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.

项目摘要 人类非洲锥虫症（HAT）是由各种亚种引起的贫困疾病 布鲁氏锥虫原生动物寄生虫。 TSE-TSE苍蝇的咬伤传播，第1期疾病是 以通过血液和淋巴系统传播的锥虫特征，然后越过 血脑屏障进入中枢神经系统，引起2期疾病。中枢神经系统的渗透与令人恐惧的 一系列逐渐恶化，最终致命的心理和生理障碍。药品 治疗依赖于必须在医疗监督下进行的过时且经常有毒药物 建立了哪些阻力或持续的威胁。需要新药。蛋白酶体是一种进化 - 在细胞中保守的多生产蛋白复合物调节正常蛋白质周转和降解的细胞中 错误折叠的蛋白质。用小分子抑制剂抑制t. brucei蛋白酶体（TB20）杀死 锥虫。但是，关于蛋白水解裂解偏好或 TB20S目标的抑制作用。发现这些信息将支持开发 蛋白酶体抑制剂具有提高的特异性和效力，这可能构成新处理的基础 帽子。因此，在AIM 1中，我们将使用A全面介绍TB20的裂解特异性 通过质谱法（MSP-MS）称为多重底物分析的技术。产生的数据将告知（i） 可以测量TB20活性的优化肽基底物的合成和（ii）A 药物化学努力（目标3）以提高特异性，合成新的TB20抑制剂。在AIM 2中，我们 将用各种蛋白酶体抑制剂库筛选TB20，其中包含不同的支架和反应性群 由内部和合作者开发的。产生的数据将告知（i）更广泛的化学物质 与抑制TB20和（ii）抑制有关的空间，同样计划了AIM 3的药物化学计划。 对于AIM 3，我们将筛选Brucei锥虫瘤和哺乳动物细胞系，具有最高的TB20 抑制剂了解寄生虫杀死和细胞毒性之间的差异。这个目标也涉及 集中的药物化学运动，评估所有蛋白酶体抑制和细胞筛选数据 使用称为海洋天然产物的脚手架，合成具有提高特异性的TB20抑制剂 Carmaphycin B.拟议的R21研究完成后，我们将确定一系列TB20S 抑制剂准备进一步发展作为帽子的治疗学。