Target-based Assays and Screening Strategies for Chemical Probe and Therapeutic Lead Discovery

该项目包括基于与疾病有关的特定蛋白质或核酸靶点的生物化学和靶点集中测定的开发。在分析和进展策略的背景下考虑测定设计，以使用高通量筛选技术评价各种化合物类别。有一个强烈的重点是方法开发研究，以提高检测和铅发现效率。作为对这些活动的补充，我们还探索和设计了询问复杂化学库的方法（例如，天然产物提取物，mRNA展示）。该计划的工作用于支持一系列赠款申请和原型项目。 以下是正在进行的： Radixin介导的cAMP效应子组装。cAMP是一种通用的第二信使，在每种细胞类型中被几种激素使用。它涉及正常生理和疾病的多个方面，因此是药物发现的有效但具有挑战性的目标。与D合作。Altschliche（U. Pittsburgh），我们正在研究特异性靶向cAMP依赖性细胞增殖中涉及的cAMP效应子-支架复合物的测定策略。 用小分子靶向蛋白质棕榈酰化。与A合作。Banerjee（NICHD，NIH），我们正在开发1536孔兼容的蛋白质棕榈酰酰基转移酶测定法，以评估潜在的酶抑制剂的化学库，作为可能的治疗线索，用于与这类酶相关的大量疾病。 这些化合物也被认为具有作为结构、功能和药理学探针的价值。 发现RNA小分子配体的策略。与N教授合作。Baird（U.我们正在探索检测设计，以探测小分子与非编码基因调控mRNA的相互作用。这项研究有可能发现新的抗生素或抗癌药物。 能够发现受体鸟苷酸环化酶Npr 1的新型小分子拮抗剂的测定开发。与M合作。Hoon（NIDCR，NIH），我们正在开发和测试新型的B型利钠肽（BNP）受体Npr 1检测方法。 最近的激动剂，脑钠肽被证明是需要的周围和脊髓神经之间的瘙痒感觉的传输。Npr 1检测将用于鉴定用于慢性瘙痒药物治疗的拮抗剂，慢性瘙痒是一种导致长期不懈的抓挠冲动的病症，显著降低患者的生活质量。 SIRPa-CD 47蛋白-蛋白相互作用。与T.米勒（范式转换治疗学）和D. Roberts（NCI，NIH）我们的目标是利用CD 47作为许多肿瘤中的分子靶点的广泛潜力，以创造保护正常组织免受化疗和放疗的治疗方法，同时差异化地增强这些治疗对肿瘤的影响。 我们目前正在重新设计一个中等通量的生化CD 47配体结合试验，以更高的通量系统能够筛选大量的分子。

This project includes the development of biochemical and target-focused assays based on specific protein or nucleic acid targets implicated in disease. The assay designs are considered in the context of analysis and progression strategies for evaluation of a wide range of compound classes using high throughput screening technologies. There is a strong emphasis on methods development research to advance assay and lead discovery efficiency. Complementing these activities we also explore and devise approaches for the interrogation of complex chemical libraries (e.g., natural product extracts, mRNA display). The work from this program is used to support a range of grant applications and prototype projects. The following are on-going: Radixin-mediated assembly of cAMP effectors. cAMP is a universal second messenger used by several hormones in every cell type. It is involved in multiple aspects of normal physiology and disease and thus a valid though challenging target for drug discovery. In collaboration with D. Altschuler (U. Pittsburgh) we are investigating assay strategies to specifically target a cAMP effector-scaffold complex involved in cAMP-dependent cell proliferation. Targeting protein palmitoylation with small molecules. In collaboration with A. Banerjee (NICHD, NIH) we are developing 1536-well compatible protein palmitoyl acyl transferase assays to evaluate chemical libraries for potential inhibitors of the enzyme as possible therapeutic leads for the large number of diseases to which this class of enzyme have been linked. These compounds are also anticipated to have value as structural, functional and pharmacological probes. Strategies for the discovery of small molecule ligands of RNA. In collaboration with Prof. N. Baird (U. Sciences) we are exploring assay designs to probe the interaction of small molecules with non-coding gene-regulatory mRNAs. The research has the potential of the discovery of novel antibiotics or anticancer agents. Assay development to enable discovery of novel small molecule antagonists of the receptor guanylate cyclase Npr1. In collaboration with M. Hoon (NIDCR, NIH) we are developing and testing novel assays of the b-type natriuretic peptide (BNP) receptor, Npr1. Recently the agonist, BNP was shown to be required for the transmission of itch sensation between peripheral and spinal cord nerves. The Npr1 assays will be used to identify antagonists for use in pharmacological treatments of chronic itch, a condition that results in long-term unremitting urge to scratch that significantly degrades the quality of life for sufferers. SIRPa-CD47 Protein-protein interaction. In collaboration with T. Miller (Paradigm Shift Therapeutics) and D. Roberts (NCI, NIH) our goal is to leverage the broad potential of CD47 as a molecular target in a number of tumors to create therapeutics that protect normal tissue from chemo and radiation therapy while differentially enhancing the effects of these therapies on the tumor. We are currently reengineering a medium throughput biochemical CD47 ligand binding assay to a higher throughput system capable of screening a larger number of molecules.