The Comprehensive Center for Chemical Probe Discovery and Optimization at Scripps

MLPCN的目标是将高通量化学方法与最先进的遗传学(细胞、分子和体内生物学)相结合，通过多学科努力发现用于细胞和体内系统的概念验证(POC)分子探针。探测器有助于将生物医学的进步转变为对公共健康和生活质量的影响。斯克里普斯中心在MLSCN试验阶段的生产能力通过发布PubChem中的68种分析方法、优化5个分子靶点的POC探针以及同行评议发布的低纳摩尔效力、选择性和体内有效性的探针来定义新的生物和治疗靶点来证明。该中心保持了1536孔(&gt；20/年)的化验开发和uHTS实施的全部生产能力。&gt；80个数据集以每月2次完整文库分析的速度在PubChem中发表。仅在过去12个月中，就对8个不同目标类别的18个分子目标进行了27次uHTS活动(包括20个基于细胞的初级筛选)，使用发布的8种探测格式(来自13个外部PI)。内部发现项目被筛选在一个包含600,000个化合物的库中，超出了MLPCN的要求。这些数据是在油井体积(5-10ul)中以低成本获得的。该中心节省了NIH化合物的收集和控制成本，每年使用2ul的化合物(溶液)，用于24个初级筛查，只需要5ul的点选、再确认和10点滴定。形式包括离子通量和GPCR分析，报告基因，转录因子和核受体分析，酶分析，蛋白质-蛋白质和蛋白质-RNA相互作用和表型筛选。该中心目前每年通过重新合成、化学信息学、药物化学、二次和特异性分析实施以及筛选药代动力学/代谢研究来支持8个项目的点击到探针的化学合成，以实现有效的探针优化。该中心开发了跨基因家族和酶的高通量选择性图谱的新技术，以优化靶标选择性探针。该综合中心通过满足3(目标)支持大多数目标类的POC探头的生产发现。1：制定每年25项HTS就绪性测试。2：每年在300-50万个化合物中实施25个uHTS筛查，并向PubChem发布有质量保证的数据。3：优化对细胞和/或体内探针或POC的筛选命中，每年10-15个程序。该中心提供公共数据、工具和资源，以加强NIH路线图的成功和对人类健康的影响。

The goal of the MLPCN is to integrate high-throughput chemical approaches with state-of-the art genetics, (cellular, molecular and in vivo biology in a multi-disciplinary effort to discover of proof-of-concept (POC) molecular probes for cell and in vivo systems. Probes help transform biomedical advances to impact on public health and quality of life. Production capacity at The Scripps Center in the pilot MLSCN phase was demonstrated by publishing >68 assays in PUBCHEM, optimizing POC probes for 5 molecular targets and peer-reviewed publication of probes with low nanomolar potency, selectivity, and in vivo efficacy, to define novel biological and therapeutic targets. The Center sustained full production capabilities for assay development and for uHTS implementation in 1536-well format (>20/year). >80 data sets were published in PUBCHEM at a rate of 2 full library assays/month. In the past 12 months alone, 27 uHTS campaigns including 20 primary cell-based screens) for 18 molecular targets in 8 different target classes, using 8 detection formats (and from 13 external PIs) published. Internal discovery projects were screened on a single compound library of >600,000, exceeding MLPCN requirements. These data were achieved in well volumes (of 5-10 ul at low cost. The Center spares NIH compound collection and controls cost using 2 ul of compound (solution per year for 24 primary screens, and requiring only 5 ul for hit-picks, reconfirmation and 10 point titrations. Formats include ion flux and GPCR assays, reporter gene, transcription factor and nuclear receptor assays, enzyme assays, protein-protein and protein-RNA interactions and phenotypic screens. The Center currently supports hit-to-probe chemical synthesis across 8 projects/year by resynthesis, chemi-informatics, medicinal chemistry, secondary and specificity assay implementation and screening pharmacokinetics/ metabolism studies for efficient probe optimization. The Center developed novel technologies for high throughput selectivity profiling across gene families and enzymes for optimizing target-selective probes. This Comprehensive Center supports production discovery of POC probes for most target classes by meeting 3 (Aims. 1: Develop 25 assays/year to HTS readiness. 2: Implement 25 uHTS screens/year at 300-500,000 individual compounds and publish quality-assured data to PUBCHEM. 3: Optimize screening hits to probes or POC in cells and/or in vivo for 10-15 programs/year. The Center provides public data, tools and resources hat enhance the success of NIH Roadmap and impact on human health.