Chemical Optimization of c-Cbl Antagonists for Corneal Wound Healing

用于角膜伤口愈合的 c-Cbl 拮抗剂的化学优化

基本信息

批准号：
10557187
负责人：
BRIAN P. CERESA
金额：
$ 38.5万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557187
关键词：
Acceleration Address Advanced Development Affinity Anatomy Animal Model Antineoplastic Agents Bacteria Binding Biochemical Biological Assay Biology Biomedical Engineering Blindness Bypass Calorimetry Cell Proliferation Cell model Cetuximab Chemical Injury Chemical Structure Chemicals Complex Cornea Corneal Diseases Corneal Injury Data Development Diabetes Mellitus Disease Distress Drug Side Effects Drug usage EGF gene Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Epithelial Cells Epithelium Event Eye Eye Infections FDA approved Generations Goals Growth Factor Receptors Health care facility Homeostasis Human In Vitro Incidence Infection Investigation Laboratories Lead Ligands Light Lysosomes Maintenance Measures Mediating Membrane Mission Modeling Modification Molecular Operative Surgical Procedures Oryctolagus cuniculus Pain Patients Prevalence Primary Care Procedures Proliferating Proteins Public Health Reagent Receptor Signaling Recombinants Recurrence Research Retina Rodent Signal Transduction Structure Testing Therapeutic Therapeutic Agents Tissue Model Tissues Topical application Trauma Virus Vision Work antagonist cancer type cell growth cell motility cell replacement therapy corneal epithelial wound healing corneal epithelium corneal surgery cytokine cytotoxicity desensitization drug development drug-like compound experience heat injury improved in silico in vivo inhibitor innovation interest knock-down limbal nanomolar novel novel therapeutics particle pharmacologic preservation prevent receptor restoration side effect stem cell replacement tissue culture trafficking ubiquitin-protein ligase wound healing

项目摘要

Project Summary An intact and fully differentiated corneal epithelium is critical for proper vision and to keep foreign objects (bacteria, viruses, small particles) out of the eye. However, damage to the corneal epithelium is one of the most common ocular problems presented in primary care facilities and arises from a variety of factors, including trauma, disease, and a side-effect of drugs. Despite the prevalence, discomfort, and potential for blindness associated with perturbation of the corneal epithelium, there are no FDA-approved agents that promote the restoration and homeostasis of this tissue. The long-term goal of our research is to develop new compounds that promote corneal epithelial wound healing and homeostasis. The overall objective of this application is to chemically optimize a recently identified c-Cbl antagonist to increase its affinity and potency, while decreasing cytotoxicity. To accomplish this, we will test the central hypothesis that compounds that disrupt EGFR:c-Cbl interactions will prevent EGFR ubiquitylation, divert the activated EGFR from the lysosome, and prolong EGFR signaling. Based on preliminary studies, this prolongs EGFR activity and increases the restoration and maintenance of corneal epithelium via cell migration, proliferation, and differentiation. The rationale for these studies is that knockdown of c-Cbl and inhibits EGFR ubiquitylation, prolongs EGFR activity, and enhances corneal epithelial wound healing. Following an in silico screen of 25,000,000 compounds, we have identified several lead compounds that bind c-Cbl and inhibit ligand-mediated EGFR ubiquitylation. We seek to develop the top compound into a therapeutic agent with the following specific aims. In Aim 1, we will structurally optimize our lead compound to increase its affinity for c-Cbl and disrupt EGFR:c-Cbl binding. Multiple rounds of chemical modifications to our lead compound will be used to identify new derivatives that bind recombinant c-Cbl with high affinity and disrupt EGFR:c-Cbl interactions. In Aim 2, we will test the highest affinity compounds for disruption of c-Cbl-mediated ubiquitylation and trafficking of the EGFR. The top candidates will be tested for their ability to block ligand-mediated EGFR ubiquitylation and lysosomal degradation, and if they prolong EGFR signaling in corneal epithelial cells. In Aim 3, we will determine whether inhibitors of EGFR ubiquitylation promote corneal epithelial wound healing. The top compounds will be assayed for corneal epithelial wound healing using in vitro, ex vivo, and in vivo assays. Our proposed studies are innovative, in our opinion, because we will by-pass the limitation of EGFR occupancy and target a novel protein (c-Cbl) and molecular mechanism (receptor desensitization). These studies are significant because accelerating re-epithelialization and homeostasis of compromised corneas will decrease patient distress, minimize infections, and reduce the incidence of blindness. Corneal epithelial homeostasis is a major public health issue with limited treatments. Our goal is to develop our lead compounds for the topical treatment of compromised corneal epithelium to restore tissue homeostasis.

项目摘要完整且完全不同的角膜上皮对于适当的视力至关重要，并保持异物（细菌，病毒，小颗粒）从眼睛中脱出。但是，对角膜上皮的损害是初级保健设施中最常见的眼部问题，是由多种因素引起的包括创伤，疾病和药物的副作用。尽管普遍存在，不适和潜力与角膜上皮扰动相关的失明，没有FDA批准的药物促进该组织的恢复和稳态。我们研究的长期目标是开发新的促进角膜上皮伤口愈合和稳态的化合物。总体目标应用是为了化学优化最近确定的C-CBL拮抗剂以提高其亲和力和效力，同时降低细胞毒性。为此，我们将测试中心假设，使得破坏EGFR：C-CBL相互作用将防止EGFR泛素化，将激活的EGFR从溶酶体转移，并延长EGFR信号。基于初步研究，这延长了EGFR活性并增加了通过细胞迁移，增殖和分化来恢复和维持角膜上皮。这这些研究的基本原理是敲除C-CBL并抑制EGFR泛素化，延长EGFR活性，并增强角膜上皮伤口愈合。在25,000,000种化合物的硅屏幕中，我们已经确定了几种结合C-CBL并抑制配体介导的EGFR泛素化的铅化合物。我们寻求将顶部化合物开发为具有以下特定目的的治疗剂。在AIM 1中，我们将从结构上优化我们的铅化合物，以增加对C-CBL的亲和力并破坏EGFR：C-CBL结合。对我们的铅化合物进行多轮化学修饰将用于识别新的衍生物结合具有高亲和力的重组C-CBL，并破坏EGFR：C-CBL相互作用。在AIM 2中，我们将测试最高 C-CBL介导的泛素化和EGFR贩运的相关化合物。顶部候选人将测试其阻断配体介导的EGFR泛素化和溶酶体的能力降解，如果它们延长了角膜上皮细胞中的EGFR信号传导。在AIM 3中，我们将确定是否 EGFR泛素化的抑制剂可促进角膜上皮伤口愈合。顶级化合物将是测定使用体外，体内和体内测定的角膜上皮伤口愈合。我们提出的研究在我们看来，具有创新性，因为我们将绕过EGFR入住率的局限性并针对小说蛋白质（C-CBL）和分子机制（受体脱敏）。这些研究很重要，因为加速重新上皮化和受损角膜的稳态将减少患者的困扰，最大程度地减少感染，并减少失明的发生率。角膜上皮稳态是主要的公众健康问题有限治疗。我们的目标是开发我们的铅化合物，以局部处理损害了角膜上皮，以恢复组织稳态。