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.

具有有限治疗选择的单纯疱疹病毒类型1（HSV-1）是传染失明的主要原因 以及在美国角膜移植的重要迹象。当前的治疗方案包括阿西洛韦 及其衍生物Ganciclovir和Foscarnet。所有这些药物主要作用于病毒胸苷激酶以抑制 病毒DNA复制和本质上具有相似的作用机理。尽管这些选项有效，并且 在减少眼睛HSV-1感染（包括疱疹基质角膜炎（HSK））中，显示出有望 近年来，耐药性在眼部HSV-1患者的临床护理中造成了严重困扰。作为 结果，对针对HSV-1的新疗法的开发尤其是未满足的需求 依靠新颖的作用方式。在过去的三年资金期间，我们表征了两个候选人 该针对HSV-1感染的两个不同阶段；病毒进入和病毒蛋白质合成。反对病毒进入 开发了高效的抗HSV-1适体（DAPT），该抗HSV-1糖蛋白D（GD）。相似地， 我们首次证明了一个小的分子PDK-1抑制剂BX795阻止了HSV-1蛋白 合成。在体内测试了两种候选物，以局部治疗角膜HSV-1感染，我们报道 疾病预后的显着改善。下一个资金期间的主要目标是：（1）改进 DAPT的体内功效，（2）了解BX795的抗病毒作用的分子基础，并且（3）发展 BX795用于全身治疗眼疱疹。在第一个特定目标中，我们提出了几种改进的方法 功效。我们将设计治疗性隐形眼镜以持续交付DAPT，改善DAPT配方 包括包括渗透剂，例如cylodextrins，以增加适体的深度 角膜组织，并使用DAPT和 BX795或核苷类似物。在第二个特定目的中，我们将解码分子机制 使用偏见和无偏见的方法负责BX795的抗病毒活性。专注于帽子 负责病毒蛋白合成的依赖翻译途径将构成偏见的方法。细胞- 使用定量质谱法和翻译后的研究进行自由和全细胞蛋白质组学分析 修改将构成公正的方法。在第三个特定目标中，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 了解有效抑制初级和 使用角膜感染的鼠模型重新激活了角膜的HSV-1感染。总结我们的研究 将建立新型的抗病毒机制，并帮助设计新的预防性和治疗方法来控制 眼睛的HSV-1感染。