Alzheimer's disease pYGSK3 pathophysiology and PTPRD positive allosteric modulators

阿尔茨海默病 pYGSK3 病理生理学和 PTPRD 正变构调节剂

• 批准号: 10286886
• 负责人: George Richard Uhl
• 金额: $ 34.94万
• 依托单位: BIOMEDICAL RESEARCH INSTITUTE OF NEW MEX
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10286886
• 关键词: 3xTg-AD mouse; Aging; Alzheimer disease prevention; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Apolipoprotein E; Behavioral; Binding; Binding Sites; Biodistribution; Biological Availability; Brain; Catalytic Domain; Clinical; Data; Defect; Dementia; Development; Dietary intake; Docking; Dose; Early treatment; Enzymes; Flavanol; Flavones; Functional disorder; Funding; Genes; Genetic; Glycogen Synthase Kinases; Histologic; In Vitro; Incidence; Intake; Lead; Modeling; Mus; Neurofibrillary Tangles; Neurons; Pathogenicity; Pathologic; Pathology; Pathway interactions; Phosphopeptides; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Pilot Projects; Property; Protein Dephosphorylation; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Quercetin; Reagent; Reporting; Role; Serine; Site; Specificity; Structure; Structure-Activity Relationship; Study models; Test Result; Testing; Threonine; Time; Toxic effect; Translations; Tyrosine; Tyrosine Phosphorylation; Validation; Variant; Work; acute toxicity; age related; analog; density; dietary; drug candidate; drug market; feasibility testing; genetic association; hyperphosphorylated tau; improved; in silico; in vivo; in vivo evaluation; innovation; insight; neuropathology; new therapeutic target; novel; novel strategies; off-target site; phosphoproteomics; positive allosteric modulator; prevent; receptor; tau Proteins; tau aggregation; tau-1; therapy development

Project Summary/Abstract Alzheimer's disease (AD) receives pathogenic contributions from genetics [1, 2] and from environmental influences that include dietary intake of flavanols > flavones [4-8]. Neurofibrillary tangles (NFTs) rich in hyperphosphorylated tau protein [10] are prominent features of AD neuropathology. NFT densities correlate well with the degree of AD dementia [15] [18] and are influenced by variation in the ApoE and PTPRD genes [3]. One approach to altering tau/NFT pathophysiology is to reduce activities of the kinases that hyper- phosphorylate tau. The glycogen synthase kinases GSK3α and GSK3β are prominent tau phosphorylators [19]. GSK3α and GSK3β are activated by phosphorylation of their own tyrosines (pY279 and pY216) by known tyrosine kinases [20] [21, 22]. Increasing activity of tyrosine phosphatase(s) that dephosphorylate and reduce activities of brain GSK3α and GSK3β thus provides a novel approach to reducing tau pathology in AD. Evidence (much developed with support from our first NIA supplement) now supports roles for: a) the receptor type protein tyrosine phosphatase PTPRD as both a i) key physiological phosphatase for phospho (pY) GSK3α and GSK3β and ii) novel target for decreasing pathological AD tau hyper- phosphorylation and b) flavanols as lead compound PTPRD positive allosteric modulators (PAMs) that increase this desired PTPRD activity. We will enhance this evidence and move toward translation by testing hypotheses that a) increased PTPRD dephosphorylation of GSK3β and GSK3α reduces the activities of these tau-hyperphosphorylating kinases with specificity, underlying PTPRD's genetic associations with NFT densities in AD brains and b) flavanols whose intake reduces AD incidence in aging [4-8] serve as PAMs for PTPRD's phosphatase, increase GSK3 dephosphorylation with specificity and provide a pathway for development of improved, specific PTPRD PAMs that can reduce progression to AD deficits during aging. We will test these hypothesis and support development and translation of PTPRD PAMs in several ways: 1) we will characterize the specificity of PTPRD effects by comparing i: rates of PTPRD dephosphorylation of pYGSK3α and pYGSK3β and ii: quercetin effects on these rates vs those for each of > 80 candidate neuronal PTPRD substrates. 3) We will synthesize and test novel flavanol analogs as improved PTPRD PAMs, nominating novel structures by in silico docking to the PAM vs catalytic sites on PTPRD's phosphatase, testing these structures in vitro, refining our in silico models and nominating/synthesizing/testing new structures on the basis of these results. For the best candidate positive allosteric modulators, we will test specificities vs other PTPRD substrate phosphopeptides and off-target sites of action of currently marketed drugs. We will test the most promising PTPRD PAMs in vivo for gross, histological or behavioral toxicities, biodistribution (including brain) and target engagement. We will expand validation of quercetin effects in aging 3xTg- AD mice and initiate studies to test novel PTPRD PAM(s) in this model. This work will advance our understanding of AD pathophysiology, validate novel approaches to PTPRD positive allosteric modulation and provide a basis for development of interventions that can prevent and/or treat key aspects of AD pathophysiology.

