Discovery of Cell-based Chemical Probes Targeting Aberrant Angiogenesis in the Eye

发现针对眼部异常血管生成的基于细胞的化学探针

• 批准号: 10453044
• 负责人: Donna M Huryn
• 金额: $ 27.08万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10453044
• 关键词: 3-Dimensional; Actin-Binding Protein; Actins; Address; Affinity; Age related macular degeneration; Alternative Therapies; Binding; Biochemical; Bioinformatics; Biological Assay; Biological Testing; Blindness; Blood Vessels; Calorimetry; Cell Proliferation; Cells; Cellular Assay; Cellular biology; Chemicals; Choroid; Clinical; Computer Models; Critical Pathways; Cytoskeleton; Data; Development; Diabetic Retinopathy; Disease; Disease Progression; Dose; Drug Design; Endothelium; Evaluation; Exhibits; Eye; Eye diseases; Future; Goals; Heart; Image Analysis; In Vitro; Individual; Investigation; Lead; Ligation; Mediator of activation protein; Modeling; Modification; Morphogenesis; Ocular Pathology; Pathway interactions; Pharmaceutical Chemistry; Pharmaceutical Preparations; Play; Process; Property; Reproducibility; Resistance; Retina; Retinal Diseases; Retinopathy of Prematurity; Role; Route; Series; Signal Transduction; Structural Biochemistry; Structure; Structure-Activity Relationship; Synthesis Chemistry; Titrations; Toxic effect; Treatment Factor; Update; Validation; Variant; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; analog; angiogenesis; antagonist; base; cell motility; cheminformatics; clinical candidate; cytotoxicity; design; improved; in vivo; in vivo Model; inhibitor; mouse model; novel; novel therapeutics; ophthalmic drug; patient population; polymerization; prevent; process optimization; proliferative diabetic retinopathy; protein protein interaction; response; retinal angiogenesis; scaffold; screening; small molecule; structural biology; synergism; targeted treatment; thymosin beta(4)

Project Summary/Abstract Proliferative diabetic retinopathy, wet age-related macular degeneration, and retinopathy of prematurity are all diseases of the eye that can lead to blindness and are due to abnormal development of retinal or choroid blood vessels. Although intravitreal anti-angiogenic therapies targeting vascular endothelial growth factor signaling are generally effective for these diseases, spontaneous or acquired resistance is a significant problem and points to the need for high-quality cell-based chemical probes for interrogating angiogenic pathways and developing alternative therapies. To address this unmet need, we propose developing high-quality cell-based chemical probes for the profilin1 (Pfn1)-actin protein-protein interaction. Pfn1 is critical for angiogenesis as it plays a vital role in the dynamic remodeling of the actin cytoskeleton in response to angiogenic signals. We have shown in numerous contexts that inhibition or suppression of Pfn1 leads to reduced angiogenesis and have recently demonstrated that inhibiting Pfn1 reduces the formation of new blood vessels in both ex vivo and in vivo models of retinopathy. We have already identified a validated hit compound that inhibits the Pfn1-actin interaction in biochemical and cell-based assays and confirmed its target engagement in cells. To increase the potency of this inhibitor while maintaining drug-like properties, we will employ an iterative optimization process that will be guided by our structural and cheminformatic models and by the structure- activity relationship that will be developed around the key points of variation during each iteration of compound selection, synthesis, and biological testing. Derivatives will be evaluated in a gated assay cascade to determine their binding affinity for Pfn1 and activity in cells. This iterative process aims to identify an inhibitor of the Pfn1- actin interaction with sub-micromolar potency in both biochemical and cellular assays. Compounds that meet well-defined criteria for novelty and potency in our first round of assays will be validated in the second series of assays to confirm target engagement, selectivity, and other functional utilities (e.g., synergy with an anti- VEGF agent and barrier-function modulatory agent). Successful completion of these studies will result in a potent and specific inhibitor of Pfn1-actin for studying the role of Pfn1 in aberrant angiogenesis and may ultimately lead to a clinical candidate for the treatment of eye disease.

项目摘要/摘要 增殖性糖尿病视网膜病变、湿性老年性黄斑变性和早产儿视网膜病变都是 由于视网膜或脉络膜血液发育异常而导致失明的眼部疾病 船只。尽管针对血管内皮生长因子信号转导的玻璃体内抗血管生成治疗 通常对这些疾病有效，自发或获得性耐药是一个重大问题， 指出需要高质量的基于细胞的化学探针来询问血管生成途径和 开发替代疗法。为了解决这一未得到满足的需求，我们建议开发基于高质量细胞的 Profilin1(PFN1)-肌动蛋白-蛋白质相互作用的化学探针。PFN1对血管生成至关重要，因为它 在肌动蛋白细胞骨架对血管生成信号的动态重塑中起着至关重要的作用。我们 已经在许多背景下表明，抑制或抑制PFN1会导致血管生成减少和 最近证实，抑制PFN1在体外和体外都能减少新血管的形成 以及视网膜病变的活体模型。我们已经确定了一种有效的HIT化合物，它可以抑制 PFN1-肌动蛋白在生化和基于细胞的分析中的相互作用，并证实了其在细胞中的目标参与。至 在保持药物性质的同时增加这种抑制剂的效力，我们将使用迭代 优化过程将由我们的结构和化学信息学模型以及结构- 在化合物的每一次迭代中围绕变化关键点建立的活性关系 选择、合成和生物测试。衍生品将在门控分析级联中进行评估，以确定 它们与PFN1的结合亲和力和细胞内的活性。这一迭代过程旨在确定PFN1的抑制剂- 在生化和细胞分析中，肌动蛋白与亚微摩尔的相互作用。相遇的化合物 我们在第一轮检测中明确定义的新颖性和效力标准将在第二系列中得到验证。 确认目标结合、选择性和其他功能效用(例如，与抗病毒药物的协同作用)的分析 血管内皮生长因子制剂和屏障功能调节剂)。成功完成这些研究将导致 有效和特异的PFN1-肌动蛋白抑制剂用于研究PFN1在异常血管生成中的作用，并可能 最终形成治疗眼病的临床候选药物。