Novel & Selective Small Molecular Inhibitors of Human Peptide Deformylase

描述(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可靠和灵敏的检测方法，进而识别特定的人肽变形酶抑制物(HsPDF)。放线菌素对HsPDF活性的抑制和RNA干扰对HsPDF表达的抑制均导致多种肿瘤细胞的增殖停滞。因此，HsPDF构成了抗癌药物的新靶点，而特定的HsPDF抑制剂可能构成一类新的抗肿瘤药物。为了根据不太可能与蛋白酶非特异性相互作用的独特基序识别新的抑制剂，我们将继续筛选包含200,000个化合物的大型文库。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。一次筛选将采用一种新的荧光偏振分析来快速鉴定HsPDF结合物，二次筛选将基于功能分析进行，以选择最好的HsPDF抑制剂。这种两步筛选策略背后的基本原理是使用快速而可靠的FP分析在初级筛查中有效地筛选出与HsPDF相互作用的化合物。这将使我们能够将基于PDF酶活性的更耗时的分析仅应用于数量大大减少的化合物，以确认命中。使用这两种检测方法结合使用的另一个优势在于其可靠性。基于两种不同技术的屏幕可能会导致识别可信的命中。事实上，与仅基于一种技术的策略相比，一种化合物干扰这两种检测并因此在两种检测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.