Molecular Targets Development Program

分子靶点开发计划

• 批准号: 8554068
• 负责人: james mcmahon
• 金额: $ 141.78万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8554068
• 关键词: ABCG2 gene; Algae; Animals; Antiviral Agents; Binding; Biochemical; Biological; Biological Assay; Biological Factors; Cells; Cessation of life; Chemicals; Clinical; Clinical Trials; Collaborations; Computing Methodologies; Cytotoxic agent; DNA; DNA Repair Enzymes; Data; Development; Generations; Genomics; Goals; HIV; Investigation; Laboratories; Lead; Libraries; Ligands; Liquid substance; MDM2 gene; Malignant Neoplasms; Marine Invertebrates; Microbe; Molecular; Molecular Biology; Molecular Target; Multi-Drug Resistance; Natural Products Chemistry; Online Systems; Outcome; Pathway interactions; Phenotype; Phosphorylated Peptide; Plants; Polo-Box Domain; Principal Investigator; Program Development; Protein Chemistry; Proteins; Proteomics; Relative (related person); Research; Research Infrastructure; Resources; Ribonuclease H; Sampling; Screening Result; Screening procedure; Source; Specificity; Synthesis Chemistry; System; TNFSF10 gene; Techniques; Transcription Factor AP-1; Tumor Suppressor Proteins; Validation; Work; analog; assay development; base; cancer therapy; cell killing; cellular targeting; design; follow-up; high throughput screening; human PLK1 protein; inhibitor/antagonist; interest; mucosa-associated lymphoid tissue; neoplastic cell; novel; pre-clinical; receptor; transcription factor; ubiquitin-protein ligase

Assay Development and Screening Section, James McMahon, Principal InvestigatorGoals: Assay development efforts focused on the configuration, optimization and validation of high-throughput assays to screen pure compound libraries and natural product extracts for modulators of specific molecular and cellular targets and associated cellular phenotypes. Preliminary characterization of screen hits for better prioritization in downstream follow-up. Accomplishments: Nine assays developed, 11 high-throughput screening (HTS) campaigns completed and 10 assays performed in support of natural products chemistry. Preliminary biological characterization work done on several hits from completed HTS campaigns. One lead compound obtained from HTS is currently in clinical trial while several others are under preclinical development.Protein Chemistry and Molecular Biology Section, Barry OKeefe, Principal Investigator Goals: Investigation of specific protein-ligand interactions to assess their applicability to potential screening approaches. Development of cell-free high throughput screens robust enough to tolerate crude natural product extracts and sufficiently predictive to guide the identification and purification of bioactive compounds. Post-screen biochemical characterization of active compounds to determine specificity, mechanism of inhibition and specific binding interactions. Isolation and characterization of antiviral proteins from natural product extracts. Accomplishments: Developed biochemical screens for inhibitors of the E3 ubiquitin ligase MDM2, tyrosyl-DNA phosphodiestease 1, the interaction of the polo-box domain of polo-like kinase 1 and its phosphorylated peptide ligand, and for the novel paracaspase mucosa-associated lymphoid tissue transformation protein 1. The PCMBS/OKeefe also biochemically evaluated hit compounds resulting from screens for inhibitors MDM2, the anti-HIV target RNase H and tyrosyl-DNA phosphodiestease 1. Finally, the PCMBS/OKeefe characterized the activity of several new classes of anti-HIV proteins from natural product extracts. Chemical Diversity Development Section, John Beutler, Principal Investigator Goals: Provide chemical diversity to MTL HTS screening assays by accessing multiple sources of pure compounds and selected natural product extracts. Isolation and characterization of novel anti-cancer natural products from cell-based assays and their preclinical development.Accomplishments: Initiated collaborations with synthetic chemistry groups and natural product groups to obtain screening samples, and established a library of 261,223 samples for screening, including extracts, prefractionated samples, pure natural products and pure synthetics. Identified hits from diverse external sources and obtained resupply of hit material and analogues in multiple screening projects including AP-1, Pdcd4, and Plk-1. Developed a web based assay and sample management system which allows MTL staff to manage sample libraries, design assay plates and collect and analyze HTS data. Interfaced sample management system directly with sample storage and liquid handling infrastructure. Preclinical development of both the englerins and schweinfurthins. Natural Products Chemistry Section, Kirk Gustafson, Principal Investigator Goals: Bioassay-directed isolation and structural elucidation of bioactive metabolites from extracts of marine invertebrate animals and algae, terrestrial plants, and microbes. Generation of an extract prefractionation library for more effective high-throughput screening outcomes. Adaptation and application of contemporary techniques for assigning the relative and absolute configuration of novel lead compounds. Provision of active natural products to support mechanism of action, target identification, and preclinical development studies.Accomplishments: Isolation and identification of natural products that inhibit a wide variety of molecular targets including transcription factors (AP-1 and HIF-2), tumor suppressors (Pdcd4, NF1), a multidrug resistance transporter (ABCG2), and a DNA repair enzyme (Tdp1). New compounds were discovered that synergistically enhance tumor cell killing by the TRAIL death receptor ligand. In addition, selective cytotoxic agents and anti-HIV natural products were identified using phenotypic screens. An extract prefractionation library with >100,000 samples was generated and shown to be a value-added screening resource. Structural and stereochemical assignments of diverse natural product chemotypes were accomplished using a combination of chemical manipulations, NMR analyses, CD studies, and computational methods.

