A Pharmacochaperone-based strategy for identifying chemical probes of brain-derived orphan GPCRs

项目总结 没有什么地方比治疗中更迫切地需要改变疾病的治疗化合物了 精神健康障碍。尽管在药物和行为干预方面取得了重大进展，但心理干预 精神分裂症、抑郁症和焦虑症等疾病与药物滥用障碍相结合仍然是一种 给社会带来沉重且日益沉重的负担。孤儿G蛋白偶联受体(OGPCRs)代表 这些疾病潜在的新药靶点尚未开发，但缺乏药理工具 需要验证这些靶点，以防止它们的治疗性利用。作为利用这一技术的第一步 为了开发新的药物类别，我们建议寻找与oGPCRs结合的化合物，并可以 被用来验证它们是否为药物靶标。利用最新的高分辨率基因表达图谱 重要的大脑回路，我们确定了27个oGPCRs，它们在人脑中的离散表达模式 让他们成为高价值的神经科学药物靶点。这些都是根据以下正文进一步确定优先顺序的 每个受体可获得的生物学数据(例如，遗传学、与疾病的联系)，目的是侧重于 与心理健康关系最大的GPCRs。对于前十个优先目标，我们将进行高吞吐量 使用一种创新的配体诱导正向运输分析来筛选包含约50,000种化合物的文库 我们最近发表了。在这个实验中，每个oGPCR都被设计成保留在内质网中 通过配体的结合实现前向传输的一种状态。因此，oGPCR的贩运是 使用分离的β-半乳糖苷酶报告系统进行监测。通过这种方式，结合和诱导的化合物(命中) OGPCR的构象变化将被识别出来。使用一组交叉和计数器屏幕分析， 命中将得到确认，并显示与本地oGPCR有关。将利用药物化学来改进 效力和选择性，并产生具有物理化学属性的化学探针 细胞分析探索靶向oGPCRs的功能和药理学。探头的选择性将是 使用不同的靶标进行评估，包括GPCRs、离子通道、核受体和酶。 在人类IPSC来源的神经元培养中，如果特定的oGPCR高表达，我们将研究 使用微电极阵列检测系统检测特定受体的探针影响神经网络的能力 我们开发了。我们预计，这些努力将产生针对高度优先的gpcr的10个新的探测。 我们的目的是迅速将这些信息分发给科学界，以加速对这些信息的理解 大脑丰富了oGPCRs，有助于确认它们是治疗心理障碍的药物靶点。

PROJECT SUMMARY Nowhere is the need for disease modifying therapeutic compounds more urgently needed than in the treatment of mental health disorders. Despite significant advances in pharmaceutical and behavior interventions, mental illnesses such as schizophrenia, depression and anxiety, combined with substance abuse disorders remain a significant and growing burden to society. Orphan G-protein coupled receptors (oGPCRs) represent an untapped reservoir of potential new drug targets for these diseases but the lack of pharmacological tools needed to validate these targets prevents their therapeutic exploitation. As a first step towards tapping this reservoir to develop new classes of medicines, we propose to find compounds that bind the oGPCRs and can be used to validate them as drug targets. Leveraging recent high-resolution gene expression maps of important brain circuits, we identified 27 oGPCRs whose discrete patterns of expression in the human brain make them high value neuroscience drug targets. These were further prioritized based on the body of biological data available for each receptor (e.g. genetics, linkage to disease) with the goal of focusing on oGPCRs of greatest relevance to mental health. For the top 10 priority targets we will conduct high-throughput screens against a library of ~50,000 compounds using an innovative ligand-induced forward trafficking assay we recently published. In this assay, each oGPCR is engineered to be retained in the endoplasmic reticulum in a state in which forward transport is enabled by the binding of a ligand. Trafficking of the oGPCR is then monitored using a split beta-galactosidase reporter system. In this way, compounds (hits) that bind and induce a conformational change in the oGPCR will be identified. Using a panel of cross and counter screen assays, hits will be confirmed and shown to engage native oGPCR. Medicinal chemistry will be used to improve potency and selectivity, and to generate chemical probes with physicochemical attributes suitable for use in cellular assays exploring the function and pharmacology of target oGPCRs. Selectivity of probes will be evaluated using a diverse panel of targets including GPCRs, ion channels, nuclear receptors and enzymes. Where a specific oGPCR is highly expressed in human iPSC-derived neuronal cultures, we will investigate the ability of probes to the specific receptor to affect neural networks using a microelectrode array assay system we developed. We anticipate that these efforts will generate >10 novel probes against high priority oGPCRs. Our intention is to rapidly distribute these to the scientific community to accelerate the understanding of these brain enriched oGPCRs, contributing to their validation as drug targets for psychological disorders.