Hijacking Plasmodium ubiquitin-proteasome system to defeat drug resistance

劫持疟原虫泛素蛋白酶体系统以击败耐药性

• 批准号: 10719157
• 负责人: Gang Lin
• 金额: $ 75.27万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719157
• 关键词: Active Sites; Africa; African; Antimalarials; Artemisinins; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Availability; Cells; Cessation of life; Chemicals; Child; Clinical; Combined Modality Therapy; Communicable Diseases; Distal; Drug Combinations; Drug Kinetics; Drug resistance; Exposure to; Frequencies; Goals; HIV; Hemin; Hybrids; In Vitro; Infection; Investigation; Life Cycle Stages; Lipids; Malaria; Malignant Neoplasms; Modeling; Multienzyme Complexes; Mus; Mutation; Oligopeptides; Oral; Organism; Parasite resistance; Parasites; Penetration; Peptides; Pharmaceutical Preparations; Plasmodium; Plasmodium falciparum; Point Mutation; Prevention; Process; Prodrugs; Property; Proteasome Inhibitor; Proteins; Recrudescences; Reporting; Resistance; Resistance development; Site; Southeastern Asia; Structure-Activity Relationship; System; Testing; Therapeutic; Time; Tissues; Toxic effect; Ubiquitin; Vertebral column; analog; cytotoxicity; detection limit; efficacy study; efficacy testing; global health; high risk; humanized mouse; improved; in vivo; inhibitor; lead optimization; mouse model; multicatalytic endopeptidase complex; multidrug tolerance; mutant; novel; oxidative damage; pharmacophore; prevent; resistance mechanism; resistance mutation; standard care; success; synergism; tuberculosis treatment; Active Sites; Africa; African; Antimalarials; Artemisinins; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Availability; Cells; Cessation of life; Chemicals; Child; Clinical; Combined Modality Therapy; Communicable Diseases; Distal; Drug Combinations; Drug Kinetics; Drug resistance; Exposure to; Frequencies; Goals; HIV; Hemin; Hybrids; In Vitro; Infection; Investigation; Life Cycle Stages; Lipids; Malaria; Malignant Neoplasms; Modeling; Multienzyme Complexes; Mus; Mutation; Oligopeptides; Oral; Organism; Parasite resistance; Parasites; Penetration; Peptides; Pharmaceutical Preparations; Plasmodium; Plasmodium falciparum; Point Mutation; Prevention; Process; Prodrugs; Property; Proteasome Inhibitor; Proteins; Recrudescences; Reporting; Resistance; Resistance development; Site; Southeastern Asia; Structure-Activity Relationship; System; Testing; Therapeutic; Time; Tissues; Toxic effect; Ubiquitin; Vertebral column; analog; cytotoxicity; detection limit; efficacy study; efficacy testing; global health; high risk; humanized mouse; improved; in vivo; inhibitor; lead optimization; mouse model; multicatalytic endopeptidase complex; multidrug tolerance; mutant; novel; oxidative damage; pharmacophore; prevent; resistance mechanism; resistance mutation; standard care; success; synergism; tuberculosis treatment

