Turbo Eye Drops to Treat Ocular Toxicity and Blindness from Sulfur Mustard

涡轮滴眼液治疗硫芥引起的眼部毒性和失明

• 批准号: 10015700
• 负责人: Rajiv Ravindra Mohan
• 金额: $ 70.3万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10015700
• 关键词: Acute; Adverse effects; Affect; Alkylating Agents; Anatomy; Angiogenic Factor; Antidotes; Basement membrane; Biological Assay; Blindness; Cities; Clinical; Clinical Pathology; Collagen Fibril; Consult; Cornea; Corneal Injury; Corneal Ulcer; Data; Development; Diagnostic; Diagnostic Imaging; Dose; Drops; Electron Microscopy; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelium; Euthanasia; Extracellular Matrix Degradation; Eye; Eye Injuries; Eyedrops; FDA approved; Fibrosis; Fluorescein; Formulation; Frequencies; Functional disorder; Funding; Generic Drugs; Goals; Grant; Health; Histologic; Human; Immunofluorescence Immunologic; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Intellectual Property; Iran; Iraq; Joints; Kansas; Keratopathy; Lead; Letters; Literature; Magnetic Resonance Imaging; Measures; Mediating; Methods; Microscope; Modeling; Molecular; Monitor; Mustard Gas; Ocular Pathology; Ophthalmic Solutions; Ophthalmic examination and evaluation; Ophthalmologist; Organ Culture Techniques; Oryctolagus cuniculus; Pain; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Pilot Projects; Process; Production; Property Rights; Publications; Publishing; Recurrence; Regimen; Research; Safety; Schedule; Scientist; Secure; Syria; Techniques; Technology; Temperature; Terrorism; Testing; Therapeutic Effect; Time; Tissues; Toxic effect; Transmission Electron Microscopy; Treatment Efficacy; Treatment Protocols; Vesicants; Veterinarians; Vision; War; Western Blotting; World War I; base; bench to bedside; corneal epithelium; drug development; effective therapy; high risk; in vivo; in vivo evaluation; limbal; medical countermeasure; meetings; multimodality; neovascularization; novel; side effect; stem cells; tomography; tonometry; tool; vapor

ABSTRACT Sulfur mustard gas (SM), a vesicating and warfare agent, has been used in many wars since World War I; most recently in Syria. SM rapidly penetrates the eye on contact and causes blindness by injuring corneal tissue- organization and function. Clinically, patients show a pathology termed as Mustard Gas Keratopathy that involves severe ocular inflammation, recurrent epithelial-erosions, epithelial-stromal separation limbal stem cell deficiency, corneal ulceration, haze and neovascularization. MGK pathophysiology is biphasic including acute and delayed-onset, and involves multiple mechanisms. We developed a novel, multimodal, non-steroidal topical ophthalmic drops, Turbo Eye Drop (TED), containing 4 FDA-approved generic drugs with differing mode of action, and stable at ambient temperature. Our pilot studies found that topical TED efficaciously treats acute and delayed-onset MGK in rabbits in vivo and human cornea ex vivo without significant side effects. Our central hypothesis is that topical TED treats acute and delayed-onset MGK in vivo by curbing SM-induced early inflammatory responses, extracellular matrix degradation, and production of excessive pro-fibrotic and pro- angiogenic factors without significant side effects. This project tests two novel hypotheses to establish an efficacious and safe topical therapy for acute and delayed-onset MGK in vivo, using four specific aims: Aim-1 defines TED treatment for acute MGK in vivo by testing the hypothesis that increasing frequency and duration of TED application will potently treat acute MGK and blindness without significant side effects. Aim-2 establishes TED treatment for delayed-onset MGK in vivo by testing the hypothesis that low TED topical dosing for longer duration will effectively cure delayed-onset MGK without issues in rabbits. Aim-3 uncovers mechanisms used by TED in mitigating acute and delayed-onset MGK in vivo and in vitro. Aim-4 secures intellectual property rights, develops regulatory strategies, and advances TED topical ophthalmic drops as an antidote for SM-induced ocular injury towards human application. This will be accomplished using an established SM-vapor rabbit in vivo and human cornea organ culture ex vivo models, GMP-grade TED eye drops, and monitoring eyes in live rabbits in a time-dependent manner with clinical eye exams and diagnostic imaging. The characterization of mechanisms used by TED in mitigating MGK will be studied using corneal tissues collected after euthanasia by measuring integrity of corneal epithelial basement membrane, epithelial-stromal organization, and collagen fibril arrangement using qPCR, ELISAs, immunofluorescence, H&E, and transmission electron microscopy techniques utilizing our published methods. Successful completion of the project will lead to the development of an effective and safe therapy for acute and delayed MGK and medical countermeasure to minimize ocular obliteration caused by the accidental or intentional use of SM in humans, and therefore will have very high impact in field and public safety.

抽象的 硫芥子气 (SM) 是一种起泡剂和战剂，自第一次世界大战以来已在许多战争中使用；最多 最近在叙利亚。 SM 在接触时迅速穿透眼睛并通过损伤角膜组织导致失明 - 组织和职能。临床上，患者表现出一种称为芥子气角膜病的病理学，涉及 严重眼部炎症、复发性上皮糜烂、上皮基质分离角膜缘干细胞 缺乏，角膜溃疡，混浊和新生血管。 MGK 病理生理学是双相的，包括急性 且迟发，并涉及多种机制。我们开发了一种新型、多模式、非类固醇外用药物 滴眼液，Turbo Eye Drop (TED)，含有 4 种 FDA 批准的仿制药，具有不同的作用模式 作用，并且在环境温度下稳定。我们的初步研究发现，局部 TED 可以有效治疗急性和 迟发性 MGK 在兔体内和离体人角膜中均无明显副作用。我们的中央 假设局部 TED 通过抑制 SM 诱导的早期症状来治疗体内急性和迟发性 MGK 炎症反应、细胞外基质降解以及过度促纤维化和促纤维化的产生 血管生成因子，无明显副作用。该项目测试了两个新颖的假设，以建立一个 针对急性和迟发性 MGK 的体内有效且安全的局部治疗，采用四个具体目标：Aim-1 通过测试增加频率和持续时间的假设，定义了体内急性 MGK 的 TED 治疗 TED 的应用将有效治疗急性 MGK 和失明，且没有明显的副作用。 Aim-2 建立 通过测试低 TED 局部剂量持续更长时间的假设，对体内迟发性 MGK 进行 TED 治疗 持续时间将有效治愈迟发性 MGK，不会对兔子造成任何问题。 Aim-3 揭示了所使用的机制 TED 在体内和体外减轻急性和迟发性 MGK 的作用。 Aim-4 确保知识产权， 制定监管策略，并推进 TED 局部滴眼液作为 SM 诱发的解毒剂 对人体应用造成眼部损伤。这将使用已建立的 SM-vapor 兔子体内来完成 和人类角膜器官离体培养模型、GMP 级 TED 滴眼液以及活体兔子眼睛监测 以与时间相关的方式进行临床眼科检查和诊断成像。的表征 TED 用于减轻 MGK 的机制将使用安乐死后收集的角膜组织进行研究 测量角膜上皮基底膜、上皮基质组织和胶原纤维的完整性 使用 qPCR、ELISA、免疫荧光、H&E 和透射电子显微镜进行排列 利用我们发布的方法的技术。该项目的成功完成将带动 针对急性和迟发性 MGK 的有效且安全的治疗方法以及尽量减少眼部损伤的医疗对策 由于人类无意或有意使用 SM 造成的抹除，因此会产生非常高的影响 在现场和公共安全。