Targeting Necroptosis for Neurodegenerative Disease Therapy

针对神经退行性疾病治疗的坏死性凋亡

• 批准号: 10253999
• 负责人: May Khanna
• 金额: $ 44.96万
• 依托单位: REGENERIX LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10253999
• 关键词: Active Sites; Affect; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Amyotrophic Lateral Sclerosis; Apoptosis; Binding; Binding Proteins; Binding Sites; Biological Assay; Biological Sciences; Brain; Cell Death; Cell membrane; Cells; Cellular Assay; Chemicals; Cholinesterase Inhibitors; Communicable Diseases; Complex; Crystallization; Dimethyl Sulfoxide; Disease; Docking; Drug Industry; Ensure; Exhibits; Extravasation; FDA approved; Follow-Up Studies; Generations; Goals; Grant; Human; In Vitro; Inflammatory; Ischemia; Laboratories; Laboratory Research; Lead; Measures; Memantine; Membrane; Methods; Molecular; Mutate; Necrosis; Neoplasm Metastasis; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Phenylalanine; Phosphotransferases; Property; Protein Kinase; Proteins; RIPK1 gene; RIPK3 gene; Reperfusion Therapy; Reporting; Risk; Science; Signal Transduction; Site; Structural Protein; Structure; TNF gene; Temperature; Testing; Therapeutic; Triage; Work; analog; base; chemical synthesis; clinical candidate; drug development; efficacy testing; experience; in silico; induced pluripotent stem cell; inhibitor/antagonist; innovation; kinase inhibitor; lead optimization; lead series; liver injury; neuron loss; next generation; novel; novel strategies; protein complex; protein protein interaction; proteostasis; renal damage; research and development; scaffold; side effect; small molecule; small molecule libraries; stem cells; synergism; therapeutic target; tumor

PROJECT SUMMARY/ABSTRACT The long-term objective of the project is to develop therapeutics for Alzheimer’s that inhibit a unique method of cell death known as necroptosis. This cell death pathway has been known to be upregulated in neurodegenerative diseases, highlighting the need to find therapeutics that target the proteins involved. Several kinases are known to be important regulators that include: RIPK1, RIPK3 and MLKL. However, unlike most therapeutic approaches that target the kinase portion of RIPK proteins involved in necroptosis, we are targeting the protein-protein interface between RIPK3 and MLKL. This targeting method should uncover more specific compounds since RIPK kinases are involved in multiple pathways. By targeting an interface that is only shared between two proteins involved in necroptosis (protein-protein interface), we ensure that there are fewer side effects as compared to targeting the kinase function implicated in more than just necroptosis. It was previously shown that an interaction between RIPK3 and MLKL located away from the kinase interface is necessary to promote RIPK3 and MLKL binding. We targeted in silico this pocket that is involved in the interaction of RIPK3 to MLKL using a library of small molecules. The top 13 compounds were tested using flow to assay of cells where apoptosis is inhibited and cells only undergo cell death through necroptosis. Using this assay, we were able to narrow down the list of compounds to two hits that specifically inhibit necroptosis, we showed the compounds are targeting the RIPK3 protein in cells. Follow-up studies with derivatives of these two hit compounds exhibited improvements. We propose to further develop compounds using chemical synthesis and test the compounds using patient-derived stem cells for improvements in cellular assays. These new derivative compounds should create a more extensive SAR and will be the next generation compounds to develop more lead-like compounds for AD therapeutics.

项目摘要/摘要 该项目的长期目标是开发治疗阿尔茨海默氏症的疗法，以抑制一种独特的 细胞死亡称为坏死性下垂。已知这一细胞死亡途径在 神经退行性疾病，强调了找到针对相关蛋白质的治疗方法的必要性。几个 已知的激酶是重要的调节因子，包括：RIPK1、RIPK3和MLKL。然而，与大多数不同的是 针对与坏死性下垂有关的RIPK蛋白的激酶部分的治疗方法，我们的目标是 RIPK3和MLKL之间的蛋白质-蛋白质界面。这种定向方法应该会揭示出更具体的 自RIPK激酶以来的化合物参与了多个途径。通过以仅共享的接口为目标 在涉及坏死性下垂的两个蛋白质之间(蛋白质-蛋白质界面)，我们确保有更少的侧边 与以激酶功能为靶点相比，其影响不仅仅涉及坏死性下垂。以前是这样的 结果表明，RIPK3和远离激酶界面的MLKL之间的相互作用对于 促进RIPK3和MLKL绑定。我们在Silico中瞄准了参与RIPK3交互的这个口袋 使用小分子文库的MLKL。对前13种化合物进行了流式细胞分析，其中 细胞凋亡受到抑制，细胞仅通过坏死性下垂经历细胞死亡。使用这种分析方法，我们能够 将化合物列表缩小到两种特定抑制坏死性下垂的化合物，我们展示了这些化合物 以细胞中的RIPK3蛋白为靶点。对这两种Hit化合物的衍生物的后续研究显示 改进。我们建议通过化学合成进一步开发化合物，并对化合物进行测试 使用患者来源的干细胞来改进细胞分析。这些新的衍生化合物应该 创造更广泛的SAR，并将是下一代化合物开发更多的类似铅的化合物 用来治疗AD的。

项目成果

Generation of an iPSC line from a Pontocerebellar Hypoplasia 1B patient harboring a homozygous c.395 A > C mutation in EXOSC3 along with a family matched control.

• DOI: 10.1016/j.scr.2022.102944
• 发表时间: 2022-12
• 期刊: STEM CELL RESEARCH
• 影响因子: 1.2
• 作者: Stansfield, Ben N.;Rangasamy, Sampath;Ramsey, Keri;Khanna, May;Churko, Jared M.
• 通讯作者: Churko, Jared M.