Multiplexed Measurement of Psychoactive Drug Selectivity Profiles

描述（由申请方提供）：本II期提案的主题是完成CNS相关受体的GPCR筛选组，从而能够发现作用于多个GPCR靶点的治疗候选物。大多数内源性GPCR配体激活多种受体。这些配体通过“选择性非选择性”实现生物学功效;由此受体的小子集被特定配体不同程度地激活。同样，许多治疗神经系统和物质滥用障碍的有效药物之所以有效，仅仅是因为它们调节多个靶点。不幸的是，目前的药物发现技术只允许一次发现一个受体，有效地留下任何有益的“脱靶”受体相互作用的机会。为了有目的地鉴定调节多种感兴趣受体的化合物，需要同时分析这些反应的系统。在第一阶段研究中，Primont开发了一种新的GPCR检测和细胞条形码系统，能够在微量滴定板的单个孔中同时筛选8个GPCR靶标。该II期提案进一步将神经系统疾病小组扩大到54个靶点，提供了数十种经验证的CNS药物靶点的复合作用的全球视图。将优化用于检测气相聚合酶链式反应激活的技术，以进行高通量筛选，并使数据分析自动化，以便能够在每个筛选日筛选4 000个威尔斯孔（> 100 000个数据点）。该系统将以1500种化合物的中试筛选为基准，以确定大规模平行GPCR筛选的可行性。将目标扩展到54，再加上本提案中概述的适当筛选指标的演示，将为将该技术商业化提供基础，作为有目的地发现神经GPCR目标的多效性配体的新型筛选平台。 公共卫生相关性：神经系统疾病涉及具有高度冗余功能的受体的复杂网络，使得使用传统的“一种药物，一种受体”模型难以实现治疗功效。本文所述的系统有助于在广泛的CNS相关靶标上进行高效的化合物筛选，以鉴定已知药物的意外活性，并能够发现同时靶向多种受体的药物。

DESCRIPTION (provided by applicant): The subject of this Phase II proposal is the completion of a GPCR screening panel for CNS- related receptors, enabling the discovery of therapeutic candidates acting at multiple GPCR targets. Most endogenous GPCR ligands activate multiple receptors. These ligands achieve biological efficacy through a "selective non-selectivity"; whereby a small subset of receptors is activated to varying degrees by a specific ligand. Similarly, many effective drugs for neurological and substance abuse disorders are effective only because they modulate multiple targets. Unfortunately, current drug discovery technologies only permit ligand discovery one receptor at a time, effectively leaving any beneficial "off-target" receptor interactions to chance. In order to purposefully identify compounds that modulate multiple receptors of interest, a system is needed that profiles these responses simultaneously. In Phase I studies, Primity developed a novel GPCR assay and cellular barcoding system that enabled the simultaneous screening of eight GPCR targets in a single well of a microtiter plate. This Phase II proposal further expands the Neurological Disease Panel to 54 targets providing a global view of compound action across dozens of validated CNS drug targets. The techniques used for detecting GPCR activation will be optimized for high-throughput screening, and the data analysis automated, to enable screening of 4,000 wells (>100,000 data points) per screening day. The system will be benchmarked with a pilot screen of 1500 compounds to establish feasibility of massively parallel GPCR screening. The extension of targets to 54, coupled with the demonstration of the appropriate screening metrics outlined in this proposal, will provide the foundation for commercializing this technology as a novel screening platform for the purposeful discovery of pleiotropic ligands for neurological GPCR targets. PUBLIC HEALTH RELEVANCE: Neurological diseases involve a complex web of receptors with highly redundant functions making it difficult to achieve therapeutic efficacy using a traditional 'one drug, one receptor' model. The system described here facilitates highly efficient compound screening across a broad range of CNS- related targets to identify unexpected activities of known drugs and enable the discovery of drugs that simultaneously target multiple receptors.