Cannabinoid CB2 Receptor Structure and Allosteric Modulators

大麻素 CB2 受体结构和变构调节剂

基本信息

批准号：
10297210
负责人：
Xiang-Qun Xie
金额：
$ 62.4万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-15 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10297210
关键词：
3-Dimensional Affinity Agonist Algorithms Allosteric Site Antibodies Autoimmune Diseases Binding Binding Sites Biological Assay Biology Brain region CNR1 gene CNR2 gene Cannabinoids Chemicals Chronic Complex Cryoelectron Microscopy Crystallization Cyclic AMP Databases Detergents Development Disease Docking Drug Targeting Exhibits Family G-Protein-Coupled Receptors GTP-Binding Proteins Goals Heterogeneity Human Hydrophobicity Immune In Vitro Inflammatory Joints Legal patent Libraries Ligand Binding Ligands Machine Learning Malignant Neoplasms Measures Membrane Proteins Methods Modeling Molecular Molecular Conformation Molecular Profiling Mutation Neurologic Osteoporosis Outcome Pain Pharmaceutical Chemistry Pharmacology Pilot Projects Property Publications Publishing Reporting Research Resistance development Roentgen Rays Role Signal Transduction Site Site-Directed Mutagenesis Structure Synthesis Chemistry System Techniques Technology Therapeutic Tissues Validation Work X-Ray Crystallography addiction alternative treatment base cannabinoid receptor cheminformatics computational chemistry deep learning design drug development drug discovery endogenous cannabinoid system improved in silico in vivo innovation insight interest learning classifier neuroinflammation novel pharmacophore positive allosteric modulator pressure prevent prospective prototype receptor screening small molecule small molecule libraries spatiotemporal structural biology synthetic cannabinoid targeted treatment therapeutic target three dimensional structure tool virtual

项目摘要

Cannabinoid receptor subtype 2 (CB2) is a class-A family G protein-coupled receptor (GPCR), located primarily in immune-associated tissues but also in specific regions of the brain, and implicated in several inflammatory diseases and addiction. Drugs targeting CB2 are attractive treatment alternatives for chronic neurological pain and neuroinflammatory autoimmune diseases since they avoid deleterious psychotropic effects that are associated with CB1. While drug development efforts have been primarily focused on small molecules targeting the orthosteric site, limitations of poor selectivity, lack of efficacy, and development of resistance have hampered such effort. At present, there is great interests in identifying GPCR allosteric modulators that either enhance (positive allosteric modulators, or PAMs) or inhibit (negative allosteric modulators, or NAMs) agonist-induced receptor activity. PAMs/NAMs often exhibit improved subtype selectivity and spatiotemporal sensitivity, as well as potential biased signaling properties compared to orthosteric ligands. We have recently reported a 3.2 Å cryo-EM structure of the agonist-bound human CB2-Gi complex. Based on such progress, the overall goals of this proposal are to obtain a structural understanding of CB2 allosteric modulation and use our integrated computational and experimental medicinal chemistry/biology approaches to design and synthesize novel allosteric modulators for the development of CB2-specific small-molecules with potential to treat CB2-associated maladies. Thus, we first propose to elucidate the structural basis for the action of CB2 allosteric modulators by cryo-EM and X-ray crystallography approaches. To achieve the goal, we will advance our established methods for structural studies on CB2 to obtain structure of CB2 with known PAMs or NAMs. Subsequently, we plan to perform in silico design of novel CB2 allosteric modulators by our established molecular fingerprint machine-learning (ML) computing algorithms and receptor docking approaches, on basis of our reported chemogenomics cannabinoid molecular information database (CBID) and 3D CB2-Gi cryo-EM structure; a virtual allosteric modulator library will be constructed using our fragment-based design (FBD) method and our established ML-classifiers and features-ranking will be applied for selection of virtual hits. Results will be correlated with CB2 structure-based modulator design via adapting the structural information obtained from our recent CB2-Gi cryo-EM structure and our novel molecular complex characterizing system (MCCS) algorithm. Finally, we will carry out medicinal chemistry synthesis of CB2 PAM and NAM ligands and validate them by radiometric binding and cellular functional assays. With the proof-of-evidence of our recent discovery of a putative CB2 NAM, successful completion of these Aims will provide unprecedented structural information on CB2 allosteric pockets, identify promising new CB2 allosteric modulators, and help to elucidate CB2 signaling and pharmacology.

大麻素受体亚型 2 (CB2) 是 A 类 G 蛋白偶联受体 (GPCR)，主要位于存在于免疫区域相关组织中，也存在于特定的大脑中，并且与多种炎症有关针对 CB2 的药物是治疗慢性神经性疼痛的有吸引力的替代方案。和神经炎症性自身免疫性疾病，因为它们避免了有害的精神作用与 CB1 相关的药物开发工作主要集中在小分子靶向上。正位点、选择性差的局限性、缺乏功效和耐药性的发展阻碍了目前，人们对鉴定能够增强的 GPCR 变构调节剂非常感兴趣。（正变构调节剂，或 PAM）或抑制（负变构调节剂，或 NAM）激动剂诱导的 PAM/NAM 常常表现出改善的亚型选择性和时空敏感性以及时空敏感性。与正位配体相比，我们最近报道了 3.2 Å 冷冻电镜的潜在偏向信号特性。激动剂结合的人类 CB2-Gi 复合物的结构基于这些进展，该提案的总体目标。是为了获得对 CB2 变构调制的结构理解并使用我们的综合计算和实验药物化学/生物学方法来设计和合成新型变构调节剂开发具有治疗 CB2 相关疾病潜力的 CB2 特异性小分子。通过冷冻电镜和 X 射线晶体学阐明 CB2 变构调节剂作用的结构基础为了实现这一目标，我们将推进我们已建立的 CB2 结构研究方法，以获得随后，我们计划对新型 CB2 变构进行计算机设计。我们已建立的分子指纹机器学习 (ML) 计算算法和受体的调制器对接方法，基于我们报告的化学基因组学大麻素分子信息数据库（CBID）和 3D CB2-Gi 冷冻电镜结构；将使用我们的基于片段构建虚拟变构调制器库；设计 (FBD) 方法以及我们建立的 ML 分类器和特征排序将用于选择虚拟结果将通过调整结构信息与基于 CB2 结构的调制器设计相关联。从我们最近的 CB2-Gi 冷冻电镜结构和我们的新型分子复合物表征系统 (MCCS) 中获得最后，我们将进行CB2 PAM和NAM配体的药物化学合成并对其进行验证。通过放射结合和细胞功能测定，我们最近发现了一个假定的证据。 CB2 NAM，这些目标的成功完成将为 CB2 变构提供前所未有的结构信息口袋，识别有前途的新型 CB2 变构调节剂，并帮助阐明 CB2 信号传导和药理学。