1：药物化学 修改，体外测试漏斗分析和体内概念动物研究证明）。在平行研究中，我们 提出剖析这些新型化合物作用机制的提议（AIM 2：机械研究），这将 提供有关AD发病机理的机制和AD提前理论策略的新见解。 从我们提出的研究中获得的信息将导致开发新颖的有效AD疗法，并 通过机械研究更好地了解AD发病机理。因此，我们的研究将直接 受益于退伍军人，并提高VA医疗保健系统中提供的服务质量。