Screening for inhibitors of allergen-associated airway smooth muscle contraction

项目总结 尽管使用大剂量吸入皮质类固醇加支气管扩张剂进行积极治疗，但约有5%-10% 大多数哮喘患者(1500万至3000万)出现严重的危及生命的支气管收缩症状。 在这些严重哮喘患者中，有近一半的人也对普通哮喘“致敏”(即有IgE介导的过敏反应)。 霉菌，如烟曲霉(Af)。这种情况现在已经被认为是一个不同的实体， 指定为真菌性哮喘(FA)。不幸的是，目前对FA的治疗-抗真菌药和/或奥马珠单抗(抗- IGE)--从长远来看，并没有被证明是有效的。 通过研究Af诱导的小鼠和哮喘患者的支气管收缩，我们的团队发现了一种新的， FA中的直接和可药物靶标-Af过敏原衍生的丝氨酸蛋白酶，碱性蛋白酶1(Alp1)。Alp1 通过诱导呼吸道平滑肌(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。在此应用程序中，我们 将继续提出这样的假设，即Alp1诱导的人类ASM收缩的抑制剂可以通过高密度脂蛋白 吞吐量筛选(HTS)。在目标1中，我们将使用HTS来鉴定Alp1蛋白酶的小分子抑制剂。在……里面 目的2，探讨FA病理生物学对Alp1抑制剂治疗效果的影响。在《目标3》中，我们将 确定Alp1抑制物治疗的机制及检测意义。我们希望我们的方法能够确定 新的抗FA药物候选药物，通过这样做，提供了对ASM固有和过敏原的实质性洞察 蛋白水解酶依赖于支气管收缩的机制。

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.