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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）是一种由24个氨基酸组成的脑源性肽，来自于死后散发的 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- 毒性[2]。据报道，内源性HN血浆水平随年龄增长而降低[3]， HN水平的降低与衰老过程中的认知能力下降有关。SNP（rs 2854128）的鉴定使用来自大样本老年人的线粒体基因组HN编码区的线粒体GWAS，成年人进一步支持HN及其循环水平的作用，加速认知能力下降[4]。的高通量筛选（HTS）可格式化测定的可用性，允许筛选以鉴定小分子能够增强AD模型中HN的表达及其水平。在父母补助金中，我们完成了对小分子作为humanin（HN）模拟物，并已确定有效的命中，增加了p- 神经元细胞中的gp 130和p-Akt水平。对这些模拟物的进一步模拟、测试和评估正在进行中遵循拨款中列出的路径。评估HN肽本身水平的新测定法的可用性，在体外和组织中的研究也提供了鉴定提高HN水平的小分子的机会。我们使用该测定法的初步数据显示我们可以检测SH-SY 5 Y细胞中的HN水平。因此对于补充研究，我们建议进行筛选HN增强剂使用人类神经母细胞瘤表达和分泌HN的SH-SY 5 Y细胞[5]。我们在补编中建议的工作是针对对HTS的可用HN ELISA测定进行格式化，并筛选UCLA化合物文库的子集，鉴定改变细胞中HN水平的化合物。筛选中的命中将被验证，二次测试以确认SH-SY 5 Y和iPSC衍生的人神经元中的HN增强，以及小鼠神经母细胞瘤N2 a。还将针对增加的神经元p-Akt评估验证的命中。因此在补充，我们建议扩大父母补助金的目标1，以包括HN增强剂的鉴定沿着我们目前鉴定的HN模拟物将进行进一步的测试和评估，如家长补助金然后，筛选的命中将进行二次测试，以确认HN增强， SH-SY 5 Y和iPSC衍生的人神经元，以及鼠神经母细胞瘤N2 a细胞中。经验证的命中率将也可以使用我们的p-Akt AlphaLISA进行评估，以确认神经元中的生物活性。经验证的HN增强剂将通过体外ADME-T评估，作为母基金Aim 3扩展的一部分进行优先排序测定。优先的类似物将进行药代动力学（PK）和药效学（PD）研究，以鉴定脑渗透性HN增强剂。从补充补助金中鉴定的这种HN增强剂将具有有可能被开发为阿尔茨海默病（AD）的新的治疗候选人，并可能在未来也可以在其他疾病如糖尿病和心血管疾病中进行评估。