Engineering selective metalloproteinases inhibitors for developing new therapeutics for neurodegenerative diseases

工程选择性金属蛋白酶抑制剂用于开发神经退行性疾病新疗法

• 批准号: 10194527
• 负责人: Maryam Raeeszadeh Sarmazdeh
• 金额: $ 24.11万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-05 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10194527
• 关键词: Affinity; Alzheimer' s Disease; Binding; Biological Markers; Complex; Directed Molecular Evolution; Disease; Disintegrins; Engineering; Enzyme Inhibitor Drugs; Enzymes; Foundations; Gelatinase B; Goals; Growth Factor; Huntington Disease; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Metalloproteases; Modeling; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurosciences; Parkinson Disease; Pathogenicity; Pathologic Processes; Physiological Processes; Play; Protein Engineering; Proteins; Role; Scaffolding Protein; Surface; TIMP3 gene; Therapeutic; Tissue Engineering; Variant; X-Ray Crystallography; Yeasts; base; drug candidate; improved; in vivo Model; inhibitor/antagonist; novel; novel therapeutic intervention; novel therapeutics; overexpression; pre-clinical; scaffold; therapeutic enzyme; therapeutic target

Targeting new pathogenic biomarkers of neurodegenerative diseases is critical as there are limited efficient therapeutics available to control such diseases. The metzincin superfamily, including matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinases (ADAMs), play multifaceted roles in physiological and pathological processes in the central nervous system and therefore are therapeutic targets to limit neurodegeneration in diseases such as Huntington disease, Parkinson’s disease (PD), and Alzheimer’s disease (AD). Given the significance of recognizing enzymes that play a central role in neurodegenerative disease as novel neurodegenerative therapeutics, enzyme inhibitors with high selectivity are desired. Overexpression of MMP-9 plays a significant role in several neurodegenerative disorders, while ADAM-10 helps block progression of AD. Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a natural inhibitor of MMP-9 with pico- and subnanomolar affinity. To overcome the challenges of wide multispecificity of TIMP-3 for different classes of MMPs and ADAMs and its interaction with growth factors, a protein engineering approach is desired to tailor a TIMP-3 scaffold to create an outstanding neurodegenerative drug candidate. The overarching goal of this proposal is to engineer protein-based scaffolds that high selectively target a specific metzincin without off-target effects can create novel therapeutics for neurodegenerative diseases. In Aim 1, we propose to engineer protein scaffolds based on TIMPs to improve binding selectivity toward MMP-9 using directed evolution and yeast surface display. In Aim 2, we will study the mechanism of interaction of these TIMP-3 variants in complex with MMP-9 and ADAM-10 using X-ray crystallography to understand the underlying mechanism of inhibition. In Aim 3, we will evaluate these engineered TIMP-3 scaffolds with improved selectivity for MMP-9 in neurodegenerative models. Our long-term goal is to develop selective protein-based therapeutics based on MMP inhibitors or similar scaffolds. These studies will lay the foundation for preclinical in vivo models and novel therapeutic strategies for neurodegenerative and other MMP-related diseases.

靶向神经退行性疾病的新致病生物标志物至关重要，因为可用于控制此类疾病的有效治疗方法有限，包括基质金属蛋白酶 (MMP) 和解整合素和金属蛋白酶 (ADAM)，在神经退行性疾病的生理和病理过程中发挥多方面的作用。因此，它是限制亨廷顿病、帕金森病 (PD) 和阿尔茨海默病 (AD) 等疾病的神经退行性变的治疗靶点。认识到在神经退行性疾病中发挥核心作用的酶作为新型神经退行性疗法的重要性，需要具有高选择性的酶抑制剂，MMP-9 的过度表达在多种神经退行性疾病中发挥重要作用，而 ADAM-10 有助于阻止 AD 的进展。金属蛋白酶组织抑制剂-3 (TIMP-3) 是 MMP-9 的天然抑制剂，具有皮摩尔和亚纳摩尔亲和力，可克服 TIMP-3 对不同细胞具有广泛多特异性的挑战。为了了解 MMP 和 ADAM 的类别及其与生长因子的相互作用，需要一种蛋白质工程方法来定制 TIMP-3 支架，以创建出色的神经退行性疾病候选药物。 该提案的总体目标是设计基于蛋白质的支架，高选择性地靶向特定的 metzincin，而不会产生脱靶效应，从而为神经退行性疾病创造新的疗法。在目标 1 中，我们建议设计基于 TIMP 的蛋白质支架，以提高结合选择性。使用定向进化和酵母表面展示实现 MMP-9 在目标 2 中，我们将使用 MMP-9 和 ADAM-10 研究这些 TIMP-3 变体的相互作用机制。通过 X 射线晶体学来了解抑制的潜在机制 在目标 3 中，我们将评估这些工程化 TIMP-3 支架，在神经退行性模型中对 MMP-9 具有更高的选择性。这些研究将为神经退行性疾病和其他 MMP 相关疾病的临床前体内模型和新治疗策略奠定基础。