Scripps Research Institute Molecular Screen Center(RMI)

描述(申请人提供)：MLSCN建议的关键假设是，高通量化学方法与最先进的后基因组序列、细胞、分子和体内生物学相结合，为通过发现概念验证(POC)分子促进生物医学科学的进步提供了一种快速而简便的机制。此外，它的POC分子的发现将显著加快生物医学进步的应用，以提高公众健康和生活质量。在过去的两年里，TSRI和其他地方的实验努力提供了大量的同行评议论文，支持这些方法以极快的速度推进领域的能力。此外，它们还展示了良好集成过程的能力，以发现具有精致选择性、有效性和体内有效性的化学探针分子，并定义新的生物靶标。当现代高通量化学生物学可以与遗传方法相结合时，如目标基因的同源重组删除，或具有显著体外活性的化合物的表达谱，就会出现令人信服的结果。这些方法在调节淋巴细胞再循环和免疫抑制以及神经发生方面确定了生物控制的新关键点。TSRI中的分析专业知识涵盖了从全细胞钙通量分析、报告基因分析、酶分析、蛋白质-蛋白质相互作用和细胞分化读数在内的许多不同形式。我们进一步期待这一方法与更高吞吐量的分子生物学方法(如siRNA)协同作用，从而成为后基因组序列生物医学科学中科学动力的根本驱动力。因此，TSRI提案的主题是灵活、综合地发现跨目标类别的概念验证分子，重点是GPCR和细胞分化的途径。我们建议通过专门致力于这三个具体目标来做到这一点：目标1：建立一个分析实施小组，有能力在第二年和第三年每年优化20项分析。目标2：实施HTS，以筛选20项分析，每年每项分析至少100,000种化合物，并及时将有质量保证的数据传输到PubChem数据库。目标#3：发展药代动力学和探索性合成化学基础设施，以识别能够在基于细胞的系统或体内进行生物学概念验证(POC)的HITS或HIT衍生分子。执行这些目标将提供社区数据、工具和资源，以便将化学生物学工具传播到更广泛的生物医学研究领域，从而加强美国国立卫生研究院人类健康可测量终点路线图的成功和影响。

DESCRIPTION (provided by applicant): The key hypothesis of the MLSCN proposal is that high throughput chemical approaches, when integrated with state-of-the art post-genome sequence, cell, molecular and in vivo biology, provides a rapid and facile mechanism for enhancing the progress of biomedical science by the discovery of proof-of-concept (POC) molecules. Moreover, it POC molecule discovery will significantly speed up the application of biomedical advances to enhancing the public health and quality of life. Experimental efforts at TSRI and elsewhere have provided substantial peer-reviewed papers over the past 2 years that support the ability of these approaches to move fields forward with great rapidity. In addition, they exemplify the ability of a well integrated process to discover chemical probe molecules of exquisite selectivity, potency and in vivo efficacy, and to define novel biological targets. Compelling results emerge when modern high throughput chemical biology can be integrated with genetic approaches such as homologous recombinant deletion of target genes, or expression profiling of compounds with significant in vitro activity. These approaches have defined new key points of biological control in the regulation of lymphocyte recirculation and immunosuppression, as well as in neurogenesis. Assay expertise within TSRI has covered many different formats from whole cell calcium flux assays, reporter gene assays, enzyme assays, protein-protein interactions and cellular differentiation readouts. We further expect this approach to synergize with higher throughput molecular biological approaches such as siRNA, and therefore be a fundamental driver for scientific momentum in post-genome sequence biomedical science. The theme of the TSRI proposal is therefore the flexible, integrated discovery of proof-of-concept molecules across target classes, with an emphasis on GPCRs and pathways of cellular differentiation. We propose to do so by committing specifically to these three Specific Aims: Aim #1: Establish an Assay Implementation Group with the ability to optimize 20 assays per year during Years 2 and 3. Aim #2: Implement HTS to screen 20 assays and a minimum of 100,000 compounds per assay per year with transfer of the quality-assured data in a timely matter to PubChem database. Aim #3: development of pharmacokinetics and exploratory synthetic chemistry infrastructure to identify hits or hit-derived molecules capable of allowing biological proof of concept (POC) in cell-based systems or in vivo. Execution on these goals will provide community data, tools and resources for the dissemination of the tools of chemical biology to a broader segment of the biomedical research, thus enhancing the success and impact of the NIH Roadmap of measurable endpoints in human health.