Novel & Selective Small Molecular Inhibitors of Human Peptide Deformylase

描述（由申请人提供）：本研究的目标是开发一种适用于高通量筛选化学文库的稳健且敏感的检测方法，从而识别人肽去甲酰基酶（HsPDF）的特定抑制剂。在多种肿瘤细胞系中，actionin抑制HsPDF活性和RNA干扰抑制HsPDF表达均可导致增殖阻滞。因此，HsPDF构成了抗癌药物的新靶点，特异性HsPDF抑制剂可能构成一类新的抗肿瘤药物。为了鉴定基于不可能与蛋白酶非特异性相互作用的独特基序的新抑制剂，我们将继续筛选一个包含20万种化合物的大型文库。为了实现这一目标，我们计划开发一种基于两种不同检测方法的高通量筛选策略。主筛选将采用一种新型的荧光偏振分析来快速识别HsPDF结合物，而基于功能分析的二次筛选将进行，以选择最佳的HsPDF抑制剂。这两步筛选策略背后的基本原理是利用快速和稳健的FP测定，在初级筛选中有效地筛选出与HsPDF相互作用的化合物。这将使我们能够将更耗时的基于PDF酶活性的分析应用于数量大大减少的化合物，以确认命中。结合使用这两种测定法的另一个优点在于其可靠性。基于两种不同技术的筛选可能会导致确定可信的命中。事实上，与仅基于一种技术的策略相比，化合物干扰两种分析并因此在两种分析中表现为假阳性的可能性大大降低。

DESCRIPTION (provided by applicant): The goal of this study is to develop a robust and sensitive assay adaptable to high throughput screening of chemical libraries, which in turn would identify specific inhibitors of human peptide deformylase (HsPDF). Both the inhibition of HsPDF activity by actinonin, and the inhibition of HsPDF expression by RNA interference lead to proliferation arrest in a variety of tumor cell lines. HsPDF therefore constitutes a novel target for anticancer agents, and specific HsPDF inhibitors could constitute a new class of antitumor agents. For the purpose of identifying new inhibitors based on unique motifs not likely to interact non-specifically with proteases, we will proceed to the screening of a large library of 200,000 compounds. To achieve that goal, we plan to develop a high-throughput screening strategy based on two different assays. A primary screen will employ a novel fluorescence polarization assay to rapidly identify HsPDF binders, and a secondary screen based on a functional assay will be performed in order to select the best HsPDF inhibitors. The rationale behind this two-step screening strategy is to efficiently sort out compounds interacting with HsPDF in a primary screen using the quick and robust FP assay. This will allow us to apply the more time-consuming assay based on PDF enzymatic activity only to a vastly reduced number of compounds, in order to confirm the hits. Another advantage in using this combination of two assays lies in its reliability. A screen based on two different techniques is likely to lead to the identification of trustable hits. Indeed, the likelihood that a compound interferes with both assays, and therefore behaves as a false positive in both assays, is vastly reduced compared to a strategy solely based on one technique.