Optimizing a Stapled-Peptide That Specifically Targets HSV-1 to Treat Herpes Ocular Keratitis

优化专门针对 HSV-1 的钉合肽以治疗疱疹性眼角膜炎

基本信息

批准号：
10650858
负责人：
ROBERT Paul RICCIARDI
金额：
$ 99万
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：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10650858
关键词：
3-Dimensional Acyclovir Address Anatomy Antiviral Agents Area Binding Biological Assay Blindness Cells Chemicals Clinical Cornea Corneal Ulcer Crystallization Cyclic Peptides DNA DNA biosynthesis Development Disease Dissociation Dose Drug Kinetics Engineering Ensure Epithelial Cells Evaluation Exhibits Exposure to Eye Eye Infections Face Failure Formulation Gene Mutation Goals Herpes Labialis Herpesviridae Herpesviridae Infections Herpesvirus 1 Herpetic Keratitis Histopathology Human In Vitro Individual Infection Keratitis Lead Liposomes Measures Medical Modeling Mutagenicity Tests Oryctolagus cuniculus Patients Penetration Peptide Hydrolases Peptides Permeability Persons Pharmaceutical Preparations Physiological Polymerase Population Preclinical Drug Development Property Proteins Publishing Recurrence Research Resistance Risk Running Safety Series Solubility Solvents Surface Testing Therapeutic Time Tissues Toxic effect Vascularization Viral Viral Load result Virus Virus Replication Vision alpha helix analog anti-viral efficacy corneal epithelium corneal scar crosslink drug resistant virus efficacy study efficacy testing experience genital herpes genotoxicity in vivo mutant novel ocular surface peptide A pre-IND studies preclinical development prevent protein protein interaction residence risk mitigation safety testing scale up small molecule standard care stapled peptide three dimensional cell culture thymidine kinase 1 uptake 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 infectious 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 treatment is Acyclovir (ACV) that targets HSV-1 thymidine kinase (TK). However, emergence of ACV viral resistant mutants in 7-14% of ocular HK patients has created the urgent need to discover a second drug directed against a different HSV-1 target. A new class of antiviral targets are processivity factors (PFs) that are essential for tethering their cognate polymerases (Pols) to the template to enable continuous DNA synthesis. During HSV-1 replication, the DNA-anchored PF binds to the extreme C-terminus of Pol (C-Pol) to keep Pol from dissociating from the viral template. When co-crystallized with its PF, the C-terminus of the viral Pol (C-Pol) forms an α-helix, with one face making multiple bonds with several residues of PF while the other face is solvent exposed. Our goal is to develop a novel antiviral drug that specifically targets herpes PF for the purpose of treating ACV resistant HK. Stapled a-helical peptides have emerged for use in targeting protein-protein interactions that often display as long flat surfaces which are difficult for small molecules to bind efficiently. Peptide a-helices are ideal structural motifs for incorporating chemical staples to provide rigidity to make natural bonds with residues on the target protein. Moreover, staples can create a protease shield to prolong residence time. We have now engineered a stapled peptide of C-Pol (SPep7B) that blocks the mechanism of processive DNA synthesis in vitro and HSV-1 infection in human organotypic (3D) corneal epithelial cells; the unstapled peptide is non-inhibitory. SPep7B was shown to eliminate HSV-1 viral DNA in infected cells while failing to block a different virus. Importantly, SPep7B exhibits undetectable toxicity (CC50>100µM) by two different assays in human ocular epithelial cells. Our GOAL is to enter SPep7B into preclinical development as a Topical to treat HK. This will be accomplished by conducting a series of AIMS that include scaleup of SPep7B and evaluating two formulations for delivery of SPep7B into human 3D cultures and then ex-vivo excised rabbit cornea for permeability and toxicity. These studies are essential to confront the physiological and anatomical barriers to topical ocular delivery. In vivo studies will evaluate SPep7B for tolerability, genotoxicity, PK, and repeat dosing. An efficacy study will evaluate SPep7B for clinical cure and viral clearing. Backup analogs of SPep7B will be prepared and evaluated for risk mitigation. A model for new antiviral drugs: Closely aligned with this approach is the potential for using PFs and stapled peptides of other viruses for treating unmet medical needs caused by infections that lead to deleterious complications.

抽象的通过单纯疱疹病毒1（HSV-1）感染眼睛的眼睛会导致疱疹角膜炎（HK），这是全世界感染性角膜失明的主要原因。在美国，将近500,000个人体验经常复发和高潮的眼疱疹感染和视力丧失。黄金标准处理是针对HSV-1胸苷激酶的Acyclovir（ACV）（TK）。但是，在7-14％的眼HK患者中，ACV病毒抗性突变体的出现已创造迫切需要发现针对不同HSV-1靶标的第二种药物。一类新的抗病毒目标是加工性因子（PFS），对于将其同源聚合酶（POL）绑定到至关重要模板可以实现连续的DNA合成。在HSV-1复制期间，DNA锚定的PF结合到POL（C-POL）的极端C端，以防止POL与病毒模板分离。什么时候与其PF共结合，病毒POL（C-POL）的C末端形成α-螺旋，其中一个面部使一个脸多个键具有多个残留的PF，而另一个面则求解了暴露。我们的目标是发展一种新型的抗病毒药物，专门针对疱疹PF，目的是治疗抗ACV HK。已经出现了丁额A螺旋辣椒，用于靶向蛋白质 - 蛋白质相互作用，这些相互作用通常表现出长长的平坦表面，这对于小分子很难有效结合。肽A螺旋是合并化学主食以提供刚性以建立自然键的理想结构基序靶蛋白上残留物。此外，钉书钉可以创建一个蛋白酶屏蔽以延长住宅时间。现在，我们已经设计了C-POL（SPEP7B）的分阶段肽，该肽阻止了在人体有机（3D）角膜上皮细胞中的体外和HSV-1感染中的过程DNA合成；无抑制的肽是非抑制性的。显示SPEP7B消除了感染中HSV-1病毒DNA 细胞同时无法阻断不同的病毒。重要的是，SPEP7B表现出无法检测到的毒性（CC50> 100µm）通过人眼上皮细胞中的两个不同测定法。我们的目标是进入SPEP7B 作为治疗香港的局部发展，进入临床前发展。这将通过进行一系列目标，包括SPEP7B的规模以及评估将SPEP7B传递到中的两种形式人类3D培养物，然后出现前体激发兔角膜，以获得渗透性和毒性。这些研究是面对局部眼部递送的物理和解剖障碍至关重要。体内研究将评估SPEP7B的耐受性，遗传毒性，PK和重复给药。效率研究将评估用于临床治愈和病毒清除的SPEP7B。将准备和评估SPEP7B的备份类似物降低风险。一种新的抗病毒药的模型：与这种方法紧密对准的是使用PFS和其他病毒的petides的潜力来治疗未满足的医疗需求通过导致有害并发症的感染。