Therapeutic Corneal Cross-linking Using Aliphatic Beta-Nitroalcohols

使用脂肪族 β-硝基醇进行治疗性角膜交联

• 批准号: 8616069
• 负责人: DAVID C PAIK
• 金额: $ 39.45万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8616069
• 关键词: Alcohols; Apoptosis; Benzalkonium Chloride; Biochemical; Biological Assay; Blindness; Body Temperature; Cell Proliferation; Cells; Chemicals; Chemistry; Collagen; Complication; Confocal Microscopy; Contact Lenses; Cornea; Corneal Diseases; Corneal Stroma; Corneal Topography; Debridement; Devices; Diffusion; Disease; Dose; Drug Delivery Systems; Drug Formulations; Endothelial Cells; Epidemiology; Epithelial; Epithelial Cells; Evaluation; Excision; Eye; Eyedrops; Failure; Funding; Future; German population; Glycols; Grant; Histology; Human; Hydrogels; In Situ; In Situ Nick-End Labeling; In Vitro; Infection; Keratoconus; Laser In Situ Keratomileusis; Lasers; Lead; Life; Mechanics; Medicine; Merocel; Methods; Mono-S; Necrosis; Nitro Compounds; Operative Surgical Procedures; Oryctolagus cuniculus; Pain; Patients; Peripheral; Permeability; Phase I Clinical Trials; Physiological; Porifera; Preparation; Procedures; Propidium Diiodide; Reaction; Regimen; Research; Riboflavin; Risk; Safety; Simulate; Techniques; Technology; Temperature; Testing; Tetracaine; Therapeutic; Time; Tissues; Topical application; Toxic effect; Translations; Trypan Blue; Ultrasonography; Ultraviolet Rays; Work; annexin A5; catalyst; cell growth; corneal epithelium; crosslink; cytotoxicity; design; in vivo; instrumentation; keratomileusis; light transmission; member; new technology; nitration; public health relevance; reaction rate; research study; response; success; tonometry; ultraviolet irradiation

DESCRIPTION (provided by applicant): A major breakthrough in the treatment of keratoconus and post-LASIK keratectasias has been realized. Recent work by the German group of Wollensak, Spoerl, and Seiler has shown that cross-linking corneal collagen through application of riboflavin and ultraviolet light (UVAR) can limit progressive vision loss in keratoconus patients. Despite these successes, the UVAR therapy poses attendant risks, particularly related to ultraviolet irradiation, is prohibited in thin corneas, and requires painful epithelial removal. A better approach may be possible. The thrust of this proposal is to develop an alternative method of corneal collagen cross- linking using novel technology. In particular, we have identified a class of compounds (i.e. aliphatic ?-nitro alcohols = BNAs) that appear to be safe and effective as tissue cross-linking agents under conditions of physiologic pH and temperature. Potential advantages over current UVAR therapy include the omission of ultraviolet light exposure, the ability to treat thin corneas, no epithelial debridement resulting in less patient discomfort and a lower chance of infection, a dose-response effect, and the ability to treat the peripheral cornea. This project is designed to lay groundwork for rapid translation of this technology into a treatment. In aim 1, the chemical mechanisms involved in BNA cross-linking reactions will be determined using analytical instrumentation. Mechanistic determinations will lead to the identification of effective catalysts which will be confirmed using biochemical shrinkage temperature analysis and mechanical failure testing. Aim 1 is designed to identify one or more "candidate eye drop preparations" that will then be used for in vivo rabbit experiments. In aim 2, primary cultures of corneal epithelial cells, keratocytes, and endothelial cells will be used to determine the toxic thresholds for BNAs (including higher order BNAs = HONAs) and catalysts. The results from this aim will establish the toxicity level of nitro compounds to corneal cells. In aim 3, BNA transcorneal permeability using a Franz diffusion cell and ex vivo rabbit corneas will be determined. In addition, ways of optimizing permeability (enhancing agents such as proparacaine and benzalkonium chloride) and delivery (devices such as merocel sponges, hydrogel contact lenses, and collagen shields) will be studied. In aim 4, topical cross-linking of live rabbits will be undertaken. The delivery method and agents used will be determined by the information gained from aims 1, 2, and 3. Real time in vivo efficacy and safety evaluations (using in vivo confocal microscopy, ultrasound pachymetry, corneal topography, and applanation tonometry) will be performed, followed by post-mortem mechanical failure testing, shrinkage temperature analysis, histology, and TEM. The results of this aim will establish the in vivo efficacy and safety of this technology and will dictate the feasibility of a human phase I trial.

描述（由申请人提供）：已经实现了圆锥角膜和lasik肾上腺切开胸骨的重大突破。德国沃伦萨克（Wollensak），斯波尔（Spoerl）和塞勒（Seiler）的最新工作表明，通过应用核黄素和紫外线（UVAR）通过涂抹角膜胶原蛋白（UVAR）可以限制角膜核患者的渐进视力丧失。尽管取得了这些成功，但UVAR疗法仍会在薄的角膜中构成随之而来的风险，尤其是与紫外线照射有关的风险，并且需要疼痛的上皮去除。更好的方法可能是可能的。该提案的目的是开发一种使用新技术的角膜胶原交叉联系的替代方法。特别是，在生理pH和温度条件下，我们已经确定了一类化合物（即脂肪族？-Nitro醇= BNA），它们似乎是安全有效的，它们是组织交联剂。与当前UVAR疗法相比，潜在的优势包括省略紫外线光暴露，治疗薄膜的能力，没有上皮清创术，导致患者不适和感染的机会较低，剂量反应效应以及治疗外围角膜的能力。该项目旨在为将该技术的快速转换为一种治疗奠定基础。 在AIM 1中，将使用分析仪器确定参与BNA交联反应的化学机制。机械测定将导致鉴定有效的催化剂，这些催化剂将通过生化收缩温度分析和机械衰竭测试来确认。 AIM 1旨在识别一种或多种“候选眼滴剂制剂”，然后将其用于体内兔实验。在AIM 2中，将使用角膜上皮细胞，角膜细胞和内皮细胞的原发性培养物来确定BNA（包括高阶BNA = Honas）和催化剂的毒性阈值。该目标的结果将确定硝基化合物对角膜细胞的毒性水平。在AIM 3中，将确定使用Franz扩散细胞和离体兔角膜的BNA跨核通透性。此外，将研究优化渗透性（增强丙帕菌和苯二烷基氯化物）和递送（诸如Merocel海绵，水凝胶隐形眼镜和胶原蛋白盾）的方式。在AIM 4中，将进行活兔子的局部交联。 The delivery method and agents used will be determined by the information gained from aims 1, 2, and 3. Real time in vivo efficacy and safety evaluations (using in vivo confocal microscopy, ultrasound pachymetry, corneal topography, and applanation tonometry) will be performed, followed by post-mortem mechanical failure testing, shrinkage temperature analysis, histology, and TEM.该目标的结果将确定该技术的体内功效和安全性，并决定人类I期试验的可行性。