A new molecular therapy against ocular herpes

一种针对眼部疱疹的新分子疗法

• 批准号: 10363614
• 负责人: DEEPAK SHUKLA
• 金额: $ 46.22万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10363614
• 关键词: Acyclovir; Adverse effects; Affinity; Binding; Biochemical; Biological Availability; Blindness; Cell Cycle; Cells; Clinical; Complex; Contact Lenses; Cornea; Cyclodextrins; DNA; DNA biosynthesis; DNA delivery; Data; Development; Distress; Dose; Drug Combinations; Drug Kinetics; Drug resistance; Encephalitis; Engineering; Eye Infections; FRAP1 gene; Flow Cytometry; Formulation; Foscarnet; Funding; Ganciclovir; Glycoproteins; Goals; Graft Rejection; Herpesvirus 1; Herpetic Keratitis; Infection; Inflammation; Intravenous; Keratitis; Keratoplasty; Mass Spectrum Analysis; Measurement; Modeling; Modernization; Molecular; Morbidity - disease rate; Mus; Pathway interactions; Patients; Permeability; Pharmaceutical Preparations; Phosphorylation; Post-Translational Protein Processing; Prevention therapy; Prophylactic treatment; Protein Biosynthesis; Protein Synthesis Inhibitors; Proteins; Proteome; Proteomics; Publishing; Pyrrolidines; Refractory Disease; Reporting; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Simplexvirus; Solubility; Symptoms; Testing; Therapeutic; Thiophenes; Thymidine Kinase; Time; Tissues; Topical application; Toxic effect; Toxicology; Translations; Treatment Efficacy; Viral; Viral Proteins; Virus; Virus Shedding; Western Blotting; anti-viral efficacy; aptamer; aqueous; base; clinical care; corneal scar; design; disease prognosis; dosage; improved; in vivo; in vivo evaluation; inhibitor; mouse model; novel; nucleoside analog; ocular pain; pharmacokinetics and pharmacodynamics; prevent; prophylactic; receptor; recurrent infection; small molecule; sound; translational impact; viral DNA

Herpes simplex virus type-1 (HSV-1) with limited treatment options is a leading cause of infectious blindness and an important indication for corneal transplants in the US. The current treatment options include acyclovir and its derivatives, ganciclovir and foscarnet. All these drugs primarily act upon viral thymidine kinase to inhibit viral DNA replication and in essence have a similar mechanism of action. While these options are effective and show promise in reducing ocular HSV-1 infection including herpes stromal keratitis (HSK), emergence of drug resistance in the recent years has caused significant distress in the clinical care of ocular HSV-1 patients. As a result, there exists an unmet need for the development of new treatments against HSV-1 especially the ones that rely on novel modes of action. In our previous 3-year funding period we have characterized two candidates that target two different stages of HSV-1 infection; viral entry and viral protein synthesis. Against viral entry a highly effective anti-HSV-1 aptamer (DApt) was developed that targets HSV-1 glycoprotein D (gD). Similarly, we demonstrated for the first time that a small molecule PDK-1 inhibitor, BX795, blocks HSV-1 protein synthesis. Both candidates were tested in vivo for topical treatment of corneal HSV-1 infection and we reported significant improvements in disease prognosis. The major goals for the next funding period are to: (1) improve the in vivo efficacy of DApt, (2) understand the molecular basis of antiviral action by BX795, and (3) develop BX795 for systemic treatment of ocular herpes. In the first specific aim, we propose several ways to improve efficacy. We will engineer therapeutic contact lenses for sustained delivery of DApt, improve DApt formulation to include permeating agents such as cylodextrins to increase the depth to which the aptamer is delivered into the corneal tissue, and develop higher efficacy (synergistic or additive) drug combinations using DApt and BX795 or a nucleoside analog. In the second specific aim, we will decode the molecular mechanisms responsible for the antiviral activity of BX795 using both biased and unbiased approaches. Focus on the cap- dependent translation pathways responsible for viral protein synthesis will constitute the biased approach; cell- free and whole cell proteomic analyses using quantitative mass spectrometry and study of post translational modifications will constitute unbiased approaches. In the third specific aim, we will comprehensively study the pharmacokinetic and toxicological profiles of intravenously administered BX795. Using this data, we will finally understand the safe and effective dosage of BX795 required for the effective inhibition of primary and reactivated HSV-1 infection of the cornea using murine models of corneal infection. Taken together our studies will establish novel antiviral mechanisms and help design new prophylactic and therapeutic ways to control HSV-1 infection of the eye.

治疗选择有限的 1 型单纯疱疹病毒 (HSV-1) 是导致感染性失明的主要原因 也是美国角膜移植的重要适应症。目前的治疗方案包括阿昔洛韦 及其衍生物更昔洛韦和膦甲酸。所有这些药物主要作用于病毒胸苷激酶以抑制 病毒的复制在本质上与DNA有相似的作用机制。虽然这些选择是有效的并且 显示出减少眼部 HSV-1 感染（包括疱疹性基质角膜炎 (HSK)）的希望，药物的出现 近年来，耐药性给眼部 HSV-1 患者的临床护理带来了重大困扰。作为一个 因此，开发针对 HSV-1 的新疗法的需求尚未得到满足，尤其是 依赖于新颖的行动模式。在我们之前的 3 年资助期内，我们已经确定了两名候选人 针对 HSV-1 感染的两个不同阶段；病毒进入和病毒蛋白合成。防止病毒进入 开发了针对 HSV-1 糖蛋白 D (gD) 的高效抗 HSV-1 适体 (DApt)。相似地， 我们首次证明小分子 PDK-1 抑制剂 BX795 可以阻断 HSV-1 蛋白 合成。两种候选药物都经过了角膜 HSV-1 感染局部治疗的体内测试，我们报告了 疾病预后显着改善。下一个资助期的主要目标是：（1）改进 DApt 的体内功效，(2) 了解 BX795 抗病毒作用的分子基础，以及 (3) 开发 BX795 用于眼部疱疹的全身治疗。在第一个具体目标中，我们提出了几种改进方法 功效。我们将设计治疗性隐形眼镜以持续输送 DApt，改进 DApt 配方 包括渗透剂，例如环糊精，以增加适体递送到的深度 角膜组织，并使用 DApt 和开发更高功效（协同或相加）的药物组合 BX795 或核苷类似物。在第二个具体目标中，我们将解码分子机制 使用有偏见和无偏见的方法负责 BX795 的抗病毒活性。重点关注帽子—— 负责病毒蛋白合成的依赖翻译途径将构成有偏差的方法；细胞- 使用定量质谱法进行游离和全细胞蛋白质组分析以及翻译后研究 修改将构成公正的方法。在第三个具体目标中，我们将全面研究 静脉注射 BX795 的药代动力学和毒理学特征。使用这些数据，我们最终将 了解有效抑制原发性和 使用小鼠角膜感染模型重新激活角膜的 HSV-1 感染。综合我们的研究 将建立新的抗病毒机制并帮助设计新的预防和治疗方法来控制 眼睛 HSV-1 感染。