The catalytic core of the proteasome as a drug target to treat Human African Trypanosomiasis

蛋白酶体的催化核心作为治疗非洲人类锥虫病的药物靶点

• 批准号: 10677879
• 负责人: Conor Caffrey
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10677879
• 关键词: Adverse event; African; African Trypanosomiasis; Architecture; Binding; Bite; Blood; Caspase; Catalytic Domain; Cell Line; Cells; Central Nervous System; Chemicals; Collection; Complex; Data; Deterioration; Development; Disease; Drug Targeting; Eukaryota; Human; Hydrolysis; In Vitro; Industrialization; Infiltration; Invaded; Ketones; Knowledge; Leishmania; Libraries; Life; Lymphatic System; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Measures; Medical; Parasites; Parasitic Diseases; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacotherapy; Physiological; Positioning Attribute; Poverty; Program Development; Property; Proteasome Inhibition; Proteasome Inhibitor; Protein Complex Subunit; Regimen; Research; Resistance; Series; Specificity; Structure; Substrate Specificity; Supervision; Technology; Testing; Therapeutic; Trypanocidal Agents; Trypanosoma; Trypanosoma brucei brucei; Trypanosoma cruzi; Trypsin; Tsetse Flies; Vaccines; alternative treatment; blood-brain barrier crossing; chymotrypsin; cytotoxicity; design; expectation; improved; inhibitor; marine natural product; misfolded protein; multicatalytic endopeptidase complex; novel; novel therapeutics; parenteral administration; preclinical development; preference; programs; protein complex; protein degradation; psychologic; rational design; scaffold; screening; small molecule; small molecule inhibitor; transmission process

PROJECT SUMMARY Human African Trypanosomiases (HAT) is a disease of poverty caused by various sub-species of the Trypanosoma brucei protozoan parasite. Transmitted by the bite of the tse-tse fly, Stage 1 disease is characterized by trypanosomes that disseminate through the blood and lymphatic systems, and then cross the blood-brain barrier into the CNS to cause Stage 2 disease. Infiltration of the CNS is associated with a frightening array of progressively deteriorating and eventually lethal psychological and physiological disorders. Drug therapy relies on antiquated and often toxic drugs that must be administered under medical supervision and for which resistance is established or a constant threat. New drugs are needed. The proteasome is an evolutionarily- conserved, multi-subunit protein complex in the cell that regulates normal protein turnover and degradation of misfolded proteins. Inhibition of the T. brucei proteasome (Tb20S) with small molecule inhibitors kills trypanosomes. However, there is little information regarding the proteolytic cleavage preferences or the inhibition profile of the Tb20S target. Uncovering this information would support the development of proteasome inhibitors with improved specificity and potency that could then form the basis for a new treatment of HAT. Accordingly, in Aim 1 we will comprehensively profile the cleavage specificity of Tb20S using a technology called multiplex substrate profiling by mass spectrometry (MSP-MS). The data arising will inform (i) the synthesis of optimized peptidyl substrates with which the activity of Tb20S can be measured and (ii) a medicinal chemistry effort (Aim 3) to synthesize new Tb20S inhibitors with improved specificity. In Aim 2, we will screen Tb20S with various proteasome inhibitor libraries containing diverse scaffolds and reactive groups that have been developed in-house and by collaborators. The data arising will inform (i) the broader chemical space associated with inhibition of Tb20S and (ii), again, the medicinal chemistry program planned for Aim 3. For Aim 3, we will screen both Trypanosoma brucei and mammalian cell lines with the top performing Tb20S inhibitors to understand the differential between parasite-killing and cytotoxicity. This Aim also involves a focused medicinal chemistry campaign that evaluates all of the proteasome inhibition and cell screening data to synthesize Tb20S inhibitors with improved specificity using the scaffold of the marine natural product known as carmaphycin B. Upon completion of the proposed R21 studies, we will have identified a series of Tb20S inhibitors ready for further development as therapeutics for HAT.

项目概要 非洲人类锥虫病 (HAT) 是一种由非洲锥虫亚种引起的贫困疾病。 布氏锥虫原生动物寄生虫。通过采采蝇叮咬传播，第一阶段疾病是 其特点是锥虫通过血液和淋巴系统传播，然后穿过 血脑屏障进入中枢神经系统，导致第二阶段疾病。中枢神经系统的浸润与可怕的 一系列逐渐恶化并最终致命的心理和生理疾病。药品 治疗依赖于陈旧且通常有毒的药物，这些药物必须在医疗监督下使用 哪种抵抗是建立的还是一种持续的威胁。需要新药。蛋白酶体是一种进化- 细胞中保守的多亚基蛋白质复合物，调节正常的蛋白质周转和降解 错误折叠的蛋白质。用小分子抑制剂抑制布氏锥虫蛋白酶体 (Tb20S) 可导致死亡 锥虫。然而，关于蛋白水解切割偏好或 Tb20S 靶标的抑制曲线。发现这些信息将有助于开发 具有改进的特异性和效力的蛋白酶体抑制剂，可以为新的治疗奠定基础 帽子。因此，在目标 1 中，我们将使用 称为多重底物质谱分析 (MSP-MS) 的技术。产生的数据将告知 (i) 优化肽基底物的合成，用其可以测量 Tb20S 的活性，以及​​ (ii) 药物化学努力（目标 3）合成具有更高特异性的新型 Tb20S 抑制剂。在目标 2 中，我们 将使用包含不同支架和反应基团的各种蛋白酶体抑制剂库筛选 Tb20S 由内部和合作者开发。产生的数据将告知 (i) 更广泛的化学品 与 Tb20S 抑制相关的空间，以及 (ii) 为目标 3 规划的药物化学计划。 对于目标 3，我们将筛选具有最佳性能的 Tb20S 布氏锥虫和哺乳动物细胞系 抑制剂来了解寄生虫杀灭和细胞毒性之间的差异。这一目标还涉及 重点药物化学活动，评估所有蛋白酶体抑制和细胞筛选数据 使用海洋天然产物的支架合成具有更高特异性的 Tb20S 抑制剂 卡马霉素 B。完成拟议的 R21 研究后，我们将鉴定出一系列 Tb20S 抑制剂已准备好进一步开发为 HAT 疗法。