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

基本信息

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
项目类别：
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)

项目摘要

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/摘要增殖性糖尿病性视网膜病，与年龄相关的黄斑变性和预性视网膜病变均为眼睛的疾病会导致失明，并且由于视网膜或脉络膜血液异常发育引起船只。虽然靶向血管内皮生长因子信号传导的玻璃室内抗血管生成疗法通常对这些疾病有效，自发或获得的抵抗是一个重大问题，指出需要高质量的基于细胞的化学探针来审问血管生成途径和开发替代疗法。为了满足这种未满足的需求，我们建议开发基于高质量的细胞 profilin1（PFN1） - 肌动蛋白蛋白质相互作用的化学探针。 PFN1对于血管生成至关重要在响应血管生成信号的肌动蛋白细胞骨架的动态重塑中起着至关重要的作用。我们在许多情况下显示了抑制或抑制PFN1导致血管生成和最近证明，抑制PFN1降低了两个离体中新血管的形成和视网膜病的体内模型。我们已经确定了经过验证的命中化合物，该化合物抑制了生化和基于细胞的测定中的PFN1-肌动蛋白相互作用，并确认其在细胞中的靶标参与。到在维持类似药物的特性的同时，提高该抑制剂的效力，我们将采用迭代优化过程将由我们的结构和化学模型以及结构 - 在化合物的每次迭代期间，将围绕变异的关键点发展的活动关系选择，合成和生物测试。衍生物将在封闭式测定级联评估以确定它们对PFN1和细胞活性的结合亲和力。这种迭代过程旨在确定PFN1-的抑制剂肌动蛋白与生化和细胞测定中的亚微摩尔效力的相互作用。相遇的化合物在我们的第一轮测定中，定义明确的新颖性和效力标准将在第二系列中得到验证确认目标参与，选择性和其他功能公用事业的测定法（例如 VEGF代理和屏障功能调节剂）。这些研究成功完成将导致 PFN1-肌动蛋白的有效和特异性抑制剂用于研究PFN1在异常血管生成中的作用，并且可能最终导致治疗眼病的临床候选者。