DEVELOPMENT OF A NOVEL ANTIVIRAL TO TREAT AND PREVENT ACYCLOVIR RESISTANCE IN HUMAN OCULAR HERPES KERATITIS

开发一种新型抗病毒药物来治疗和预防人眼疱疹性角膜炎的阿昔洛韦耐药性

基本信息

批准号：
9255235
负责人：
ROBERT Paul RICCIARDI
金额：
$ 30万
依托单位：
FOX CHASE CHEMICAL DIVERSITY CENTER, INC
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9255235
关键词：
Acute Acyclovir Animal Model Antiviral Agents Area Benzamides Binding Biological Assay Blindness Calorimetry Cell Proliferation Cells Cornea Crystallization DNA DNA Synthesis Inhibition DNA biosynthesis DNA-Directed DNA Polymerase Development Docking Drug Delivery Systems Drug Transport Drug resistance Epithelial Cells Escape Mutant Experimental Designs Exposure to Eye Infections Failure Family Felidae Future General Population Goals Gold HSV-Tk Gene Herpes Simplex Infections Herpesviridae Herpesviridae Infections Herpesvirus 1 Herpetic Keratitis Histology Human In Vitro Individual Infection Keratitis Lead Link Modeling Mutation Oral Patients Pennsylvania Permeability Pharmaceutical Chemistry Pharmaceutical Preparations Phase Polymerase Polymerase Gene Positioning Attribute Process Property Proteins Recurrence Resistance Resort Scanning Schools Shapes Solubility Stratified Epithelium Structure Sulfones Surface Plasmon Resonance Testing Therapeutic Index Time Tissue Model Tissues Titrations Toxic effect Universities Validation Vero Cells Viral Virus Vision analog base corneal epithelium corneal scar design experience flexibility genital herpes improved mutant novel prevent prophylactic protein transport residence resistance mechanism scaffold standard care standard of care thymidine kinase 1 viral DNA

项目摘要

ABSTRACT Infection of the eye by Herpes Simplex Virus-1 (HSV-1) can result in Herpes Keratitis (HK), which is the leading cause of corneal blindness worldwide. In the U.S., nearly 500,000 individuals experience ocular herpes infections that are often recurrent and culminate in progressive corneal scarring and loss of vision. The gold standard of care for HK is treatment with Acyclovir (ACV) that targets HSV-1 thymidine kinase (TK). Although ACV is extremely effective in both oral and genital herpes with negligible drug failure, the emergence of viral resistant mutants in 7-14% of ocular HK patients is compelling. Significantly, the mechanism of drug resistance is directly related to mutational alterations in the TK gene of HSV-1 isolated from HK patients unresponsive to ACV. This high level of resistance can be prevented if ACV is combined with a second antiviral directed against a different target to block mutants that escape either drug. One new class of antiviral targets is the processivity factors (PFs) that are essential to keep their cognate viral polymerases (Pols) tethered to the template for continuous viral DNA synthesis. Our objective is to develop early lead antiviral compounds that are directed against the HSV-1 processivity factor (PF). We have now identified a potent early Lead Z9445, which blocks infection of HSV-1 with an EC50 of 280 nM. We have also identified structurally diverse backup leads. With the future requirement of animal model validation, we are in a unique position to treat multiple feline patients (Phase 2) from the UPenn Vet School who are infected with the homologous Feline Herpes Virus-1 (FHV-1). These feline patients provide a close and natural model of human herpes keratitis that can be followed for extended periods for recurrent infections. We have now cloned the FHV-1 PF/Pol genes which share considerable homologies to those of HSV-1 and have validated that the majority of our compounds that block HSV-1, also block FHV-1 infection with similar potencies. Even though the feline model is highly attractive, our experimental design is focused, such that analogs that are singly superior for HSV-1 (but not FHV-1) will be developed further (Phase 2) using traditional models. The Aims of this proposal are tightly linked, employing medicinal chemistry as an iterative process to improve the therapeutic index of Z9445 by generating new analogs that have increased potency with no detectable toxicities. Rational design based on docking of Z9445 to the known crystal structure of the PF target protein will be one means on producing new Leads. Backup Leads will also be optimized should properties such as solubility or stability need to be enhanced (Phase 2). Analogs will be tested for blocking processive DNA synthesis in vitro, physical binding to the PF target protein, toxicity and cell proliferation, antiviral activities in primary feline corneal epithelial cells and in the human 3D corneal tissue. The human 3D corneal tissue will be tested further for permeability and histology. .

抽象的单纯疱疹病毒 1 (HSV-1) 感染眼睛可导致疱疹性角膜炎 (HK)，这是全世界角膜失明的主要原因。在美国，近 50 万人经历过眼部疱疹感染经常复发并最终导致进行性角膜疤痕和丧失的视觉。香港护理的黄金标准是针对 HSV-1 的阿昔洛韦 (ACV) 治疗胸苷激酶（TK）。尽管苹果醋对于口腔和生殖器疱疹都非常有效药物失败可以忽略不计，7-14% 的眼 HK 患者中出现病毒耐药突变体引人注目的。值得注意的是，耐药机制与突变改变直接相关存在于对 ACV 无反应的香港患者中分离出的 HSV-1 的 TK 基因中。这种高水平的阻力如果将 ACV 与另一种针对不同靶点的抗病毒药物联合使用来阻断，则可以预防逃避这两种药物的突变体。一类新的抗病毒靶标是持续性因子（PF），对于将同源病毒聚合酶 (Pols) 固定在模板上以实现连续病毒传播至关重要 DNA合成。我们的目标是开发针对病毒的早期先导抗病毒化合物 HSV-1 持续因子 (PF)。我们现在已经鉴定出一种有效的早期先导物 Z9445，它可以阻断 HSV-1 感染，EC50 为 280 nM。我们还确定了结构多样化的备用线索。随着未来动物模型验证的要求，我们处于独特的地位来治疗多种疾病来自宾夕法尼亚大学兽医学院的猫科动物患者（第二阶段）感染了同源猫科动物疱疹病毒-1 (FHV-1)。这些猫科动物患者提供了人类疱疹的接近和自然的模型角膜炎，可长期跟踪复发性感染。我们现在已经克隆了 FHV-1 PF/Pol 基因与 HSV-1 具有相当大的同源性，并已证实我们的大多数阻断 HSV-1 的化合物也能以类似的效力阻断 FHV-1 感染。尽管猫科动物模型非常有吸引力，但我们的实验设计是有针对性的，以便类似物单独优于 HSV-1（但不是 FHV-1）的药物将使用传统方法进一步开发（第 2 阶段）模型。该提案的目标是紧密相连的，采用药物化学作为迭代通过生成新的类似物来提高 Z9445 治疗指数的过程没有可检测到的毒性的效力。基于Z9445与已知晶振对接的合理设计 PF目标蛋白的结构将是产生新先导化合物的一种手段。备用线索也将如果需要增强溶解度或稳定性等特性（第 2 阶段），则需要对其进行优化。类似物将进行体外阻断 DNA 持续合成、与 PF 靶蛋白物理结合的测试，原代猫角膜上皮细胞和人类的毒性和细胞增殖、抗病毒活性 3D 角膜组织。人类 3D 角膜组织将进一步进行渗透性和组织学测试。。