Australian Defense Force Malaria and Infectious Disease Institute, Brisbane, Australia.
Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Australia.
Mol Cell Proteomics. 2020 Feb;19(2):308-325. doi: 10.1074/mcp.RA119.001797. Epub 2019 Dec 13.
The increasing incidence of antimalarial drug resistance to the first-line artemisinin combination therapies underpins an urgent need for new antimalarial drugs, ideally with a novel mode of action. The recently developed 2-aminomethylphenol, JPC-3210, (MMV 892646) is an erythrocytic schizonticide with potent antimalarial activity against multidrug-resistant lines, low cytotoxicity, potent efficacy against murine malaria, and favorable preclinical pharmacokinetics including a lengthy plasma elimination half-life. To investigate the impact of JPC-3210 on biochemical pathways within infected red blood cells, we have applied a "multi-omics" workflow based on high resolution orbitrap mass spectrometry combined with biochemical approaches. Metabolomics, peptidomics and hemoglobin fractionation analyses revealed a perturbation in hemoglobin metabolism following JPC-3210 exposure. The metabolomics data demonstrated a specific depletion of short hemoglobin-derived peptides, peptidomics analysis revealed a depletion of longer hemoglobin-derived peptides, and the hemoglobin fractionation assay demonstrated decreases in hemoglobin, heme and hemozoin levels. To further elucidate the mechanism responsible for inhibition of hemoglobin metabolism, we used β-hematin polymerization assays and showed JPC-3210 to be an intermediate inhibitor of β-hematin polymerization, about 10-fold less potent then the quinoline antimalarials, such as chloroquine and mefloquine. Further, quantitative proteomics analysis showed that JPC-3210 treatment results in a distinct proteomic signature compared with other known antimalarials. While JPC-3210 clustered closely with mefloquine in the metabolomics and proteomics analyses, a key differentiating signature for JPC-3210 was the significant enrichment of parasite proteins involved in regulation of translation. These studies revealed that the mode of action for JPC-3210 involves inhibition of the hemoglobin digestion pathway and elevation of regulators of protein translation. Importantly, JPC-3210 demonstrated rapid parasite killing kinetics compared with other quinolones, suggesting that JPC-3210 warrants further investigation as a potentially long acting partner drug for malaria treatment.
抗疟药耐药性的不断增加，使一线青蒿素联合疗法面临迫切需要新的抗疟药物，理想情况下是具有新作用模式的药物。最近开发的 2-氨甲基苯酚（JPC-3210，MMV892646）是一种红细胞裂殖体抑制剂，对多药耐药株具有强大的抗疟活性，细胞毒性低，对鼠疟具有强大的疗效，以及有利的临床前药代动力学，包括较长的血浆消除半衰期。为了研究 JPC-3210 对感染红细胞内生化途径的影响，我们应用了一种基于高分辨率轨道阱质谱结合生化方法的“多组学”工作流程。代谢组学、肽组学和血红蛋白分级分析显示，JPC-3210 暴露后血红蛋白代谢受到干扰。代谢组学数据表明，短血红蛋白衍生肽特异性耗竭，肽组学分析表明，长血红蛋白衍生肽耗竭，血红蛋白分级分析表明血红蛋白、血红素和血晶素水平降低。为了进一步阐明抑制血红蛋白代谢的机制，我们使用β-血红蛋白聚合测定法，表明 JPC-3210 是β-血红蛋白聚合的中间抑制剂，比氯喹和甲氟喹等喹啉类抗疟药的效力低约 10 倍。此外，定量蛋白质组学分析表明，JPC-3210 处理与其他已知抗疟药相比，导致明显不同的蛋白质组学特征。虽然 JPC-3210 在代谢组学和蛋白质组学分析中与甲氟喹聚类密切，但 JPC-3210 的一个关键区别特征是翻译调节相关寄生虫蛋白的显著富集。这些研究表明，JPC-3210 的作用模式涉及抑制血红蛋白消化途径和提高蛋白质翻译调节剂。重要的是，与其他喹啉类药物相比，JPC-3210 表现出快速的寄生虫杀伤动力学，这表明 JPC-3210 值得进一步研究，作为疟疾治疗的潜在长效联合药物。
