Developing a New Antioxidant Activator to Intervene in Lung Inflammatory Diseases

开发一种新型抗氧化剂激活剂来干预肺部炎症疾病

• 批准号: 7822223
• 负责人: John A Willimann
• 金额: $ 16.39万
• 依托单位: GLOBAL LIFE TECHNOLOGIES CORPORATION
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7822223
• 关键词: Acute; Acute Lung Injury; Allergens; American; Animal Model; Antioxidants; Attention; Bacterial Infections; Binding; Biological; Biological Models; Cell Culture Techniques; Cell Nucleus; Cells; Chemicals; Chronic; Chronic Obstructive Airway Disease; Chronic lung disease; Clinical Trials; Cytoprotection; DNA Binding; Dependence; Development; Disease; Drug Kinetics; EMSA; Effectiveness; Endotoxins; Enzymes; Extrinsic asthma; Gene Activation; Gene Expression; Gene Proteins; Gene Targeting; Genes; Goals; Health; Host Defense; Human; Hypersensitivity; Immune response; Inflammation; Inflammatory; Intervention; Kinetics; Laboratories; Lead; Life; Link; Lung; Lung diseases; Measures; Methodology; Microarray Analysis; Modeling; Monitor; Morbidity - disease rate; Mus; Nasal Epithelium; Nuclear Protein; Nuclear Proteins; Oils; Outcome; Ovalbumin; Oxidants; Oxidative Stress; Ozone; Pathologic Processes; Pathology; Pathway interactions; Phase; Phase I Clinical Trials; Play; Pneumonia; Preclinical Testing; Preparation; Preventive; Pulmonary Pathology; Relative (related person); Reporter; Research; Research Personnel; Respiratory System; Reverse Transcriptase Polymerase Chain Reaction; Role; Safety; Screening procedure; Sepsis; Signal Transduction; Stimulus; Stream; Structure of parenchyma of lung; Testing; Therapeutic Agents; Time; Tissues; Toxic effect; Viral; Volatile Oils; antimicrobial; base; cell type; cytotoxicity; human disease; human subject; in vivo; inflammatory marker; mouse model; novel; oxidant stress; promoter; protein expression; public health relevance; research and development; research clinical testing; respiratory; response; transcription factor

DESCRIPTION (provided by applicant): Pulmonary inflammatory diseases linked to oxidant stress in lung tissues represent a major health concern in the U.S. These diseases include allergic asthma, chronic obstructive pulmonary disease, viral and bacterial infections, and related sepsis-induced acute lung injury. Evidence indicates that antioxidant defenses in the tissues play a pivotal role in reducing pulmonary inflammation and that enhancement of these protective pathways can lead to a reduction of tissue damage and morbidity. Emerging evidence has identified a central role for the transcription factor, Nrf2, in orchestrating the induction of a global network of tissue-protective antioxidant enzymes in response to oxidant-related stimuli. Much attention is being focused on means by which to stimulate activation of Nrf2 for both preventive and interventive therapies. Our preliminary studies have demonstrated that a constituent of a natural oil induces marked activation of Nrf2 and subsequent stimulation of its down-stream antioxidant gene targets. The Objectives of the Phase I research are to test the effectiveness and safety of the oil containing the active constituent in order to determine its potential for continued development in Phase II R&D as a therapeutic agent. Phase I Specific Aims are to: (1) Identify the component oil of the previously-tested oil mixture in which the primary Nrf2-stimulating activity resides. This will be achieved using cells stably transfected with an Nrf2 reporter construct followed by verification of antioxidant gene and protein expression in wild-type cells. (2) Determine the pharmacokinetics and Nrf2-selectivity of the active oil. The time-course of expression and efficacy of stimulation of antioxidant and cytotoxicity marker genes relative to known Nrf2 chemical activators will be examined. Nrf2 signaling will be assessed by nuclear protein accumulation and DNA binding (EMSA) and the absence of activity in Nrf2 -/- cells. Microarray analysis will identify the presence of concurrently-activated pathways. (3) Assess the effectiveness of the active constituent in reducing markers of inflammation and tissue damage in mouse models of allergic asthma and endotoxin- induced acute lung injury. Markers will include histopathologic measures and the expression of genes and proteins associated with the oxidant-related inflammatory pathology of these models. These studies will be carried out using cell culture and animal model systems and methodologies currently in use by the investigators. The activities of Phase I are expected to provide the scientific basis for further isolation and development of the active constituent present in the natural oil. It is anticipated that the outcome of Phase I and II research and development will be the identification of a molecule that will be attractive as a commercial product ready for development as a therapeutic agent. PUBLIC HEALTH RELEVANCE: Over 35 million Americans live with acute and chronic lung diseases whose pathology is linked to inflammation. Antioxidants synthesized in lung tissues are known to offer protection against pathologic processes associated with many of these diseases. The goal of this project is to isolate and test the effectiveness of an agent found in an essential oil that has been demonstrated to activate broad synthesis of protective antioxidants in the lung.

描述（由申请人提供）：与肺组织中与氧化剂胁迫有关的肺部炎症性疾病代表了美国的主要健康问题，这些疾病包括过敏性哮喘，慢性阻塞性肺部疾病，病毒和细菌感染以及相关的SEPSIS诱导的急性急性肺损伤。证据表明，组织中的抗氧化剂防御在减少肺部炎症中起着关键作用，并且这些保护途径的增强可以导致组织损伤和发病率的降低。新兴的证据已经确定了转录因子NRF2在策划响应与氧化剂相关刺激的全球组织保护抗氧化剂酶网络中的核心作用。大量关注的重点是用于预防疗法和干预疗法的NRF2激活的手段。我们的初步研究表明，天然油的组成诱导了NRF2的明显激活以及随后刺激其下游抗氧化剂基因靶标。第一阶段研究的目标是测试包含活性成分的石油的有效性和安全性，以确定其作为治疗剂的II阶段R＆D持续发展的潜力。第一阶段的特定目的是：（1）确定原发性NRF2刺激活性所在的先前测试的油混合物的成分油。这将使用稳定转染的NRF2报告基因构建体稳定转染的细胞，然后验证野生型细胞中抗氧化基因和蛋白质表达。 （2）确定活性油的药代动力学和NRF2 s-sectitive。将研究相对于已知NRF2化学激活剂的抗氧化剂和细胞毒性标记基因刺激的表达时间和疗效。 NRF2信号传导将通过核蛋白积累和DNA结合（EMSA）以及NRF2 - / - 细胞中没有活性来评估。微阵列分析将确定同时激活的途径的存在。 （3）评估活性成分在减少过敏性哮喘和内毒素诱导的急性肺损伤的小鼠模型中降低炎症和组织损伤标记的有效性。标记将包括组织病理学测量以及与这些模型的氧化剂相关炎症病理学相关的基因和蛋白质的表达。这些研究将使用研究人员目前正在使用的细胞培养和动物模型系统和方法进行。 I期的活动有望为天然石油中存在的活跃成分的进一步隔离和发展提供科学基础。可以预料，第一阶段和第二阶段的研发结果将是鉴定一个分子，该分子将作为准备作为治疗剂开发的商业产品具有吸引力。公共卫生相关性：超过3500万美国人患有与炎症有关的急性和慢性肺部疾病。已知在肺组织中合成的抗氧化剂可为与许多此类疾病相关的病理过程提供保护。该项目的目的是隔离和测试精油中发现的药物的有效性，该药油已被证明激活了肺中保护性抗氧化剂的广泛合成。