Novel & Selective Small Molecular Inhibitors of Human Peptide Deformylase

小说

基本信息

批准号：
7169304
负责人：
HAKIM DJABALLAH
金额：
$ 21.98万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-06-01 至 2008-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7169304
关键词：
NIH Roadmap Initiative tag chemical registry /resource drug discovery /isolation enzyme activity fluorescence polarization high throughput technology inhibitor /antagonist pharmacology small molecule technology /technique development

项目摘要

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.

描述（由申请人提供）：本研究的目标是开发一种稳健且灵敏的检测方法，适用于化学文库的高通量筛选，进而鉴定人肽去甲酰基酶（HsPDF）的特异性抑制剂。放线菌素对 HsPDF 活性的抑制以及 RNA 干扰对 HsPDF 表达的抑制都会导致多种肿瘤细胞系增殖停滞。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。为了基于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 种化合物的大型库。为了实现这一目标，我们计划开发基于两种不同分析的高通量筛选策略。初级筛选将采用新型荧光偏振测定来快速识别 HsPDF 结合物，并且将进行基于功能测定的二级筛选，以选择最佳的 HsPDF 抑制剂。这种两步筛选策略背后的基本原理是使用快速、稳健的 FP 测定在初级筛选中有效筛选出与 HsPDF 相互作用的化合物。这将使我们能够将基于 PDF 酶活性的更耗时的测定仅应用于大大减少数量的化合物，以确认命中。使用两种检测组合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可信点击的识别。事实上，与仅基于一种技术的策略相比，化合物干扰两种测定并因此在两种测定中表现为假阳性的可能性大大降低。