Core A - Discovery Core

项目摘要 筛选核心将建立在我们的成功利用传统和虚拟筛选方法， 确定调节支气管张力的小分子。传统和虚拟筛选的整合 工作流程使我们能够从初步筛选命中快速移动到详细的结构-活性关系 (SAR)，以及从计算预测到定量实验数据。核心的关系 与其他区域筛选资源建立了（Lankenau化学基因组学中心（LCGC）; 天然产品发现研究所（NPDI）;蒙乃尔化学感官中心（MCSC））确保获得 高质量的分子多样性，为拟议的筛选项目，以及我们在第二个验证的专业知识， 信使和蛋白质相互作用分析将加快发现探针和目标， 使用排列和表达的分子多样性来源在气道平滑肌中的通路。 Jefferson Discovery Core为Core A提供常规筛选能力，使用- 容纳分子多样性来源以及从区域和国家资源获得的库（NCI- DTP）来鉴定产生与基于细胞的气道松弛相关的表型的小分子， 高通量筛选（HTS）测定。第二信使（cAMP，Ca 2+）和蛋白质相互作用测定 （酶片段互补）已被优化，用于在多孔板中检测， 荧光激活的细胞分选，其使得能够使用阵列的和表达的分子源， 用于鉴定调节影响支气管张力的信号传导途径的探针和靶标的多样性。 HTS用于G β 12信号传导抑制剂（项目1）、TAS 2 R14内化（项目2）、HTS、SAR和药物 化学发现和优化Gs-biased激动剂和变构调节剂的α 2 AR（项目3）， 通过虚拟筛选（项目4）预测的OGR 1的GS偏置变构调节剂的评价是最高的 优先事项 核心A使用计算建模和对接方法来指导和通知 传统的筛选方法。基于CHARMM的分子对接方法将用于预测 从高通量筛选中识别的小分子的结合模式，开发SAR模型， 预测具有改进的活性和物理化学性质的衍生物。这些研究是协同作用， 平行的实验性高温超导努力，特别是在后续的最有希望的命中确定高温超导。 平行建模研究的小分子变构调节剂的β 2 AR和OGR 1将针对 识别导致GS偏置信号传导的共同结构特征。将使用基于结构的知识 为并行虚拟筛选策略提供信息，以鉴定新型的β 2 AR和OGR 1的变构调节剂。最后， 一种虚拟筛选方法将用于鉴定G12和Gq的小分子抑制剂， Gq特异性抑制剂YM-254890的药理作用。

Project Summary The screening core will build on our success utilizing conventional and virtual screening approaches to identify small molecules that regulate bronchomotor tone. The integration of conventional and virtual screening workflows enabled us to move rapidly from primary screening hits to detailed structure-activity relationships (SAR), and from computational predictions to quantitative experimental data. The relationships that the core has established with other regional screening resources (Lankenau Chemical Genomics Center (LCGC); Natural Products Discovery Institute (NPDI); Monell Chemical Senses Center (MCSC)) ensures access to high-quality molecular diversity for proposed screening projects, and our validated expertise in second messenger and protein interaction assays will expedite discovery of probes and targets that bias signaling pathways in airway smooth muscle using arrayed and expressed sources of molecular diversity. The Jefferson Discovery Core provides the conventional screening capabilities for Core A, using in- house sources of molecular diversity as well as libraries obtained from regional and national resources (NCI- DTP) to identify small molecules engendering phenotypes that correlate with airway relaxation in cell-based, high-throughput screening (HTS) assays. Second messenger (cAMP, Ca2+) and protein interaction assays (enzyme-fragment complementation) have been optimized for detection both in multiwell plates and by fluorescence-activated cell sorting, which enables use of both arrayed and expressed sources of molecular diversity for identification of probes and targets regulating signaling pathways affecting bronchomotor tone. HTS for inhibitors of G12 signaling (project 1), internalization of TAS2R14 (project 2), HTS, SAR and medicinal chemistry to discover and optimize Gs-biased agonists and allosteric modulators of 2AR (project 3) and evaluation of Gs-biased allosteric modulators of OGR1 predicted by virtual screening (project 4) are the top priorities. Computational modeling and docking approaches are used by Core A both to direct and inform conventional screening approaches. CHARMM-based molecular docking approaches will be used to predict the binding modes of small-molecules identified from high-throughput screening, develop SAR models and predict derivatives with improved activity and physiochemical properties. These studies are synergistic with parallel experimental HTS efforts, particularly in following-up on the most promising hits identified from HTS. Parallel modeling studies of small-molecule allosteric modulators of 2AR and OGR1 will be aimed at identifying common structural features leading to Gs-biased signaling. Structure-based knowledge will be used to inform parallel virtual screening strategies to identify novel allosteric modulators of 2AR and OGR1. Finally, a virtual screening approach will be used to identify small-molecule inhibitors of G12 and Gq that would mimic the pharmacological action of the Gq specific inhibitor YM-254890.