Small molecule mimetics of Humanin that normalize neuronal p-Akt as novel therapeutics for AD

护脑素小分子模拟物可使神经元 p-Akt 正常化，作为 AD 的新型疗法

基本信息

批准号：
10810521
负责人：
Varghese John
金额：
$ 27.48万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10810521
关键词：
Acceleration Administrative Supplement Age Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease patient Alzheimer&apos s disease therapeutic Amino Acids Amyloid beta-Protein Animals Area Under Curve Autopsy Award Binding Proteins Biological Biological Assay Brain Cardiovascular Diseases Cells Characteristics Code Data Diabetes Mellitus Disease Dose Drug Kinetics Elderly Enhancers Enzyme-Linked Immunosorbent Assay Euthanasia Evaluation Future Gender Grant Half-Life Hour Human IL6ST gene Impaired cognition In Vitro Left Libraries Link Methods Microsomes Mitochondria Mus N-Methylaspartate Neuroblastoma Neurons Noise Occipital lobe Oral Peptides Permeability Pharmaceutical Preparations Pharmacodynamics Plasma Plasma Proteins Play Property Reporting Research Role Route Sampling Siblings Signal Transduction Testing Time Tissues Toxic effect Validation Work analog brain tissue candidate selection dosage evaluation/testing experimental study genome wide association study high throughput screening humanin in vivo induced pluripotent stem cell mimetics mitochondrial genome mouse model neuroblastoma cell neuroprotection novel therapeutics parent grant pharmacokinetics and pharmacodynamics scale up screening small molecule subcutaneous therapeutic candidate

项目摘要

ABSTRACT (Supplement) Humanin (HN), a 24-amino acid mitochondrial-derived peptide from the occipital lobe of postmortem sporadic AD patient brain tissue [1] is a neuroprotective peptide that could protect neurons from Aβ-related and NMDA- induced toxicity [2]. A decrease in the endogenous HN plasma level with age has been reported [3], and decreased HN levels has been linked to cognitive decline during aging. The identification of a SNP (rs2854128) using mitochondrial GWAS in the HN-coding region of the mitochondrial genome from a large sample of older adults further supports the role of HN and its circulating levels with accelerated cognitive decline [4]. The availability of high throughput screening (HTS) formattable assays, allow for screening to identify small molecules that can enhance HN expression and its levels in AD models. In the parent grant we completed screening for small molecules that act as humanin (HN) mimetics and have identified validated hits that increase both the p- gp130 and p-Akt levels in neuronal cells. Further analoging, testing and evaluation of these mimetics are ongoing following the path outlined in the grant. The availability of new assays to assess levels of HN peptide itself in vitro and in tissue has presented the opportunity to also identify small molecules that enhance HN levels. Our preliminary data using this assay shows that we can detect HN levels in SH-SY5Y cells. Hence for the Supplement research we propose is to conduct screening for HN enhancers using the human neuroblastoma SH-SY5Y cells that express and secrete HN [5]. The work we propose in the Supplement is directed toward formatting of the available HN ELISA assay for HTS and screening of a subset of the UCLA compound library to identify compounds that change HN levels in cells. Hits from the screening would be validated and then undergo secondary testing to confirm HN enhancement in SH-SY5Y and iPSC-derived human neurons, as well as in murine neuroblastoma N2a. Validated hits would also be assessed for increasing neuronal p-Akt. Thus, in the supplement we propose an Expansion of Aim1 of the parent grant to include identification of HN enhancers that along with our currently identified HN mimetics would undergo further testing and evaluation as outlined in the parent grant. Hits from the screening would then undergo secondary testing to confirm HN enhancement in SH-SY5Y and iPSC-derived human neurons, as well as in murine neuroblastoma N2a cells. Validated hits would also be assessed using our p-Akt AlphaLISA to confirm biological activity in neurons. Validated HN enhancers would undergo prioritization as part of Expansion of Aim3 of the parent grant by evaluation in in vitro ADME-T assays. Prioritized analogs would undergo pharmacokinetic (PK) and pharmacodynamics (PD) studies to identify brain permeable HN enhancers. Such an HN enhancers identified from the supplement grant would have the potential to be developed as novel therapeutic candidates for Alzheimer's disease (AD), and could in the future also be evaluated in other disorders such as diabetes and cardiovascular disease.

摘要（补充） Humanin（Hn），一种24-氨基酸线粒体衍生的肽，来自枕孢子的枕叶 AD患者脑组织[1]是一种神经保护肽，可以保护神经元免受Aβ相关和NMDA- 诱导毒性[2]。随着年龄的报道，内源性HN血浆水平降低[3]，并且开发的HN水平与衰老期间的认知下降有关。识别SNP（RS2854128）使用线粒体基因组的HN编码区域中使用线粒体GWA，从大量较老的样本中成年人进一步支持HN及其循环水平的作用，并加速认知下降[4]。这可用性高通量筛选（HTS）格式测定，允许筛选识别小分子这可以增强HN表达及其在AD模型中的水平。在父母赠款中，我们完成了筛选充当人类（HN）模拟物并鉴定出验证的命中的小分子，这两者都会增加P- 神经元细胞中的GP130和P-AKT水平。这些模拟物的进一步模拟，测试和评估正在进行中遵循赠款中概述的道路。新的评估可用于评估HN胡椒本身的水平体外和组织中已经提供了鉴定提高HN水平的小分子的机会。我们的使用此测定的初步数据表明，我们可以检测SH-SY5Y细胞中的HN水平。因此我们建议的补充研究是使用人类神经母细胞瘤对HN增强子进行筛查表达和秘密HN的SH-SY5Y细胞[5]。我们在补品中提出的工作是针对的用于HTS的可用HN ELISA分析的格式化以及将UCLA复合库的子集筛选到识别改变细胞中HN水平的化合物。筛选中的命中将得到验证，然后进行辅助测试以确认SH-SY5Y和IPSC衍生的人类神经元的HN增强以及鼠神经母细胞瘤N2A。还将评估经过验证的命中，以增加神经元P-AKT。那在补充我们建议扩展父母赠款的AIM1，以包括识别HN增强剂与我们当前确定的HN Mimetics一起，将进行进一步的测试和评估父母赠款。然后，筛选的命中将进行辅助测试，以确认HN增强 SH-SY5Y和IPSC衍生的人神经元以及鼠神经母细胞瘤N2A细胞。经过验证的命中会也可以使用我们的p-akt alphalisa评估以确认神经元中的生物学活性。经过验证的HN增强剂通过评估体外ADME-T，将作为父母赠款AIM3的扩展的一部分进行优先级排序测定。优先的类似物将接受药代动力学（PK）和药效学（PD）研究，以鉴定大脑可渗透的HN增强剂。从补充赠款中确定的这样的HN增强剂将具有作为阿尔茨海默氏病（AD）的新型治疗候选者的潜力，将来可能还可以在其他疾病中进行评估，例如糖尿病和心血管疾病。