Development of HTS assay and screening paradigm to discover new kinase inhibitors

开发 HTS 测定和筛选范例以发现新的激酶抑制剂

基本信息

批准号：
8652432
负责人：
KOJO A. MENSA-WILMOT
金额：
$ 30.67万
依托单位：
INSTITUTE FOR RARE/NEGLECTED DISEASES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8652432
关键词：
Adverse effects Africa South of the Sahara African Trypanosomiasis Basic Science Binding Sites Biochemical Biological Assay Biology Bite Blood - brain barrier anatomy Central Nervous System Diseases Chemical Structure Chemicals Clinical Development Disease Dissociation Dose Drug Industry Drug Targeting Eflornithine Enzymatic Biochemistry Enzymes Funding Gefitinib Genetic Goals Human Imatinib Infection Institutes Lead Left Length Malignant Neoplasms Marketing Medicine Melarsoprol Methodology Mission Molecular Molecular Conformation Molecular Mechanisms of Action Parasites Pentamidine Pharmaceutical Preparations Phosphotransferases Probability Production Property Protein Kinase Inhibitors Proteins Proteomics Risk Staging Suramin Therapeutic Therapeutic Index Translating Tropical Disease Trypanosoma brucei brucei Trypanosoma brucei gambiense Tsetse Flies Tumor stage United States National Institutes of Health Validation Work assay development base chemotherapy combat drug discovery enzyme substrate high throughput screening inhibitor/antagonist interdisciplinary approach kinase inhibitor lapatinib multidisciplinary mutant neglect novel protein expression protein kinase inhibitor public health relevance screening skills small molecule libraries success tool

项目摘要

DESCRIPTION (provided by applicant): The overall objective of the project is to provide a HTS screening assay and paradigm to identify protein kinase inhibitors for Trypanosoma brucei that causes Human African trypanosomiasis (HAT), or sleeping sickness. HAT is endemic in sub-Saharan Africa, claiming the lives of about 30000 people every year and putting approximately 60 million people at risk of infection. HAT is not a priority for the pharmaceutical industry and thus the NIH and WHO categorized it as a neglected tropical disease (NTD). The need for new, effective chemotherapy is urgent. In order to identify new chemotherapy Dr Kojo Wilmot-Mensa (UGA) has used a chemical proteomic strategy using the kinase inhibitor, lapatinib, to identify kinases essential to the parasite. He identified three kinases that have additional genetic validation. In preliminary work these kinases were expressed, purified and enzyme assays established that demonstrated activity. Toward the goal of discovering new medicines for HAT, a multidisciplinary team has been established to develop an HTS assay and screening paradigm for these kinases. The team will employ a strategy to take advantage of both phenotypic and molecular approaches to drug discovery and therefore increase the probability of success through the 1) use of three T. brucei kinases, 2) full-length protein and substrates in assay formats that will capture 3) different molecular mechanisms of action, such as slow-dissociation, ATP noncompetitive inhibition and blocking kinase activation. The molecules identified will be clustered and representatives evaluated for activity in phenotypic parasitic assays. Our multidisciplinary approach merges the complementary skills of parasite biology and chemical proteomics (Dr Kojo Wilmot-Mensa, UGA), drug discovery and kinase enzymology (Dr David Swinney, non-profit Institute for Rare and Neglected Diseases Drug Discovery), kinase HTS assay development and drug discovery (Dr Chakk Ramesha, Medhus Bio) and protein expression and production (Dr David Chereau, 96 Proteins). At the end of the three-year funding period we expect to deliver a validated HTS assays and screening paradigm to deliver compounds with the quality to become medicines. We anticipated that assays developed through this initiative will be submitted to HTS facilities and we expect to use the results from HTS as the basis for seeking additional funding from appropriate entities for lead compound development.

描述（由申请人提供）：该项目的总体目的是提供HTS筛查测定法和范式，以鉴定brucei锥虫瘤的蛋白激酶抑制剂，从而导致人类非洲人类非洲锥虫病（HAT）或睡眠疾病。帽子在撒哈拉以南非洲是地方性的，每年约有30000人的生命，并使约6000万人处于感染风险中。帽子不是制药行业的优先事项，因此是NIH，并将其归类为被忽视的热带疾病（NTD）。需要新的，有效的化学疗法的需要。为了鉴定新的化学疗法，Kojo Wilmot-Mensa博士（UGA）使用了使用激酶抑制剂拉帕替尼使用化学蛋白质组学策略来识别寄生虫必不可少的激酶。他确定了三个具有额外遗传验证的激酶。在初步工作中，这些激酶被表达，纯化和酶测定，并确定了活性。为了找到针对HAT的新药物的目标，已经建立了一个多学科团队，以开发这些激酶的HTS分析和筛选范式。 The team will employ a strategy to take advantage of both phenotypic and molecular approaches to drug discovery and therefore increase the probability of success through the 1) use of three T. brucei kinases, 2) full-length protein and substrates in assay formats that will capture 3) different molecular mechanisms of action, such as slow-dissociation, ATP noncompetitive inhibition and blocking kinase activation.确定的分子将被聚集，并评估代表在表型寄生分析中的活性。 Our multidisciplinary approach merges the complementary skills of parasite biology and chemical proteomics (Dr Kojo Wilmot-Mensa, UGA), drug discovery and kinase enzymology (Dr David Swinney, non-profit Institute for Rare and Neglected Diseases Drug Discovery), kinase HTS assay development and drug discovery (Dr Chakk Ramesha, Medhus Bio) and protein expression and production (Dr David Chereau，96蛋白）。在三年资金期结束时，我们希望提供经过验证的HTS测定法和筛选范式，以提供具有质量的化合物以成为药物。我们预计，通过该计划开发的分析将提交HTS设施，我们希望将HTS的结果作为寻求铅复合开发的适当实体的额外资金的基础。