Inhibition of s-nitrosoglutathione reductase for the treatment of asthma

描述（由申请人提供）：哮喘是一种由呼吸道慢性炎症引起的疾病。发炎的气道对各种吸入刺激反应过度，并因支气管平滑肌收缩而阻塞。哮喘患者和实验动物气道内内源性支气管扩张剂s-亚硝基谷胱甘肽（GSNO）的耗竭提示气道内源性支气管扩张剂s-亚硝基谷胱甘肽（GSNO）的耗竭可能是气道粘膜液中GSNO的耗竭。在GSNO代谢酶s-亚硝基谷胱甘肽还原酶（GSNOR）缺乏的小鼠中进行的研究支持了这一点。缺乏GSNOR的小鼠即使在存在强烈炎症的情况下也不会出现气道高反应性。当前研究的长期目标是设计人类GSNOR抑制剂，并评估预防GSNO分解在哮喘治疗中的有效性。我们将使用高通量筛选方法来鉴定结合人GSNOR的化合物。我们将利用酶动力学、x射线晶体学和细胞培养模型中蛋白质s-亚硝基化水平变化的测量，进一步评估鉴定的化合物作为研究GSNOR抑制对气道高反应性影响的线索的潜力。拟议的工作将导致鉴定化合物，这将允许评估预防哮喘患者哮喘发作的新方法。

DESCRIPTION (provided by applicant): Asthma is a disorder arising from chronic inflammation of the airways. The inflamed airways become hyper-responsive to various inhaled stimuli and become obstructed as result of contraction of bronchial smooth muscles. Depletion of the endogenous bronchodilator, s-nitrosoglutathione (GSNO) from the airways of asthmatics and experimental animals suggested that one of the underlying causes of airflow obstruction to be the loss of GSNO from the airway lining fluid. This has been supported by studies in mice deficient in GSNO metabolizing enzyme, s-nitrosoglutathione reductase (GSNOR). Mice deficient in GSNOR were protected from airway hyper-responsivity even in presence of strong inflammation. The long term objective of the current study is to design inhibitors of human GSNOR and evaluate the usefulness of preventing GSNO breakdown in the treatment of asthma. We will use high throughput screening methods to identify compounds that bind human GSNOR. We will further evaluate the potential of the identified compounds as leads for investigating the effects of GSNOR inhibition on airway hyper-responsivity using enzyme kinetics, x-ray crystallography and measurement of changes in the protein s-nitrosylation levels in a cell culture model. The proposed work will lead to identification of compounds that will allow evaluating a new approach for preventing asthmatic episodes in asthma patients.