Chemogenomic identification of Ref-l/AP-1 as a therapeutic target for asthma
Chemogenomic identification of Ref-l/AP-1 as a therapeutic target for asthma
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DOI：
10.1073/pnas.0437889100
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发表时间：
2003-02-04
影响因子：
11.1
通讯作者：
Kahn, M
中科院分区：
文献类型：
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作者：
Nguyen, C;Teo, JL;Kahn, M
哮喘的特征在于肺中的氧化剂/抗氧化剂失衡，导致氧化还原敏感性转录因子、核因子κ B（NF-κ B）和激活蛋白-1（AP-11）的激活。为了开发哮喘的治疗策略，我们使用化学基因组学方法来筛选AP-1转录的小分子抑制剂。我们开发了一种β链模拟模板，作为氧化还原蛋白的可逆抑制剂（假底物）。该模板结合了烯二酮部分，以在氧化还原蛋白的活性位点中捕获反应性半胱氨酸亲核试剂。通过使用组合库方法，通过X和Y官能度的变化来实现单个氧化还原因子的特异性。构建有限阵列（2 × 6），其中X是NHCH 3或NHCH 2 Ph，Y是甲基、苯基、间氰基苯基、间硝基苯基、间乙酰基苯胺或间甲基苯甲酸酯。评价这些类似物在瞬时转染的人肺上皮A549细胞中抑制来自AP-1或NF-κ B报告基因的转录的能力。鉴定了一种小分子抑制剂PNRI-299，其选择性抑制AP-1转录（IC 50为20 μ M）而不影响NF-κ B转录（高达200 μ M）或硫氧还蛋白（高达200 μ M）。通过亲和层析方法确定PNRI-299的分子靶标是氧化还原酶，氧化还原效应因子-1。在小鼠哮喘模型中，选择性氧化还原效应因子-1抑制剂PNRI-299显著降低气道嗜酸性粒细胞浸润、粘液分泌过多、水肿和IL-4水平。这些数据证实AP-1作为过敏性气道炎症的重要治疗靶点。
Asthma is characterized by an oxidant/antioxidant imbalance in the lungs leading to activation of redox-sensitive transcription factors, nuclear factor kappaB (NF-kappaB), and activator protein-1 (AP-11). To develop therapeutic strategies for asthma, we used a chemogenomics approach to screen for small molecule inhibitor(s) of AP-1 transcription. We developed a beta-strand mimetic template that acts as a reversible inhibitor (pseudosubstrate) of redox proteins. This template incorporates an enedione moiety to trap reactive cysteine nucleophiles in the active sites of redox proteins. Specificity for individual redox factors was achieved through variations in X and Y functionality by using a combinatorial library approach. A limited array (2 x 6) was constructed where X was either NHCH3 or NHCH2 Ph and Y was methyl, phenyl, m-cyanophenyl, m-nitrophenyl, m-acetylaniline, or m-methylbenzoate. These analogs were evaluated for their ability to inhibit transcription in transiently transfected human lung epithelial A549 cells from either an AP-1 or NF-kappaB reporter. A small-molecule inhibitor, PNRI-299, was identified that selectively inhibited AP-1 transcription (IC50 of 20 muM) without affecting NF-kappaB transcription (up to 200 muM) or thioredoxin (up to 200 muM). The molecular target of PNRI-299 was determined to be the oxidoreductase, redox effector factor-1 by an affinity chromatography approach. The selective redox effector factor-1 inhibitor, PNRI-299, significantly reduced airway eosinophil infiltration, mucus hypersecretion, edema, and IL-4 levels in a mouse asthma model. These data validate AP-1 as an important therapeutic target in allergic airway inflammation.