Core A - Discovery Core

核心 A - 发现核心

基本信息

批准号：
10465059
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/10465059
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 Drug Targeting Ensure Enzymes Evaluation Fluorescence-Activated Cell Sorting G-Protein-Coupled Receptors GPR68 gene Genomics Human Infrastructure Institutes Knowledge Lead Libraries Ligands Modeling Molecular Muscarinic Acetylcholine Receptor Muscarinic Acetylcholine Receptor M3 Muscle Contraction Natural Products Pathway interactions Peptide Library Pharmaceutical Chemistry 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 Vestibule airway inflammation base constriction design effective therapy extracellular high throughput screening improved inhibitor innovation lead candidate lead optimization novel predictive test respiratory smooth muscle screening small molecule small molecule inhibitor success 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.

项目概要筛选核心将建立在我们利用传统和虚拟筛选方法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整合。工作流程使我们能够快速从初步筛选结果转向详细的结构-活性关系（SAR），以及从计算预测到定量实验数据。与其他区域筛选资源建立（兰克瑙化学基因组中心（LCGC）；天然产品发现研究所 (NPDI)；莫内尔化学感官中心 (MCSC)）确保获得拟议的筛选项目的高质量分子多样性，以及我们在第二方面经过验证的专业知识信使和蛋白质相互作用测定将加快发现偏向信号传导的探针和靶点使用排列和表达的分子多样性来源研究气道平滑肌中的通路。 Jefferson Discovery Core 为 Core A 提供传统筛选功能，使用 in- 拥有分子多样性来源以及从区域和国家资源中获得的图书馆（NCI- DTP）来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) 检测（cAMP、Ca2+）和蛋白质相互作用检测。（酶片段互补）已针对多孔板和通过检测进行了优化荧光激活细胞分选，可以使用阵列和表达的分子来源用于鉴定调节影响支气管运动张力的信号通路的探针和靶标的多样性。 Gα12 信号传导抑制剂的 HTS（项目 1）、TAS2R14 的内化（项目 2）、HTS、SAR 和药物化学发现和优化 2AR 的 Gs 偏向激动剂和变构调节剂（项目 3）和通过虚拟筛选（项目 4）预测的 OGR1 的 Gs 偏向变构调节剂的评估是最重要的优先事项。核心 A 使用计算建模和对接方法来指导和提供信息基于 CHARMM 的传统筛选方法将用于预测。通过高通量筛选确定小分子的结合模式，开发 SAR 模型并预测具有改善的活性和理化性质的衍生物。平行的 HTS 实验工作，特别是对 HTS 中确定的最有希望的命中进行跟进。 2AR 和 OGR1 小分子变构调节剂的并行建模研究将针对将使用基于结构的知识来识别导致 Gs 偏向信号传导的常见结构特征。为并行虚拟筛选策略提供信息，以识别 2AR 和 OGR1 的新型变构调节剂。虚拟筛选方法将用于识别 G12 和 Gq 的小分子抑制剂，该抑制剂可模拟 Gq 特异性抑制剂 YM-254890 的药理作用。