项目总结/摘要 阿尔茨海默病（AD）接受遗传学[1，2]和 环境影响，包括黄烷醇>黄酮的饮食摄入[4-8]。 富含过度磷酸化tau蛋白的神经纤维缠结（NFT）[10]是突出的 AD神经病理学特征。NFT密度与AD痴呆的程度密切相关 [15][18]并受ApoE和PTPRD基因变异的影响[3]。 改变tau/NFT病理生理学的一种方法是降低过度激活tau/NFT的激酶的活性。 磷酸化tau。糖原合成酶激酶GSK 3 α和GSK 3 β是主要的tau蛋白， 磷酸化酶[19]。GSK 3 α和GSK 3 β通过其自身酪氨酸的磷酸化而活化 （pY 279和pY 216）通过已知的酪氨酸激酶[20] [21，22]。增加酪氨酸活性 磷酸酶，使脑GSK 3 α和GSK 3 β脱磷酸化并降低活性， 提供了一种减少AD中tau病理的新方法。 证据（在我们的第一个NIA补充的支持下得到了很大的发展）现在支持以下角色：a） 受体型蛋白酪氨酸磷酸酶PTPRD作为i）关键生理磷酸酶 用于磷酸化（pY）GSK 3 α和GSK 3 β，以及ii）用于降低病理性AD tau蛋白超表达的新靶点。 磷酸化和B）黄烷醇作为先导化合物PTPRD正变构调节剂（PAM） 增加所需的PTPRD活性。我们将加强这一证据并走向 通过测试以下假设进行翻译：a）增加GSK 3 β的PTPRD去磷酸化， GSK 3 α特异性地降低这些tau超磷酸化激酶的活性，这是潜在的 PTPRD与AD脑中NFT密度的遗传相关性和B）黄烷醇的摄入 降低老年AD发病率[4-8]，作为PTPRD磷酸酶的PAM，增加GSK 3 特异性去磷酸化，并为开发改善的，特异性的 PTPRD PAM可以减少衰老过程中AD缺陷的进展。 我们将测试这些假设，并支持PTPRD PAM的开发和翻译， 方法：1）我们将通过比较i：PTPRD的发生率来表征PTPRD效应的特异性 pYGSK 3 α和pYGSK 3 β的去磷酸化和ii：槲皮素对这些速率的影响与 每种> 80种候选神经元PTPRD底物。3)我们将合成并测试新型黄烷醇 类似物作为改进的PTPRD PAM，通过与PAM的计算机对接提名新结构， PTPRD磷酸酶的催化位点，体外测试这些结构， 模型和提名/合成/测试新的结构的基础上，这些结果。为 最佳候选正变构调节剂，我们将测试与其他PTPRD底物的特异性 磷酸肽和目前市售药物的脱靶作用位点。我们将测试最 有希望的体内PTPRD PAM的大体、组织学或行为毒性、生物分布 （包括大脑）和目标参与。我们将扩大验证槲皮素在衰老中的作用 3xTg-AD小鼠，并启动研究以在该模型中测试新型PTPRD PAM。这项工作将 推进我们对AD病理生理学的理解，验证PTPRD阳性的新方法 变构调节，并为开发可以预防和/或 治疗AD病理生理学的关键方面。

项目成果

Structure-activity studies of PTPRD phosphatase inhibitors identify a 7-cyclopentymethoxy illudalic acid analog candidate for development.

• DOI: 10.1016/j.bcp.2021.114868
• 发表时间: 2022-01
• 期刊: BIOCHEMICAL PHARMACOLOGY
• 影响因子: 5.8
• 作者: Henderson, Ian M.;Zeng, Fanxun;Bhuiyan, Nazmul H.;Luo, Dan;Martinez, Maria;Smoake, Jane;Bi, Fangchao;Perera, Chamani;Johnson, David;Prisinzano, Thomas E.;Wang, Wei;Uhl, George R.
• 通讯作者: Uhl, George R.

Substrate-selective positive allosteric modulation of PTPRD's phosphatase by flavonols.

• DOI: 10.1016/j.bcp.2022.115109
• 发表时间: 2022-08
• 期刊: BIOCHEMICAL PHARMACOLOGY
• 影响因子: 5.8
• 作者: Henderson, Ian M.;Marez, Carlissa;Dokladny, Karol;Smoake, Jane;Martinez, Maria;Johnson, David;Uhl, George R.
• 通讯作者: Uhl, George R.