Targeting trypanosomatid deoxyhypusine synthase

靶向锥虫脱氧马匹氨酸合酶

• 批准号: 9221920
• 负责人: Margaret A. Phillips
• 金额: $ 24.3万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9221920
• 关键词: Active Sites; African Trypanosomiasis; Amino Acids; Basic Amino Acids; Biochemical; Biological Assay; Biological Process; Brain; Cells; Chagas Disease; Chemicals; Disease; Drug Kinetics; Drug Targeting; Entamoeba; Enzymes; Eukaryota; Eukaryotic Cell; Eukaryotic Initiation Factors; Evaluation; Genetic Translation; Goals; Growth; Human; Human Cell Line; Impairment; In Vitro; Infection; Lead; Leishmania; Leishmaniasis; Libraries; Link; Lysine; Mass Spectrum Analysis; Metabolism; Modeling; Modification; Mus; Parasites; Pathogenicity; Penetration; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacotherapy; Phase; Polyamines; Property; Series; Solubility; Spermidine; Structure; Testing; Toxic effect; Trypanosoma brucei brucei; Trypanosoma cruzi; Validation; base; cell growth; cell killing; combat; cytotoxicity; deoxyhypusine synthase; gene product; high throughput screening; hypusine; in vitro Assay; in vivo; inhibitor/antagonist; killings; meetings; mouse model; neglected tropical diseases; novel; novel strategies; paralogous gene; pre-clinical; programs; resistance mechanism; small molecule; small molecule inhibitor; translation factor; treatment program; Active Sites; African Trypanosomiasis; Amino Acids; Basic Amino Acids; Biochemical; Biological Assay; Biological Process; Brain; Cells; Chagas Disease; Chemicals; Disease; Drug Kinetics; Drug Targeting; Entamoeba; Enzymes; Eukaryota; Eukaryotic Cell; Eukaryotic Initiation Factors; Evaluation; Genetic Translation; Goals; Growth; Human; Human Cell Line; Impairment; In Vitro; Infection; Lead; Leishmania; Leishmaniasis; Libraries; Link; Lysine; Mass Spectrum Analysis; Metabolism; Modeling; Modification; Mus; Parasites; Pathogenicity; Penetration; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacotherapy; Phase; Polyamines; Property; Series; Solubility; Spermidine; Structure; Testing; Toxic effect; Trypanosoma brucei brucei; Trypanosoma cruzi; Validation; base; cell growth; cell killing; combat; cytotoxicity; deoxyhypusine synthase; gene product; high throughput screening; hypusine; in vitro Assay; in vivo; inhibitor/antagonist; killings; meetings; mouse model; neglected tropical diseases; novel; novel strategies; paralogous gene; pre-clinical; programs; resistance mechanism; small molecule; small molecule inhibitor; translation factor; treatment program

Project Description. Trypanosomatid parasites are the causative agents of human African trypanosomiasis (HAT), Leishmaniasis and Chagas disease, all of which are listed by the WHO as Neglected tropical diseases (NTDs). Collectively 18-20 million people are infected with one of these parasites, yet drug therapies remain inadequate for all three diseases. Discovery of a drug-target that could be exploited against all three parasites, while not showing toxicity against the host, would provide a robust new approach to combat these diseases. Eukaryotes universally require a translation factor called eIF5A for cell growth. To be active eIF5A must be post-translationally modified on a conserved lysine reside to generate a novel amino acid called hypusine. The hypusine modification of eIF5A is catalyzed by deoxyhypusine synthase (DHS) using spermidine as a substrate. DHS is an essential enzyme in all eukaryotes including the trypanosomatids. We previously demonstrated that DHS from trypanosomatids and Entamoeba have uniquely evolved to require heterotetramer formation between two paralogous gene products to generate the active enzyme. This unusual configuration of these parasite DHSs suggests that it may be feasible to identify selective inhibitors of the parasite enzymes that don't inhibit human DHS. The goal of this proposal is to determine if drug-like inhibitors of DHS that are selective for the trypanosomatid enzymes can be identified and if so to advance them during a lead optimization program for the treatment of HAT. In the R21 phase of this proposal we plan to develop a high throughput screen (HTS) compatible assay for T. brucei DHS and to use this assay to screen the UT Southwestern 200,000 compound library for T. brucei DHS inhibitors. We will perform hit validation on identified inhibitors and determine if the identified inhibitors show good selectivity versus human DHS. If trypanosomatid specific DHS inhibitors with good drug like properties can be identified than in the R33 phase of the proposal we plan a hit to lead optimization of at least one series with appropriate properties to be advanced for the treatment of HAT. Identified DHS inhibitors will also be tested at various stages during hit validation and lead optimization against T. cruzi and Leishmania to determine if they have the potential to be developed as a broad-spectrum treatment for all three trypanosomatid-based diseases.

项目描述。 锥虫寄生虫是人类非洲锥虫病（HAT）的病原体， 利什曼病和查加斯病，所有这些都被世卫组织列为被忽视的热带疾病 （NTD）。总共有1,8-2000万人感染了其中一种寄生虫，但药物疗法感染 对所有三种疾病保持不足。发现可以利用所有人的药物目标 三个寄生虫虽然不对宿主表现出毒性，但将为新方法提供强大的新方法 对抗这些疾病。真核生物普遍需要一个称为EIF5A的翻译因子来用于细胞生长。 要活跃的eif5a必须在保守的赖氨酸上进行翻译后修改以生成 新型氨基酸称为氢酸。 deoxyhypusine催化EIF5A的无限修饰 使用精子作为底物的合成酶（DHS）。 DHS是所有真核生物中必不可少的酶 锥虫。我们以前证明了来自锥虫和Entamoeba的DHS 独特进化以在两个寄生基因产物之间形成异驱精剂才能产生 活性酶。这些寄生虫DHSS的这种不寻常的配置表明，它可能是可行的 鉴定不抑制人DHS的寄生虫酶的选择性抑制剂。目标的目标 建议是确定对锥虫酶有选择性的DHS的药物样抑制剂 可以识别，如果可以在铅优化计划中以促进帽子的治疗。 在此提案的R21阶段，我们计划开发高吞吐量屏幕（HTS）兼容测定 对于T. Brucei DHS，并使用此测定法筛选了UT西南200,000库。 Brucei DHS抑制剂。我们将对已识别的抑制剂进行命中验证，并确定是否是否 鉴定的抑制剂与人类DHS表现出良好的选择性。如果锥虫特异性DHS 与我们计划的提案的R33相相比 至少具有适当特性的至少一个系列的铅来优化要提出的属性 帽子的处理。识别的DHS抑制剂也将在命中验证期间在各个阶段进行测试 并针对T. Cruzi和Leishmania进行优化，以确定它们是否有潜力 开发为所有三种基于锥虫的疾病的广谱治疗方法。