NOX1 and NOX2 as Therapeutic Targets in Influenza

NOX1 和 NOX2 作为流感的治疗靶点

基本信息

批准号：
8390976
负责人：
John David Lambeth
金额：
$ 19.28万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8390976
关键词：
Acute Adult Respiratory Distress Syndrome Affinity Alveolar Animal Model Animals Anthrax disease Antiviral Agents Antiviral Therapy Apoptosis Appearance Applications Grants Avian Influenza Binding Biodistribution Brain Burn Trauma Canis familiaris Cells Cellular biology Centers for Disease Control and Prevention (U.S.)Chemicals Cherry - dietary Clinical Collaborations Controlled Study Data Development Disease Outbreaks Dose Drug Delivery Systems Drug Kinetics Enzymatic Biochemistry Enzymes Epithelial Epithelial Cells Event Flow Cytometry Functional disorder Generations Genetic Recombination Goals Histology Human Hydration status Immunization In Vitro Infection Infiltration Inflammation Mediators Influenza Influenza A Virus, H5N1 Subtype Institutes Knockout Mice Laboratories Lead Leucocytic infiltrate Life Liquid substance Lung Lung Inflammation Mechanical Ventilators Mediating Medical Metabolic Metabolism Modeling Modification Molecular Morbidity - disease rate Mus Mutation NADPH Oxidase Nox enzyme Nucleoproteins Oxygen Pathogenesis Pathology Pathway interactions Patients Phagocytes Pharmaceutical Chemistry Pharmaceutical Preparations Phase Phospholipids Property Protein Isoforms Protocols documentation Reactive Oxygen Species Resistance SARS coronavirus Series Severities Signal Pathway Signal Transduction Signaling Molecule Solubility Staining method Stains Structure of parenchyma of lung Symptoms Testing Therapeutic Therapeutic Effect Time Tissues Vaccine Production Vaccines Validation Variant Viral Viral Markers Virulent Virus Virus Replication absorption alveolar epithelium base chemokine cytokine design drug candidate drug discovery effective therapy flu improved in vivo influenzavirus inhibitor/antagonist monocyte mortality nephelometry neutrophil novel novel vaccines pandemic disease pandemic influenza pathogen physical property pre-clinical prevent programs public health relevance small molecule superoxide-generating NADPH oxidase therapeutic target therapy development

项目摘要

Description (as provided by the applicant): Influenza virus shows high rates of mutation and recombination that soon renders immunization ineffective and requires yearly production of vaccines. In addition, up to 85% of isolates are resistant to available antiviral molecules targete against the virus itself. These factors demonstrate an unmet medical need for drugs that target host-encoded functions and that are therefore not subject to viral selection. Furthermore, highly lethal strains of influenza (e.g. 1918 flu, bird flu) occasionally arise, causing morbidity/mortaliy through the propensity of these viruses to induce a "cytokine storm" that mediates lung cell dysfunction and damage (acute respiratory distress syndrome or ARDS). Such pathogenic changes include disruptions in alveolar fluid transport, apoptosis of alveolar epithelial cells, an infiltration/destruction of lung tissue by neutrophils and monocytes. Currently, effective treatments to prevent lung damage do not exist. Similar changes occur with other pathogens including SARS-Coronavirus and anthrax, where host-directed therapies developed for influenza are also expected to be effective. Nox enzymes are NADPH-oxidases that generate superoxide and secondary reactive oxygen species (ROS) that act as signaling molecules, and in high concentrations directly damage biomolecules. We propose a signaling cascade involving both epithelial NOX1- and monocyte/PMN Nox2-generated ROS as key steps that: a) facilitate viral replication and/or spread and b) mediate lung damage. The generation of epithelial ROS by Nox1 is among the earliest events that trigger the cytokine storm and cellular functional changes. Using Nox knockout mice and inhibitors, preliminary evidence suggests that inhibition of Nox1 and Nox2 will be therapeutically beneficial in influenza infection. This application represents a collaborative effort between Emory and the Centers for Disease Control (CDC), where studies using highly virulent strains of influenza (PR8, H5N1 bird flu) can be carried out. The Lambeth Laboratory, known for its expertise in Nox discovery, enzymology and cell biology, has discovered (using high- and low-throughput screens) four chemical series of small molecule Nox inhibitors. In collaboration with the Emory Institute for Drug Discovery, our group will furthe develop these inhibitors, improving their potency, isoform selectivity, metabolic stability, and pharmacological properties and will coordinate preclinical development as candidate drugs. The Gangappa Laboratory, part of the Influenza group at the CDC will: 1) infect live virus into genetically deleted mice to demonstrate proof-of-concept for Nox1 and/or Nox2 as therapeutic targets, and 2) test drug candidates in infected and non-infected WT mice. The overall goal is to develop novel pre-clinical drug candidates that target Nox-generated ROS, thereby blocking host signaling pathways that lead to lung tissue damage and viral replication/spread. Such compounds will be generally useful in the treatment of all strains of influenza and likely other pathogens (SARS-CoV, anthrax) that result in severe lung dysfunction/pathology. Public Health Relevance: Highly lethal types of flu and other viruses (e.g. SARS-coronavirus) arise occasionally (bird flu, 1918 flu), and the lack of effective vaccines can result in pandemics that cause large numbers of fatalities. These viruses are lethal because of their tendency to cause a "cytokine storm" resulting in damage to the lung as well as changes in the way the lungs keep fluids from accumulating. The goal of this grant application is to develop and test in animal models compounds that target enzymes in lung cells called NADPH-oxidases (Nox enzymes) and that can be further developed into drugs that inhibit viral replication and treat or prevent the lung damage that occurs in pandemic influenza.

