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，以多学科的努力发现细胞和体内系统的概念验证（POC）分子探针。探针有助于将生物医学进步转化为对公共健康和生活质量的影响。斯克里普斯中心在中试MLSCN阶段的生产能力通过在PUBCHEM上发表>68个测定、针对5个分子靶点优化POC探针以及同行评审的低纳摩尔效力、选择性和体内功效探针的出版物来证明，以定义新的生物和治疗靶点。该中心维持了全面的生产能力，用于检测开发和1536孔格式的uHTS实施（>20/年）。在PUBCHEM上以每月2次完整文库测定的速率发布了>80个数据集。仅在过去的12个月里，就有27个uHTS活动，包括20个主要的基于细胞的筛选），使用8种检测格式（来自13个外部PI），针对8个不同目标类别中的18个分子靶标。内部发现项目在超过600，000个的单一化合物库上进行筛选，超过了MLPCN的要求。这些数据是以低成本在孔体积（5-10 μ l）中获得的。该中心节省了NIH化合物收集和控制成本，每年使用2 ul化合物溶液进行24次初级筛选，仅需5 ul用于命中选择、再确认和10点滴定。形式包括离子通量和GPCR测定、报告基因、转录因子和核受体测定、酶测定、蛋白质-蛋白质和蛋白质-RNA相互作用以及表型筛选。该中心目前通过再合成、化学信息学、药物化学、次级和特异性分析实施以及筛选药代动力学/代谢研究支持8个项目/年的命中探针化学合成，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分析的新技术，用于优化靶选择性探针。该综合中心通过满足3个目标，支持大多数目标类别的POC探针的生产发现。1：每年开发25个检测试剂盒，以准备HTS。2：每年对300- 500，000种化合物进行25次uHTS筛选，并向PUBCHEM发布质量保证数据。3：在细胞和/或体内优化对探针或POC的筛选命中，持续10-15个程序/年。该中心提供公共数据，工具和资源，以提高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.