NOX1 and NOX2 as Therapeutic Targets in Influenza

NOX1 和 NOX2 作为流感的治疗靶点

• 批准号: 8490301
• 负责人: John David Lambeth
• 金额: $ 22.54万
• 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 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8490301
• 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; 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

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.

描述（由申请方提供）：流感病毒显示出高突变率和重组率，很快使免疫无效，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可用抗病毒分子具有抗性。这些因素证明了对靶向宿主编码功能的药物的未满足的医疗需求，因此不受病毒选择的影响。此外，偶尔会出现高致死性流感病毒株（例如1918年流感、禽流感），通过这些病毒诱导介导肺细胞功能障碍和损伤（急性呼吸窘迫综合征或ARDS）的“细胞因子风暴”的倾向而导致发病率/死亡率。这种致病性变化包括肺泡液体转运的破坏、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，不存在预防肺损伤的有效治疗方法。其他病原体也发生了类似的变化，包括SARS冠状病毒和炭疽，针对流感开发的宿主导向疗法预计也会有效。 Nox酶是产生超氧化物和次级活性氧（ROS）的NADPH氧化酶，其充当信号分子，并且在高浓度下直接破坏生物分子。我们提出了一个涉及上皮细胞NOX 1和单核细胞/中性粒细胞NOX 2产生的ROS的信号级联反应作为关键步骤：a）促进病毒复制和/或传播和B）介导肺损伤。Nox 1产生的上皮细胞活性氧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敲除小鼠和抑制剂，初步证据表明，抑制Nox 1和Nox 2将在流感感染中具有治疗益处。该应用程序代表了埃默里大学和疾病控制中心（CDC）之间的合作努力，在那里可以进行使用高毒力流感病毒株（PR 8，H5 N1禽流感）的研究。兰贝斯实验室以其在氮氧化物发现、酶学和细胞生物学方面的专业知识而闻名，已经发现（使用高通量和低通量筛选）四种化学系列的小分子氮氧化物抑制剂。与埃默里药物发现研究所合作，我们的团队将进一步开发这些抑制剂，提高其效力，异构体选择性，代谢稳定性和药理学特性，并将协调临床前开发作为候选药物。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缺失的小鼠中，以证明Nox 1和/或Nox 2作为治疗靶点的概念验证，2）在感染和未感染的WT小鼠中测试候选药物。总体目标是开发靶向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/传播的宿主信号传导途径。此类化合物通常可用于治疗导致严重肺功能障碍/病理的所有流感病毒株和可能的其它病原体（SARS-CoV、炭疽）。

项目成果

NADPH Oxidase 1 Is Associated with Altered Host Survival and T Cell Phenotypes after Influenza A Virus Infection in Mice.

• DOI: 10.1371/journal.pone.0149864
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Hofstetter AR;De La Cruz JA;Cao W;Patel J;Belser JA;McCoy J;Liepkalns JS;Amoah S;Cheng G;Ranjan P;Diebold BA;Shieh WJ;Zaki S;Katz JM;Sambhara S;Lambeth JD;Gangappa S
• 通讯作者: Gangappa S