Project Summary/Abstract In 2020, with over 250 million debilitating cases and over half a million deaths, mostly in young children, malaria is a persistent global health crisis. The malaria-causing parasite Plasmodium falciparum (Pf) has developed resistance to most antimalarial drug deployed, including the backbone artemisinins (ARTs). ART and its semi-synthetic analogs are considered essential for malaria treatment. ARTs are prodrugs that are activated within the parasites to form a reactive radical that covalently attacks proteins, lipids and other cellular constituents. ART resistance is widespread in Southeast Asia and has been reported in Africa. ART combination therapy (ACT) is a mainstay for treatment of malaria, but its efficacy can be derailed when a two-drug combination becomes de facto monotherapy. Moreover, extended exposure of Pf to ACTs induces multidrug tolerance. We recently showed that inhibitors specific for the Pf proteasome (Pf20S) kill Pf in each stage of its life cycle and synergize with ART, overcoming ART resistance. This proposal builds on our discovery that a covalent hybrid of an ART analogue and a Pf20S inhibitor that we call an artezomib (ATZ) can enhance ART action and overcome resistance to each of its components. We have synthesized ATZs that are more potent Pf20S inhibitors than their component Pf20S inhibitor. They not only kill wild type and ART-resistant (K13 mutant) Pf, Pf with proteasome mutations that confer resistance to the Pf20S inhibitor, but also kill Pf that expresses both ART-resistant and PI-resistant mutations. We propose the following mechanism by which ATZs overcome resistance to the Pf20S inhibitor within them: We found that upon activation of ATZ in the parasites, the ART component binds Pf proteins, like activated ART itself. The Pf ubiquitin proteasome system digests ATZ-bound proteins into oligopeptides, some of which display the Pf inhibitor component of the ATZ. We hypothesize that extended contact of ATZ-bearing peptides within the Pf20S active site augments the binding of the Pf20S inhibitor component of the ATZ, overcoming the decreased binding otherwise conferred by Pf20S point mutations. Thus, an ATZ can overcome resistance to each of its components. In mouse models of malaria, an ATZ drove P. berghei below the limit of detection and suppressed recrudescence of a P. berghei ART-resistant K13 mutant and doing so better than ART. In Aim 1 of this proposal, we will conduct lead optimization to improve ATZs' potency, selectivity and ATZs' pharmacokinetic properties. In Aim 2, we will explore ATZs' mechanism of action; attempt to select for ATZ-resistant parasites; determine the frequency and mechanism of resistance, if any; and study antimalarial activity of ATZs in stages of the Pf life cycle when ART alone is ineffective. Aim 3 will test the efficacy of ATZs in mice, including humanized mice infected with Pf. .

项目摘要/摘要 在2020年，超过2.5亿个使人衰弱的案件和超过半百万的死亡，主要是幼儿， 疟疾是一场持续的全球健康危机。引起疟疾的寄生虫疟原虫恶性疟原虫（PF） 已经对部署的大多数抗疟药产生了抗药性，包括骨链蛋白蛋白蛋白 （艺术）。艺术及其半合成类似物被认为是疟疾治疗必不可少的。艺术是 在寄生虫中激活的前药形成一个共价攻击的反应性自由基 蛋白质，脂质和其他细胞成分。艺术抵抗在东南亚广泛，有 在非洲报道。艺术联合疗法（ACT）是治疗疟疾的中流阶段，但 当两种药物组合成为事实上的单一疗法时，疗效可能会脱轨。而且， PF对行为的延长暴露会诱导多药公差。我们最近表明抑制剂 特定于PF蛋白酶体（PF20）在其生命周期的每个阶段杀死PF，并与艺术协同作用， 克服艺术的抵抗力。这项提议是基于我们发现的，即艺术的共价杂种 我们称为Artezomib（ATZ）的模拟和PF20抑制剂可以增强艺术作用并克服 对每个组件的抗性。我们拥有更有效的PF20抑制剂的合成ATZ 比它们的成分PF20抑制剂。他们不仅杀死野生型和抗艺术（K13突变体）PF，PF 蛋白酶体突变赋予PF20抑制剂的抗性，但也杀死表达的PF 抗艺术和耐Pi的突变。我们提出了以下ATZ的机制 克服对其中PF20抑制剂的抗性：我们发现在激活ATZ时 寄生虫，艺术成分结合了PF蛋白，就像活化的艺术本身一样。 PF泛素蛋白酶体 系统消化ATZ结合的蛋白进入寡肽，其中一些显示PF抑制剂 ATZ的组成部分。我们假设在 PF20活性位点增加了ATZ的PF20抑制剂成分的结合，克服了 PF20点突变赋予的结合减少。因此，ATZ可以克服 对每个组件的抗性。在疟疾的小鼠模型中，ATZ驱动了P. Berghei 抗伯格（P. 比艺术好。在本提案的目标1中，我们将进行铅优化以改善ATZ的 效力，选择性和ATZ的药代动力学特性。在AIM 2中，我们将探索ATZS的机制 行动；尝试选择抗ATZ的寄生虫；确定频率和机制 抵抗，如果有的话；并研究ATZ在PF生命周期阶段的ATZ的抗疟疾活性 无效。 AIM 3将测试ATZ在小鼠中的疗效，包括感染PF的人源化小鼠。 。