NOX1 and NOX2 as Therapeutic Targets in Influenza

NOX1 和 NOX2 作为流感的治疗靶点

• 批准号: 8889190
• 负责人: John David Lambeth
• 金额: $ 45.96万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8889190
• 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 Kinetics; Drug Targeting; 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; Severe Acute Respiratory Syndrome; 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; small molecule; superoxide-generating NADPH oxidase; therapeutic target; therapy development

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 targeted 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/mortality 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, and 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-Coronovirus 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 further 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.

流感病毒表现出很高的变异率和重组率，很快就会使免疫无效和 需要每年生产疫苗。此外，高达85%的分离株对现有的抗病毒药物具有抗药性。 针对病毒本身的分子。这些因素表明，对药物的医疗需求尚未得到满足 目标是宿主编码的功能，因此不受病毒选择的影响。此外，高度致命的 流感病毒株(如1918年流感、禽流感)偶尔会出现，通过 这些病毒诱发“细胞因子风暴”的倾向，导致肺细胞功能障碍和损害(急性 呼吸窘迫综合征或ARDS)。这种病原性变化包括肺泡液的破坏。 中性粒细胞和中性粒细胞对肺泡上皮细胞的转运、凋亡和肺组织的浸润/破坏 单核细胞。目前，尚不存在预防肺损伤的有效治疗方法。类似的变化也发生在 其他病原体包括SARS冠状病毒和炭疽热，在这些地方，宿主指导的治疗方法被开发出来 预计流感也会有效。 NOx酶是一种NADPH-氧化酶，能产生超氧化物和次生活性氧物种 (ROS)作为信号分子，在高浓度下直接损害生物分子。我们建议 上皮细胞NOX1和单核细胞/PMN NOX2产生的ROS作为关键步骤的信号级联反应 那就是：a)促进病毒复制和/或传播，b)调节肺损伤。上皮性ROS的产生 By Nox1是最早触发细胞因子风暴和细胞功能变化的事件之一。使用NOX 基因敲除小鼠和抑制剂，初步证据表明，NOX1和NOX2的抑制将是 对流感感染有治疗作用。这个应用程序代表了 埃默里和疾病控制中心(CDC)，在那里使用高毒力流感毒株进行研究 (PR8，H5N1禽流感)可以进行。兰贝斯实验室以其在氮氧化物发现方面的专业知识而闻名， 酶学和细胞生物学，已经发现(使用高通量和低通量筛选)四种化学系列 小分子氮氧化物抑制剂。在与埃默里药物发现研究所的合作下，我们的小组将 进一步开发这些抑制剂，提高它们的效力、异构体选择性、代谢稳定性和 药理特性，并将作为候选药物协调临床前开发。恒河 CDC流感小组的一部分实验室将：1)将活病毒感染到基因缺失的小鼠中，以 展示NOX1和/或NOX2作为治疗靶点的概念验证，以及2)测试候选药物 感染和未感染WT小鼠。总体目标是开发针对以下目标的新型临床前候选药物 NOx产生的ROS，从而阻断导致肺组织损伤和病毒感染的宿主信号通路 复制/传播。这类化合物一般可用于治疗所有流感病毒株和 可能是导致严重肺功能障碍/病理的其他病原体(SARS冠状病毒、炭疽病)。