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

• 批准号: 8418128
• 负责人: David C Swinney
• 金额: $ 27.69万
• 依托单位: INSTITUTE FOR RARE/NEGLECTED DISEASES
• 依托单位国家: 美国
• 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/8418128
• 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

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）：该项目的总体目标是提供 HTS 筛选测定和范例，以鉴定导致人类非洲锥虫病 (HAT) 或昏睡病的布氏锥虫的蛋白激酶抑制剂。 HAT 在撒哈拉以南非洲地区流行，每年夺去约 3 万人的生命，使约 6000 万人面临感染风险。 HAT 不是制药行业的优先事项，因此 NIH 和 WHO 将其归类为被忽视的热带病 (NTD)。迫切需要新的、有效的化疗。为了确定新的化疗方案，Kojo Wilmot-Mensa 博士 (UGA) 使用了一种化学蛋白质组学策略，使用激酶抑制剂拉帕替尼来识别寄生虫所必需的激酶。他确定了三种具有额外遗传验证的激酶。在初步工作中，这些激酶被表达、纯化并建立了证明活性的酶测定。为了发现 HAT 新药的目标，已经成立了一个多学科团队来开发这些激酶的 HTS 测定和筛选范例。该团队将采用一种策略，利用表型和分子方法进行药物发现，从而通过以下方式提高成功的可能性：1) 使用三种布氏锥虫激酶，2) 采用检测形式的全长蛋白质和底物，捕获 3) 不同的分子作用机制，例如缓慢解离、ATP 非竞争性抑制和阻断激酶激活。识别出的分子将被聚集，并评估代表在表型寄生测定中的活性。我们的多学科方法融合了寄生虫生物学和化学蛋白质组学（Kojo Wilmot-Mensa 博士，UGA）、药物发现和激酶酶学（David Swinney 博士，非营利性罕见和被忽视疾病药物发现研究所）、激酶 HTS 检测开发和药物发现（Chakk Ramesha 博士，Medhus Bio）以及蛋白质表达和生产（David 博士）的互补技能。 Chereau，96 种蛋白质）。在三年资助期结束时，我们期望提供经过验证的 HTS 测定和筛选范例，以提供具有成为药物的质量的化合物。我们预计通过该计划开发的检测方法将提交给 HTS 设施，并且我们希望使用 HTS 的结果作为向适当实体寻求额外资金以进行先导化合物开发的基础。