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）：圆锥角膜和 LASIK 术后角膜扩张的治疗已实现重大突破。德国 Wollensak、Spoerl 和 Seiler 小组最近的研究表明，通过应用核黄素和紫外线 (UVAR) 交联角膜胶原蛋白可以限制圆锥角膜患者进行性视力丧失。尽管取得了这些成功，UVAR 疗法也带来了伴随的风险，特别是与紫外线照射有关的风险，在薄角膜中被禁止，并且需要痛苦的上皮去除。也许有更好的方法。该提案的主旨是开发一种利用新技术进行角膜胶原交联的替代方法。特别是，我们已经确定了一类化合物（即脂肪族 α-硝基醇 = BNA），它们在生理 pH 和温度条件下作为组织交联剂似乎是安全有效的。与当前 UVAR 疗法相比，潜在优势包括省略紫外线照射、能够治疗薄角膜、无需上皮清创，从而减少患者不适和降低感染机会、剂量反应效应以及能够治疗周边角膜。该项目旨在为将该技术快速转化为治疗方法奠定基础。 在目标 1 中，将使用分析仪器确定 BNA 交联反应中涉及的化学机制。机械测定将导致有效催化剂的鉴定，这将通过生化收缩温度分析和机械故障测试进行确认。目标 1 旨在鉴定一种或多种“候选滴眼剂制剂”，然后将其用于兔子体内实验。在目标 2 中，角膜上皮细胞、角膜细胞和内皮细胞的原代培养物将用于确定 BNA（包括高阶 BNA = HONA）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。在目标 3 中，将使用 Franz 扩散池和离体兔角膜来确定 BNA 经角膜的渗透性。此外，还将研究优化渗透性（丙美卡因和苯扎氯铵等增强剂）和输送（merocel 海绵、水凝胶隐形眼镜和胶原蛋白防护罩等装置）的方法。在目标 4 中，将对活兔子进行局部交联。所使用的递送方法和药剂将根据从目标 1、2 和 3 获得的信息来确定。将进行实时体内功效和安全性评估（使用体内共焦显微镜、超声厚度测量、角膜地形图和压平眼压测量） ，然后进行事后机械故障测试、收缩温度分析、组织学和 TEM。这一目标的结果将确定该技术的体内功效和安全性，并将决定人体 I 期试验的可行性。