Cannabinoid CB2 Receptor Structure and Allosteric Modulators

大麻素 CB2 受体结构和变构调节剂

• 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
• 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

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 不仅存在于免疫相关组织中，而且存在于大脑的特定区域，并且与多种炎症有关 疾病和成瘾。靶向 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。虽然药物开发工作主要集中在小分子靶向 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 这样的努力。目前，人们对鉴定 GPCR 变构调节剂非常感兴趣，这些调节剂可以增强 （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 受体活性。 PAM/NAM 通常表现出改进的亚型选择性和时空敏感性，以及 与正构配体相比，潜在的偏向信号特性。我们最近报道了 3.2 Å 冷冻电镜 激动剂结合的人 CB2-Gi 复合物的结构。在此基础上，本提案的总体目标 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。因此，我们首先提出 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 接近。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 CB2 与已知 PAM 或 NAM 的结构。随后，我们计划进行新型 CB2 变构的计算机设计 我们建立的分子指纹机器学习 (ML) 计算算法和受体调制器 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；将使用我们基于片段的技术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 命中。通过调整结构信息，结果将与基于 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 算法。最后，我们将进行CB2 PAM和NAM配体的药物化学合成并进行验证 通过放射结合和细胞功能测定。随着我们最近发现的推定证据的证据 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。