测定开发和筛选部分，詹姆斯·麦克马洪（James McMahon），主要研究人员：分析开发工作重点是对筛选纯净化合物文库的构型，优化和验证，以筛选纯净化合物文库和天然产物提取物，用于特定分子和细胞靶标和相关细胞表型的调节剂。屏幕击中的初步表征可在下游随访中更好地优先级。成就：开发了9种测定，完成了11个高通量筛查（HTS）活动，并进行了10种测定法，以支持天然产品化学。初步的生物学特征工作对完成的HTS运动进行了几次打击。从HTS获得的一种铅化合物目前正在临床试验中，而另一种则处于临床前开发。蛋白质化学和分子生物学部分，Barry Okeefe，首席研究者目标：研究特定蛋白质 - 配体相互作用的研究，以评估其对潜在筛查方法的适用性。无细胞的高吞吐量筛选的发展足以耐受天然产物提取物，并充分预测，以指导生物活性化合物的鉴定和纯化。活性化合物的屏幕后生化表征，以确定特异性，抑制机制和特定的结合相互作用。从天然产物提取物中分离和表征抗病毒蛋白。 Accomplishments: Developed biochemical screens for inhibitors of the E3 ubiquitin ligase MDM2, tyrosyl-DNA phosphodiestease 1, the interaction of the polo-box domain of polo-like kinase 1 and its phosphorylated peptide ligand, and for the novel paracaspase mucosa-associated lymphoid tissue transformation protein 1. The PCMBS/OKeefe also由抑制剂MDM2，抗HIV靶RNase H和酪酶-DNA磷酸二次蛋白酶1的生化评估的HIT化合物1。最后，PCMBS/Okeefe表征了来自天然产物提取物的几种新型抗HIV蛋白的活性。化学多样性开发部分，约翰·贝特勒（John Beutler），主要研究者目标：通过访问多种纯化合物和选定的天然产品提取物的多种来源，为MTL HTS筛选分析提供化学多样性。从基于细胞的测定中及其临床前开发的新型抗癌天然产物的隔离和表征。从不同的外部来源中识别出命中，并在包括AP-1，PDCD4和PLK-1在内的多个筛选项目中获得了命中材料和类似物的补充。开发了一个基于Web的测定和样本管理系统，该系统允许MTL员工管理样本库，设计测定板并收集和分析HTS数据。直接将样品管理系统与样品存储和液体处理基础设施接口。 Englerins和Schweinfurthins的临床前发展。天然产品化学部分，柯克·古斯塔夫森（Kirk Gustafson），主要研究者的目标：从海洋无脊椎动物和藻类，陆生植物，陆生植物和微生物提取物中对生物测定的分离和结构阐明生物活性代谢物。生成提取物预分库文库，以进行更有效的高通量筛查结果。当代技术的适应和应用，用于分配新型铅化合物的相对和绝对构型。提供活跃的天然产品以支持作用机制，目标识别和临床前开发研究。章节：隔离和鉴定天然产物，这些天然产物抑制包括转录因子（AP-1和HIF-2），肿瘤抑制器（PDCD4，NF1），多种分子靶标（PDCD4，NF1），多种抗性抗性转运符（ABCG2）和ADNA（dna）和A dna（dna）。发现新的化合物是通过Trail Geath受体配体协同增强肿瘤细胞杀死的。此外，使用表型筛选鉴定出选择性的细胞毒性剂和抗HIV天然产物。生成了带有> 100,000个样品的提取预分库库，并显示为增值筛选资源。使用化学操作，NMR分析，CD研究和计算方法的组合完成了不同天然产物化学型的结构和立体化学分配。