Directed evolution of tissue inhibitor of metalloproteinase 3 (TIMP-3) to develop novel Alzheimer’s disease (AD) therapeutics

金属蛋白酶组织抑制剂 3 (TIMP-3) 的定向进化可开发新型阿尔茨海默病 (AD) 疗法

• 批准号: 10303777
• 负责人: Maryam Raeeszadeh Sarmazdeh
• 金额: $ 30万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10303777
• 关键词: Affect; Affinity; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Alzheimer’s disease biomarker; American; Amyloid beta-Protein; Apoptosis; Binding; Biological Assay; Blood; Blood - brain barrier anatomy; Brain; Cell Culture Techniques; Clinical Research; Competitive Binding; Complex; Degenerative Disorder; Demyelinations; Dendrites; Directed Molecular Evolution; Disease; Disease Progression; Disintegrins; Engineering; Enzyme Inhibitor Drugs; Enzymes; Exposure to; Extracellular Matrix Degradation; Family; Future; Gelatinase B; Goals; Growth Factor; In Vitro; Inhibition of Matrix Metalloproteinases Pathway; Label; Libraries; Ligands; Light; MMP9 gene; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Metalloproteases; Mitochondria; Modeling; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Pathogenicity; Pathologic; Pathologic Processes; Pathway interactions; Peptide Hydrolases; Permeability; Physiological; Physiological Processes; Play; Protein Engineering; Proteins; Recombinants; Research; Research Personnel; Role; Scaffolding Protein; Signal Transduction; Signaling Molecule; Structure; Surface; TIMP3 gene; Testing; Therapeutic; Therapeutic Effect; Tissue Engineering; Tissue Inhibitor of Metalloproteinases; Tissues; Toxic effect; Up-Regulation; Variant; Work; X-Ray Crystallography; Yeasts; base; blood-brain barrier disruption; blood-brain barrier permeabilization; combinatorial; drug development; engineering design; enzyme mechanism; genetic variant; improved; improved outcome; inhibitor; insight; member; molecular targeted therapies; next generation; novel; novel therapeutics; rational design; scaffold; side effect; targeted treatment; tau aggregation; therapeutic target; tool; tumorigenesis

SUMMARY Limited efficient therapeutics are available to control Alzheimer's disease (AD), so there is a strong demand to target new pathogenic biomarkers of AD. The metzincin superfamily, including matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinases (ADAMs), play a multifaceted role in physiological and pathological processes in the central nervous system and therefore are therapeutic targets to limit neurodegeneration in AD. For instance, upregulation of MMP-9 promotes apoptosis, blood-brain barrier disorder, and demyelination and is involved in the amyloid-β processing pathway through proteolytic degradation of extracellular matrix components and tissue remodeling, and increasing tau accumulation which play key roles in AD. But on the other hand, upregulation of ADAM-10 increases soluble amyloid-β formation, which helps control AD. Thus, using highly selective inhibitors to tailor the proteolytic activity of metzincin- induced pathological changes in AD patients’ brains may be a promising therapeutic strategy. Protein inhibitors offer higher selectivity to target MMP-9 compared to smaller synthetic inhibitory molecules with broad-spectrum targets. Tissue inhibitors of metalloproteinases (TIMPs) are the major endogenous inhibitors of MMPs and ADAMs with varying degrees of binding affinity. Among all TIMPs, TIMP-3 has the lowest association with the cell signaling molecules triggering undesired off-target effect such as the ones responsible in tumorigenesis, suggesting that it has a low off-target therapeutic effect in MMP-dependent diseases. This makes TIMP-3 an ideal protein scaffold for engineering improved inhibitors for MMP-9. However, TIMP-3 also binds to other MMPs, ADAMs, and growth factors. Here, we propose to use a combination of rational and random combinatorial approaches to develop TIMP-3-based therapeutics to control neurodegenerative disease progression by targeting MMP-9 with high selectivity. These engineered TIMP-based probes will avoid binding to ADAM-10, which is neuroprotective in AD, to decrease off-target effects. The overarching goal of this project is to engineer and design a new molecularly targeted therapy based on natural MMP inhibitors that target MMP-9 with high selectivity, understand their mechanism of inhibition using structural studies and cell culture models to improve outcomes in neurodegenerative diseases with inadequate treatment options.

概括 可用于控制阿尔茨海默氏病（AD）的有效疗法有限，因此迫切需要 靶向 AD 的新致病生物标志物 metzincin 超家族，包括基质金属蛋白酶。 (MMP) 以及解整合素和金属蛋白酶 (ADAM)，在生理和功能方面发挥多方面的作用。 中枢神经系统的病理过程，因此是限制的治疗目标 例如，MMP-9 的上调可促进细胞凋亡、血脑屏障。 紊乱和脱髓鞘，并通过蛋白水解参与淀粉样蛋白-β 加工途径 细胞外基质成分的降解和组织重塑，以及增加 tau 积累，从而 但另一方面，ADAM-10 的上调会增加可溶性淀粉样蛋白-β 的形成， 这有助于控制 AD。因此，使用高度选择性的抑制剂来调整 metzincin 的蛋白水解活性。 诱导 AD 患者大脑发生病理变化可能是一种有前途的治疗策略。 与更小的广谱合成抑制分子相比，对目标 MMP-9 具有更高的选择性 金属蛋白酶组织抑制剂 (TIMP) 是 MMP 和 MMP 的主要内源性抑制剂。 在所有 TIMP 中，具有不同程度结合亲和力的 ADAM 与 TIMP-3 的关联性最低。 细胞信号分子触发不良的脱靶效应，例如导致肿瘤发生的分子， 这表明 TIMP-3 在 MMP 依赖性疾病中具有较低的脱靶治疗效果。 TIMP-3 是工程改良 MMP-9 抑制剂的理想蛋白质支架。 在这里，我们建议使用理性和随机的组合。 开发基于 TIMP-3 的疗法来控制神经退行性疾病的组合方法 这些基于 TIMP 的工程探针将通过高选择性靶向 MMP-9 来避免结合。 ADAM-10 对 AD 具有神经保护作用，以减少脱靶效应，这是该项目的总体目标。 是基于天然 MMP 抑制剂来设计和设计一种新的分子靶向疗法 MMP-9具有高选择性，通过结构研究和细胞培养了解其抑制机制 模型来改善治疗方案不足的神经退行性疾病的结果。