High Throughput Screening of Peptide Pharmaceuticals

多肽药物的高通量筛选

基本信息

批准号：
7325917
负责人：
ALEX CHENCHIK
金额：
$ 25.23万
依托单位：
CELLECTA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7325917
关键词：
Address Agonist Amino Acids Animal Model Applications Grants Bacteriophage T7 Bacteriophages Binding Biological Biological Assay Biology Cell Line Cell Separation Cell Surface Receptors Cell model Cells Chemicals Clinic Clinical Clinical Trials Cloning Vectors Collaborations Collection Compatible Complex Custom DNA Development Disease Disease model Dissection Drug Delivery Systems Drug Receptors Epitopes Exons Facility Construction Funding Category Foundations Funding Genes Genome Genomics Goals Green Fluorescent Proteins Growth Factor Hormones Human Human Genome Project Interleukin-1 Laboratories Lead Libraries Ligands Modeling Molecular Mus Oligonucleotides Peptide Library Peptide Phage Display Library Peptides Performance Phage Display Pharmaceutical Preparations Pharmacologic Substance Phase Phase I Clinical Trials Process Proteins Protocols documentation Range Reagent Reporter Research Research Institute Research Personnel Resources Screening procedure Services Signal Pathway Signal Transduction Pathway Small Molecule Chemical Library Specificity Surface System TNF gene Technology Testing Therapeutic Therapeutic Intervention Toxic effect Translational Research Validation base cancer cell chemokine commercialization cost cytokine cytotoxic design dimer drug discovery experience extracellular high throughput screening high throughput technology human disease improved next generation novel novel therapeutics particle peptidomimetics programs receptor scale up small molecule small molecule libraries success synthetic peptide therapeutic target therapy development tool

项目摘要

DESCRIPTION (provided by applicant): Despite the recent completion of the human genome project, an ostensibly more difficult post-genomic challenge will be the functional annotation of all human genes and integration of this information into a rational drug discovery process. Unfortunately, this is at present challenging, primarily due to the limited choice of suitable toolsets to rapidly delineate potential drug targets and screen new drugs en masse. Phage display is a unique tool for the discovery of peptide ligands, design of peptidomimetics, and biological validation of specific targets. Despite these successes, application of phage display technology for High Throughput Screening (HTS) of biologically active peptides in a disease-relevant cell model has not been developed so far. To address these issues, we will develop a novel experimental platform for cell-based HTS of peptide drugs as an alternative to HTS of synthetic peptide or chemical small molecule libraries. The long-term goal of the current grant application is the development and commercialization of next- generation HTS technology for the simultaneous screening of millions of peptide compounds for biological activity with pooled phage display libraries, concomitantly with a wide range of reporter cell lines compatible with this HTS technology of peptide leads. The technologically simple and cost-effective approach of cell- based HTS with pooled phage libraries displaying millions of peptide compounds, which far exceeds the complexity of small molecule libraries, will change current drug discovery strategies and accelerate the development of novel therapeutics. We propose to use our novel bioactive peptide HTS resource to delineate the processes which underlie deregulated proliferation in cancer cells, and to apply this technology to identify cytotoxic peptides specific for cancer cells. Under Phase I funding, we initially propose to develop a pooled phage library displaying peptides for 500 pharmaceutically relevant cytokines, growth factors, chemokines, and hormones, as well as technology for HTS of biologically active phage displayed peptides in a cell-based assay. In collaboration with our biology consultants at the Cleveland Clinical Foundation, we will subsequently validate this library in transcriptional reporter cell line models, to identify effector peptides of NF-?B signaling pathway. The combination of phage display technology with cell-based HTS will transform phage display technology into a powerful tool for identification and validation of novel biologically relevant drugs. Moreover, we envisage a major impact on the molecular dissection of human disease mechanisms. For example, these reagents harbor considerable promise to identify new targets for therapeutic intervention, and the development of increasingly relevant paradigms for drug discovery. As a result, we foresee that these toolsets will significantly improve the efficiency, economy, and ease of performing HTS of lead peptide compounds, and will provide both academic and industrial researchers with preferred, cost-effective alternatives to existing small molecule libraries. The ultimate goal of the proposed project is to develop and make commercially available new, powerful drug discovery tools: a technology for high throughput screening of peptide pharmaceuticals in a cell-based assay with a set of genome-wide receptor ligand phage display libraries and transcriptional reporter cell lines. We propose to validate and apply these tools for discovery peptide agonists and antagonists of NFkB signaling pathway. The developed high throughput screening tools and technologies will significantly improve the efficiency of translational research related to molecular dissection of diverse human disease mechanisms, development of new pharmaceuticals, and therefore, have major implications for improving drug discovery research.

描述（由申请人提供）：尽管人类基因组项目最近完成，表面上更困难的基因组挑战将是所有人类基因的功能注释以及将此信息整合到合理的药物发现过程中。不幸的是，目前这具有挑战性，这主要是由于合适的工具集有限的选择，可以快速描绘潜在的药物靶标和筛查新药。噬菌体显示是发现肽配体，肽仪设计和特定靶标生物学验证的独特工具。尽管取得了这些成功，但迄今为止尚未开发出在与疾病相关的细胞模型中生物活性肽高吞吐量筛查（HTS）的应用。为了解决这些问题，我们将开发一个新型的实验平台，用于基于细胞的肽药物的HTS，以替代合成肽或化学小分子库的HTS。当前赠款应用程序的长期目标是下一代HTS技术的开发和商业化，用于同时筛选数百万肽化合物，用于生物活性，并与合并的噬菌体展示库一起筛选，同时筛选了与这种与这种与这种与这种HTS肽铅技术相兼容的广泛记者细胞系。基于噬菌体的噬菌体库的基于细胞的HT的技术简单且具有成本效益的方法，这些方法显示出数百万肽化合物（远远超过小分子库的复杂性）将改变当前的药物发现策略，并加速新型治疗剂的发展。我们建议使用我们的新型生物活性肽HTS资源来描述癌细胞中消除癌细胞增殖的过程，并应用该技术以鉴定针对癌细胞的细胞毒性肽。在第一阶段的资金下，我们最初提议开发一个汇集的噬菌体文库，该噬菌体图书馆显示500种药物相关的细胞因子，生长因子，趋化因子和激素，以及用于生物活性噬菌体HTS的技术，在基于细胞的分析中显示肽。与我们在克利夫兰临床基金会的生物学顾问合作，我们随后将在转录报告细胞系模型中验证该库，以识别NF-？B信号通路的效应肽。噬菌体展示技术与基于细胞的HTS的组合将把噬菌体显示技术转变为一种强大的工具，用于识别和验证新型生物学相关药物。此外，我们设想对人类疾病机制的分子解剖产生重大影响。例如，这些试剂具有巨大的承诺，可以识别治疗干预的新目标，并开发了越来越多的相关药物发现范式。结果，我们预见到这些工具集将显着提高效率，经济性和易于执行铅肽化合物的HTS，并将为学术和工业研究人员提供现有小分子库的优先且具有成本效益的替代方案。拟议项目的最终目标是开发和制造商业上可用的新型功能强大的药物发现工具：一种在基于细胞的测定中，具有一组全基因组受体配体显示库和转录记者细胞系中的基于细胞的肽药物的高吞吐量筛查技术。我们建议验证并将这些工具应用于NFKB信号通路的发现肽激动剂和拮抗剂。开发的高吞吐量筛查工具和技术将显着提高与分子解剖有关的人类疾病机制，新药的开发，因此对改善药物发现研究具有重要意义。