In Situ Synthesis Of GPCR Ligands

GPCR 配体的原位合成

• 批准号: 8620213
• 负责人: Vadim Cherezov
• 金额: $ 20.61万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8620213
• 关键词: Adverse effects; Affinity; Antineoplastic Agents; Basal cell carcinoma; Behavior; Binding; Binding Sites; Biological; CXCR4 Receptors; Chemistry; Clinical Trials; Complex; Crystallization; Crystallography; Detergents; Development; Drug Targeting; Family; Funding; Future; G Protein-Coupled Receptor Genes; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Goals; Human; Human Genome; In Situ; Joints; Knowledge; Ligand Binding; Ligands; Lipids; Malignant Neoplasms; Malignant neoplasm of pancreas; Membrane; Membrane Proteins; Metastatic Prostate Cancer; Methods; Molecular Conformation; Myeloid Leukemia; Non-Small-Cell Lung Carcinoma; Opioid Receptor; Orphan; Pathway interactions; Pharmaceutical Preparations; Proteins; Protocols documentation; Route; Signal Transduction; Solid Neoplasm; Structure; System; Technology; Therapeutic; Time; Transducers; Work; anti-cancer therapeutic; antitumor agent; antitumor drug; base; chemical reaction; design; drug discovery; drug market; flexibility; human SMO protein; innovation; member; novel; novel therapeutics; pharmacophore; public health relevance; receptor; serotonin 5 receptor; small molecule; smoothened signaling pathway; therapeutic target; three dimensional structure; tool; transmission process

DESCRIPTION (provided by applicant): G-protein coupled receptors (GPCRs) constitute the largest family of membrane proteins in the human genome with approximately 800 members. GPCR signaling through multiple effector pathways has profound therapeutic implications, making these receptors the targets of ~40% of currently marketed drugs. Modulation of GPCR activity by small molecule ligands is emerging as a new strategy for development of anticancer therapeutics. However, design of new drugs with higher efficacy and lower side effects, and detailed understanding of GPCR mechanism of action are hampered by the limited structural information. Structural studies of many GPCRs are in turn limited by a scarcity of high-affinity ligands that stabilize GPCRs and enable their crystallization. The project attempts to develop a new platform using the in situ click chemistry approach to discover novel ligands that bind GPCRs to serve as tool compounds for structural and functional studies or as starting point for drug discovery. Target-guided synthesis approaches including in situ click chemistry, by which reactive fragments are joined in the binding sites of a biological target, have shown great promise in drug discovery but have not been applied to GPCRs. The challenges lie in the GPCRs' membrane disposition, their high conformational dynamics, low stability and low expression levels. The proposed platform is being developed targeting all GPCRs, with initial focus primarily on those known to be cancer targets. In the two years of this study, we will focus on smoothened (SMO) receptor which is an orphan receptor and a key signal transducer in the Hedgehog (Hh) signaling pathway activated during development, and thus is a target of a number of antitumor drugs in clinical trials. Our goal will be attained by achieving the following two aims: (1) Establish an in situ click chemistry platform to generate high-affinity GPCR ligands, (2) Characterize the interaction between selected ligands from Aim 1 and SMO by crystallography. The study will be supported by a team that had developed the GPCR Structure Determination Pipeline as well as its underlying technologies which have led to the structure determination of 12 GPCR structures since 2007.

描述（由申请人提供）：G蛋白偶联受体（GPCR）构成人类基因组中最大的膜蛋白家族，约有800名成员。通过多种效应子途径的GPCR信号传导具有深远的治疗意义，使这些受体成为当前销售药物的靶标。小分子配体对GPCR活性的调节正在成为抗癌疗法发展的新策略。但是，有限的结构信息阻碍了具有较高功效和副作用较低的新药的设计，以及对GPCR作用机制的详细理解。许多GPCR的结构研究又受到稳定GPCR并能够结晶的高亲和力配体的稀缺性限制。该项目试图使用原位点击化学方法来开发一个新的平台，以发现结合GPCR的新型配体，以作为结构和功能研究的工具化合物或作为药物发现的起点。目标引导的合成方法在内，包括原位点击化学，通过该方法将反应性片段连接在生物靶标的结合位点中，在药物发现中显示出很大的希望，但尚未应用于GPCR。挑战在于GPCR的膜处置，其高构象动力学，低稳定性和低表达水平。所提出的平台正在针对所有GPCR，最初的重点主要集中在已知的癌症靶标。在这项研究的两年中，我们将重点关注平滑的（SMO）受体，该受体是刺猬（HH）信号传导途径在发育过程中激活的怪异受体和关键信号传感器，因此是临床试验中多种抗肿瘤药物的目标。我们的目标将通过实现以下两个目的来实现：（1）建立一个原位点击化学平台以生成高亲和力的GPCR配体，（2）表征来自AIM 1中选定的配体与Crementallogrich的SMO之间的相互作用。这项研究将得到开发GPCR结构测定管道及其基础技术的团队的支持，该技术已导致自2007年以来的12个GPCR结构的结构确定。