Evaluating the p-Tau inhibition and neuroprotective effects of sAPPalpha using brain permeable small molecules

使用脑通透性小分子评估 sAPPalpha 的 p-Tau 抑制和神经保护作用

• 批准号: 10522638
• 负责人: Varghese John
• 金额: $ 39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522638
• 关键词: 3xTg-AD mouse; ADME Study; Acute; Aducanumab; Affect; Affinity; Agonist; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease therapy; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Antibodies; Artificial Membranes; Behavioral; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Availability; Brain; Cause of Death; Cell Membrane Permeability; Cells; Chemicals; Chemistry; Chronic; Cognition; Cognitive; Corticotropin-Releasing Hormone; Country; Data; Development; Dose; Drug Kinetics; Enhancers; Evaluation; Extracellular Domain; FDA approved; Goals; HTR3A gene; Hand; Hippocampus (Brain); Histopathology; Human; Impaired cognition; In Vitro; Induced pluripotent stem cell derived neurons; Injections; J20 mouse; Ligands; Membrane; Memory; Mus; Muscarinic M1 Receptor; Muscarinic M3 Receptor; Neurites; Neurofibrillary Tangles; Neurons; Oral; Outcome; Pathology; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phosphorylation Site; Phosphotransferases; Postoperative Nausea and Vomiting; Property; Reporting; Running; Safety; Serotonin Antagonists; Site; Solubility; Structure-Activity Relationship; Synapses; Tauopathies; Testing; Tropisetron; Vertebral column; acetylcholine receptor agonist; aged; alpha-bungarotoxin receptor; analog; antagonist; apolipoprotein E-4; base; biological adaptation to stress; cognitive benefits; combinatorial; density; design; efficacy study; improved; in vivo; in vivo evaluation; induced pluripotent stem cell; lead candidate; mouse model; neuroblastoma cell; neurofibrillary tangle formation; neuroprotection; new therapeutic target; novel therapeutic intervention; novel therapeutics; phosphoproteomics; presenilin-1; receptor; receptor binding; response; screening; serotonin receptor; small molecule; tau Proteins; tau aggregation; tau phosphorylation; tau-1; therapeutic candidate; therapeutically effective

PROJECT SUMMARY/ABSTRACT We previously reported (2) on the soluble amyloid precursor protein alpha (sAPPα)-enhancing effects of tropisetron (F03). Highly brain-permeable F03 is approved in 49 countries for the treatment of post-operative nausea and vomiting (PONV) and is a multifunctional ligand: it is a potent 5-HT3 serotonin receptor (5-HT3R) antagonist (Ki ~ 3 nM), a partial α7 nicotinic acetylcholine receptor (α7nAChR) agonist (Ki ~ 450 nM), and binds to the extracellular domain of amyloid precursor protein (eAPP). In our studies, we found F03 increases sAPPα and also significantly decreases the phospho-tau (p-tau)/total tau (t-tau) ratio in two Alzheimer's disease (AD) mouse models. We show that F03 can also reduce corticotropin-releasing factor (CRF) induced p-tau/tau increase in vitro. Hyperphosphorylation of tau leads to toxic tau oligomers and ultimately formation of neurofibrillary tangles (NFTs) in AD brain and is closely correlated with cognitive decline (34); thus, decreasing tau phosphorylation is a critical target for new therapeutic approaches for AD and tauopathies. Based on reports that sAPPα reduces the p-tau/t-tau ratio through suppression of activity of the kinase GSK3β (9), we plan to assess the ability of our sAPPα enhancers to reduce the p-tau/t-tau ratio in vitro and in vivo. A key goal of the project is to identify optimized small molecule sAPPα-enhancing, p-tau/t-tau lowering compounds that improve cognition in murine AD models, for further development as a novel therapy for AD. An additional goal would be to elucidate the underlying mechanism of action (MOA) of sAPPα enhancement and related reduction in the p- tau/t-tau ratio. In Aim 1, we would test F03 and analogs we have in-hand as well as new analogs from Aim 2 in SH-SY5Y cells and 3xTg-AD primary neurons to establish EC50s for sAPPα enhancement and p-tau/t-tau decreases. Prioritized compounds would be evaluated for receptor binding and APP binding. Tertiary testing will be in induced pluripotent stem cell (iPSC)-derived neurons from AD subjects and includes synaptic spine density quantification. In Aim 2, medicinal chemistry would be used to design new chemical entity (NCE) analogs in a iterative fashion with receptor and APP binding along with enhanced drug-like properties and oral brain permeability. In Aim 3, ADME studies on analogs include solubility, microsomal stability, protein binding, parallel artificial membrane permeability assay (PAMPA) analysis, and in vivo pharmacokinetics (PK). Optimal candidates would undergo safety (including hERG) and off-target panel profiling along with analyses of effect on other kinases and of tau phosphorylation sites. In Aim 4 in vivo testing, including acute studies on select candidates to evaluate effect on intracerebroventricular (ICV) delivered CRF induced p-tau/t-tau increases in brain in 3xTg-AD mice. Active candidates would be tested in chronic 4-week efficacy studies in 3xTg-AD and ApoE4-5XFAD AD model mice. Readouts would include behavioral and histopathology analysis, including hippocampal neurite load and neuronal spine density. Biochemical outcomes include p-tau/t-tau ratio, sAPPα and Aβ. Mechanistic studies by phosphoproteomics used for correlations between readouts and cognition.

项目总结/文摘