Core A - Discovery Core

核心 A - 发现核心

基本信息

批准号：
10683121
负责人：
Jeffrey L Benovic
金额：
$ 29.46万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10683121
关键词：
Adrenergic Receptor Affect Affinity Agonist Airway Disease Arrestins Asthma Back Binding Biochemical Biological Assay Bronchodilation Cells Chemicals Complement Complex Computer Models Computing Methodologies Cyclic AMP Data Detection Development Disease Docking Ensure Enzymes Evaluation Fluorescence-Activated Cell Sorting G-Protein-Coupled Receptors GPR68 gene Genomics Human Infrastructure Knowledge Lead Libraries Ligands Modeling Molecular Muscarinic Acetylcholine Receptor Muscarinic Acetylcholine Receptor M3 Muscle Contraction Natural Products Pathway interactions Peptide Library Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacologic Actions Phenotype Play Population Private Sector Property Proteins Public Sector Quinazolines Relaxation Resources Role Second Messenger Systems Signal Pathway Signal Transduction Source Specificity Structure Structure-Activity Relationship Study models Surface Plasmon Resonance Testing Vestibule airway inflammation constriction design effective therapy extracellular high throughput screening improved inhibitor innovation lead candidate lead optimization novel respiratory smooth muscle screening small molecule small molecule inhibitor success synergism translational applications virtual screening

项目摘要

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.

项目摘要筛选核心将利用传统和虚拟筛选方法的成功建立在我们的成功基础上识别调节支气管运动张力的小分子。传统和虚拟筛选的集成工作流程使我们能够从主要筛选命中迅速移动到详细的结构活动关系（SAR），从计算预测到定量实验数据。核心的关系已与其他区域筛查资源（Lankenau Chemical Genomics Center（LCGC）建立；天然产品发现研究所（NPDI）； Monell化学感官中心（MCSC））确保进入拟议筛查项目的高质量分子多样性，以及我们在第二次验证的专业知识 Messenger和蛋白质相互作用测定将加快发现偏向信号传导的问题和目标使用阵列和表达的分子多样性来源，气道平滑肌的途径。 Jefferson Discovery Core为Core A提供了常规的筛选功能，并使用In- 分子多样性的房屋来源以及从区域和国家资源获得的图书馆（NCI- dtp）识别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）测定法。第二使者（CAMP，CA2+）和蛋白质相互作用测定法（酶 - 碎片完成）已优化用于在多层板和通过荧光激活的细胞分选，可以使用阵列和表达的分子来源识别问题和目标的多样性，以调节影响支气管计音调的信号通路。 G12信号抑制剂的HTS（项目1），TAS2R14的内在化（项目2），HTS，SAR和医疗化学以发现和优化2AR（项目3）和对虚拟筛选预测的OGR1的GS偏置变构调节剂的评估（项目4）是顶部优先事项。核心A使用计算建模和对接方法来指导和告知常规筛选方法。基于CHARMM的分子对接方法将用于预测通过高通量筛选确定的小分子的结合模式，开发SAR模型和预测具有改善活性和生理特性的衍生物。这些研究与平行的实验HTS工作，特别是在跟踪HTS确定的最有承诺的命中时。 2AR和OGR1的小分子变构调节剂的平行建模研究将针对识别导致GS偏置信号传导的常见结构特征。将使用基于结构的知识告知并行虚拟筛选策略，以识别2AR和OGR1的新型变构调节剂。最后，虚拟筛选方法将用于识别G12和GQ的小分子抑制剂，以模仿 GQ特异性抑制剂YM-254890的药物作用。