Screening for inhibitors of allergen-associated airway smooth muscle contraction

项目摘要 尽管使用大剂量吸入性皮质类固醇加支气管扩张剂进行了积极治疗， 的哮喘患者（1500 - 3000万）经历严重和危及生命的支气管收缩症状。 这些严重哮喘患者中有近一半还对常见的过敏原“敏感”（即具有IgE介导的过敏）。 霉菌如烟曲霉（Af）。这种情况现在已被确认为一个独特的实体， 真菌性哮喘（FA）不幸的是，FA-抗真菌剂和/或奥马珠单抗（抗真菌药）的当前疗法是不理想的。 IgE）-尚未证明长期有效。 通过研究小鼠和哮喘患者中AF诱导的支气管收缩，我们的团队发现了一种新的， FA中的直接、可药用靶标-Af变应原衍生的丝氨酸蛋白酶，碱性蛋白酶1（Alp 1）。Alp1 通过诱导气道平滑肌（ASM）促进FA的定义症状-支气管收缩 收缩，通过似乎独立于过敏性炎症的机制。在本申请中，我们 将继续研究Alp 1诱导的人ASM收缩抑制剂可以通过高通量的方法发现的假设。 通量筛选（HTS）。在目标1中，我们将使用HTS来鉴定Alp 1蛋白酶的小分子抑制剂。在 目的2，我们将检查FA病理生物学对Alp 1抑制剂治疗效果的影响。在目标3中，我们 确定Alp 1抑制剂治疗的机制和意义。我们希望我们的方法能够识别 新的抗FA候选药物，并在这样做，提供了实质性的洞察ASM的内在和过敏原 支气管收缩的蛋白酶依赖机制。

PROJECT SUMMARY Despite aggressive treatment with high-dose inhaled corticosteroids plus bronchodilators, approximately 5-10% of people with asthma (15-30 million) experience severe and life-threatening symptoms of bronchoconstriction. Nearly one-half of these severe asthma patients are also “sensitized” (i.e. have IgE mediated allergy) to common molds such as Aspergillus fumigatus (Af). This condition has now been recognized as a distinct entity and designated fungal asthma (FA). Unfortunately, current therapies for FA—antifungals and/or omalizumab (anti- IgE)—have not proven to be efficacious in the long-term. By studying Af-induced bronchoconstriction in mice and people with asthma, our team has discovered a novel, direct, and druggable target in FA—the Af allergen-derived serine protease, alkaline protease 1 (Alp1). Alp1 promotes the defining symptom of FA—bronchoconstriction—by inducing airway smooth muscle (ASM) contraction, through mechanisms that appear to be independent of allergic inflammation. In this application, we will pursue the hypothesis that inhibitors of Alp1-induced human ASM contraction can be discovered by high throughput screening (HTS). In aim 1, we will use HTS to identify small molecular inhibitors of Alp1 protease. In aim 2, we will examine the effects of FA pathobiology on the efficacy of Alp1-inhibitor therapy. In aim 3, we will determine mechanism and examine significance for Alp1-inhibitor therapy. We expect our approach to identify novel anti-FA drug candidates, and in doing so, offer substantial insight into both ASM-intrinsic and allergen protease dependent mechanisms of bronchoconstriction.