Novel Antagonists of the N-terminal Domain of the CRF Receptor Type 1 for Alzheimer's Disease

治疗阿尔茨海默病的 1 型 CRF 受体 N 末端结构域的新型拮抗剂

• 批准号: 10433769
• 负责人: Robert A Rissman
• 金额: $ 35.45万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10433769
• 关键词: AD transgenic mice; Academia; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Amyloid beta-Protein; Animal Model; Animals; Area; Award; Benchmarking; Binding; Biological Assay; Biotin; Brain; C-terminal; CRF receptor type 1; Cells; Chemicals; Clinical Trials; Collaborations; Corticotropin-Releasing Hormone Receptors; Critical Pathways; Development; Disease Progression; Extracellular Domain; Fluorescence Resonance Energy Transfer; Freezing; Funding; Generations; Genomics; Hippocampus (Brain); Hormones; Hypothalamic structure; Impaired cognition; In Vitro; Institutes; Label; Lead; Libraries; Ligand Binding; Measures; Mediating; Mediator of activation protein; Medical; Membrane; Mus; N-terminal; Neuropeptides; Parents; Pathology; Penetrance; Pharmaceutical Chemistry; Pharmacologic Substance; Research; Risk Factors; Rodent; Senile Plaques; Signal Transduction; Slice; Specificity; Stress; Structure-Activity Relationship; Testing; Tissues; Toxic effect; Up-Regulation; Validation; Work; antagonist; base; beta amyloid pathology; brain tissue; efficacy study; experimental study; high throughput screening; in vivo; lead optimization; maltose-binding protein; molecular modeling; mouse model; neurochemistry; neuropathology; novel; pharmacokinetics and pharmacodynamics; radioligand; receptor; scaffold; scale up; small molecule; tau Proteins; tau phosphorylation

Project Summary/Abstract CRF is the major neuropeptide hormone associated with stress and is mediated predominantly signaling through CRF receptor subtype 1 (CRFR1) in the CNS. CRF signaling has been implicated in Alzheimer’s disease (AD) as there is increased CRF expression, CRF binding to and upregulation of CRFR1 receptors in the cortex, hippocampus and hypothalamus appears to occur early in disease AD progression and are prominent neurochemical changes in areas vulnerable to AD neuropathology. CRFR1 expression also intersects with Abeta (Aβ) and tau pathologies, both known to be critical hallmarks of AD. There have been substantial efforts in academia and pharmaceutical companies to develop potent small molecule antagonists of CRFR1, however, even the current 3rd generation antagonists are still directed against the C- terminal regions of the CRFR1 receptor are still based on the 2nd generation chemical scaffolds along with their inherent toxicity liabilities. This project capitalizes on the molecular model of CRFR1 activation, established through extensive studies, where the first step is binding of CRF to specific ligand binding motifs in the receptor’s N-terminal region and binding motifs. We have developed and validated an novel homogeneous TR-FRET assay to measure the direct binding of biotin-labeled CRF to the N-terminal extracellular domain (ECD) of maltose binding protein labeled CRFR1 compatible with very high-throughput screening. We have developed this assay, verified that unlabeled CRF displaces biotin-CRF, and that several known C-terminal CRFR1 antagonists do not displace CRF from the N-term CRFR1 ECD. Motivated by these recent experiments, we propose to screen a large library of compounds against N-terminal CRFR1 ECD in collaboration with the Conrad Prebys Center for Chemical Genomics at Sanford Burnham Prebys Medical Discovery Institute. In Aim 1, we will fully implement this primary N- terminal CRFR1 ECD assay and complete an HTS campaign to identify and confirm candidate N-terminal CRFR1 antagonists. Aim 2 will validate these hits for potency, specificity, functional antagonism and elucidate emergent structure activity relationships (SAR) to assess their chemical tractability. Aim 3 will advance validated hits through hit-to-lead through cycles of medicinal chemistry and testing through 2° critical path assays for potency and functional antagonism, then additional lead optimization cycles where additionally, potency and efficacy in biologically relevant 3° assays for target engagement (e.g. brain tissue slices). The best 2-3 probe compound(s), will be scale-up to benchmark their ADME/T, PK/PD and brain penetrance and in vivo proof-of-concept studies in mouse AD models.

项目总结/摘要 CRF是与应激相关的主要神经肽激素，主要通过信号传导介导 通过CNS中的CRF受体亚型1（CRFR 1）。CRF信号与阿尔茨海默氏症有关 由于CRF表达增加，CRF与CRFR 1受体的结合和CRFR 1受体的上调， 皮质、海马和下丘脑似乎在疾病AD进展的早期发生， 在易受AD神经病理学影响的区域出现显著的神经化学变化。CRFR 1表达也 与Abeta（Aβ）和tau病理学交叉，两者都已知是AD的关键标志。有 在学术界和制药公司的大量努力，开发有效的小分子拮抗剂， 然而，即使是目前的第三代拮抗剂仍然针对CRFR 1的C-末端区域 的CRFR 1受体仍然是基于第二代化学支架沿着其固有的毒性 负债该项目利用CRFR 1激活的分子模型，通过广泛的 研究，其中第一步是CRF与受体N-末端的特异性配体结合基序结合 区域和结合基序。我们已经开发并验证了一种新的均相TR-FRET测定， 测量生物素标记的CRF与麦芽糖的N-末端胞外结构域（ECD）的直接结合 结合蛋白标记的CRFR 1与非常高通量的筛选相容。按照本 试验证实，未标记的CRF取代生物素-CRF，并且几种已知的C-末端CRFR 1 拮抗剂不从N-末端CRFR 1 ECD置换CRF。受这些实验的启发，我们 我建议与联合国开发计划署合作，筛选针对N-末端CRFR 1 ECD的大型化合物库， Conrad Prebys化学基因组学中心，Sanford Burnham Prebys医学发现研究所。 在目标1中，我们将全面实施这种初级N-末端CRFR 1 ECD检测，并完成HTS活动 以鉴定和确认候选N-末端CRFR 1拮抗剂。目标2将验证这些命中的效力， 特异性，功能拮抗作用和阐明紧急结构活性关系（SAR），以评估其 化学易处理性目标3将通过医疗周期中的命中率来提高经验证的命中率。 通过2°关键路径试验进行化学和效价和功能拮抗试验，然后进行额外的 此外，在生物学相关3°测定中， 接合（例如，脑组织切片）。最好的2-3种探针化合物将被放大以基准测试其 小鼠AD模型中的ADME/T、PK/PD和脑转移率以及体内概念验证研究。