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病原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有限。 Metzincin超家族，包括基质金属蛋白酶（MMP）以及崩解蛋白和金属蛋白酶（ADAMS），在中枢神经系统的物理和病理过程中起多方面的作用，因此是治疗靶点，因此是限制诸如Huntington病，帕克森病（Parkinson）疾病（Parkinsen and and and and and and pardine sealiser and parkins and ealimers and eal and eal and eal and eal and eal and eal and eal and）的疾病（p -pde）（pde）（pde）（pde）。鉴于识别酶在神经退行性疾病中起着核心作用作为新的神经退行性疗法的重要性，因此需要具有高选择性的酶抑制剂。 MMP-9的过表达在几种神经退行性疾病中起重要作用，而ADAM-10有助于阻止AD的进展。金属蛋白酶-3（TIMP-3）的组织抑制剂是具有Pico-和亚纳摩尔亲和力的MMP-9的天然抑制剂。为了克服TIMP-3对不同类别的MMP和ADAMS及其与生长因子的相互作用的挑战，需要采用蛋白质工程方法来量身定制TIMP-3脚手架，以创建出色的神经退行性药物候选药物。 该提案的总体目标是设计基于蛋白质的脚手架，这些支架高度选择性地靶向而没有脱靶效应的特定甲辛基可以为神经退行性疾病创造新的疗法。在AIM 1中，我们建议使用定向进化和酵母表面显示器基于TIMP来设计基于TIMP的蛋白质支架，以提高对MMP-9的结合选择性。在AIM 2中，我们将使用X射线晶体学研究这些TIMP-3变体与MMP-9和ADAM-10的相互作用的机理，以了解抑制的潜在机制。在AIM 3中，我们将在神经退行性模型中评估这些工程化的TIMP-3支架，对MMP-9的选择性提高。我们的长期目标是基于MMP抑制剂或类似脚手架开发基于选择性蛋白质的治疗。这些研究将为临床前体内模型和神经退行性和其他与MMP相关疾病的新型治疗策略奠定基础。