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

项目概要 没有什么比在治疗中更迫切需要改变疾病的治疗化合物了 心理健康障碍。尽管在药物和行为干预方面取得了重大进展，但心理 精神分裂症、抑郁症和焦虑症等疾病以及药物滥用障碍仍然是一个常见问题 给社会带来重大且日益严重的负担。孤儿 G 蛋白偶联受体 (oGPCR) 代表 这些疾病的潜在新药物靶点尚未开发，但缺乏药理学工具 验证这些目标所需的信息可以防止它们的治疗利用。作为利用这一点的第一步 为了开发新类别的药物，我们建议寻找结合 oGPCR 的化合物，并且可以 用于验证它们作为药物靶点。利用最近的高分辨率基因表达图 重要的大脑回路，我们鉴定了 27 个 oGPCR，其在人脑中表达的离散模式 使它们成为高价值的神经科学药物靶标。这些根据主体进一步确定了优先顺序 每个受体的可用生物学数据（例如遗传学、与疾病的联系），目标是关注 oGPCR 与心理健康最相关。对于前10个优先目标我们将进行高通量 使用创新的配体诱导的正向运输测定对约 50,000 种化合物的库进行筛选 我们最近发布了。在此测定中，每个 oGPCR 都被设计为保留在内质网中 通过配体的结合实现正向运输的状态。 oGPCR 的贩运是 使用分裂β-半乳糖苷酶报告系统进行监测。通过这种方式，结合并诱导的化合物（命中） oGPCR 中的构象变化将被识别。使用一组交叉和反筛选分析， 命中将被确认并显示与本地 oGPCR 结合。药物化学将用于改善 效力和选择性，并生成具有适用于的理化属性的化学探针 探索目标 oGPCR 的功能和药理学的细胞测定。探针的选择性为 使用多种靶标进行评估，包括 GPCR、离子通道、核受体和酶。 当特定的 oGPCR 在人类 iPSC 衍生的神经元培养物中高度表达时，我们将研究 使用微电极阵列测定系统对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超过 10 种针对高优先级 oGPCR 的新型探针。 我们的目的是将这些内容快速分发给科学界，以加速对这些内容的理解 大脑中丰富的oGPCRs，有助于它们作为心理疾病药物靶点的验证。

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.