描述（由申请人提供）：流感病毒表现出高突变和重组率，很快就会导致免疫失效，需要每年生产疫苗。此外，高达 85% 的分离株对针对病毒本身的现有抗病毒分子具有耐药性。这些因素表明，对于针对宿主编码功能且因此不受病毒选择影响的药物的医疗需求尚未得到满足。此外，高度致命的流感病毒株（例如 1918 年流感、禽流感）偶尔会出现，这些病毒倾向于诱发介导肺细胞功能障碍和损伤的“细胞因子风暴”，从而导致发病/死亡（急性呼吸窘迫综合征或 ARDS）。这种致病性变化包括肺泡液运输的破坏、肺泡上皮细胞的凋亡、嗜中性粒细胞和单核细胞对肺组织的浸润/破坏。目前，尚不存在预防肺损伤的有效治疗方法。包括 SARS 冠状病毒和炭疽在内的其他病原体也发生了类似的变化，针对流感开发的宿主导向疗法也有望有效。 Nox 酶是 NADPH 氧化酶，可产生超氧化物和次级活性氧 (ROS)，充当信号分子，高浓度时会直接损害生物分子。我们提出涉及上皮 NOX1 和单核细胞/PMN Nox2 生成的 ROS 的信号级联作为关键步骤：a）促进病毒复制和/或传播，b）介导肺损伤。 Nox1 产生的上皮 ROS 是触发细胞因子风暴和细胞功能变化的最早事件之一。使用 Nox 敲除小鼠和抑制剂，初步证据表明，抑制 Nox1 和 Nox2 对流感感染有治疗作用。该应用程序代表了埃默里大学和疾病控制中心 (CDC) 之间的合作成果，可以使用高毒力流感病毒株（PR8、H5N1 禽流感）进行研究。兰贝斯实验室以其在 Nox 发现、酶学和细胞生物学方面的专业知识而闻名，已经（使用高通量和低通量筛选）发现了四种化学系列的小分子 Nox 抑制剂。我们的团队将与埃默里药物发现研究所合作，进一步开发这些抑制剂，提高其效力、亚型选择性、代谢稳定性和药理学特性，并将协调候选药物的临床前开发。 Gangappa 实验室是 CDC 流感小组的一部分，将：1）将活病毒感染到基因删除的小鼠中，以证明 Nox1 和/或 Nox2 作为治疗靶点的概念验证，以及 2）在感染者和非感染者中测试候选药物。 -感染的WT小鼠。总体目标是开发针对 Nox 产生的 ROS 的新型临床前候选药物，从而阻断导致肺组织损伤和病毒复制/传播的宿主信号通路。此类化合物通常可用于治疗所有流感病毒株和可能导致严重肺功能障碍/病理的其他病原体（SARS-CoV、炭疽）。公共卫生相关性：高度致命的流感和其他病毒（例如 SARS 冠状病毒）偶尔会出现（禽流感、1918 年流感），并且缺乏有效的疫苗可能导致大流行导致大量人员死亡。这些病毒之所以致命，是因为它们容易引发“细胞因子风暴”，导致肺部损伤，并改变肺部防止液体积聚的方式。该拨款申请的目标是在动物模型中开发和测试针对肺细胞中称为 NADPH 氧化酶（Nox 酶）的酶的化合物，这些化合物可以进一步开发成抑制病毒复制并治疗或预防发生的肺损伤的药物在大流行性流感中。