title: Metabolic Dysregulation Induced in Plasmodium falciparum by Dihydroartemisinin and Other Front-Line Antimalarial Drugs.
abstract: Detailed information on the mode of action of antimalarial drugs can be used to improve existing drugs, identify new drug targets, and understand the basis of drug resistance. In this study we describe the use of a time-resolved, mass spectrometry (MS)-based metabolite profiling approach to map the metabolic perturbations induced by a panel of clinical antimalarial drugs and inhibitors on Plasmodium falciparum asexual blood stages. Drug-induced changes in metabolite levels in P. falciparum-infected erythrocytes were monitored over time using gas chromatography-MS and liquid chromatography-MS and changes in specific metabolic fluxes confirmed by nonstationary [(13)C]-glucose labeling. Dihydroartemisinin (DHA) was found to disrupt hemoglobin catabolism within 1 hour of exposure, resulting in a transient decrease in hemoglobin-derived peptides. Unexpectedly, it also disrupted pyrimidine biosynthesis, resulting in increased [(13)C]-glucose flux toward malate production, potentially explaining the susceptibility of P. falciparum to DHA during early blood-stage development. Unique metabolic signatures were also found for atovaquone, chloroquine, proguanil, cycloguanil and methylene blue. We also show that this approach can be used to identify the mode of action of novel antimalarials, such as the compound Torin 2, which inhibits hemoglobin catabolism.
authors.affiliation: Department of Biochemistry and Molecular Biology.
authors.forename: Simon A
authors.initials: SA
authors.lastname: Cobbold
authors.affiliation: Department of Biochemistry and Molecular Biology.
authors.forename: Hwa H
authors.initials: HH
authors.lastname: Chua
authors.affiliation: Metabolomics Australia, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne.
authors.forename: Brunda
authors.initials: B
authors.lastname: Nijagal
authors.affiliation: Department of Biochemistry and Molecular Biology Drug Delivery Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
authors.forename: Darren J
authors.initials: DJ
authors.lastname: Creek
authors.affiliation: Department of Biochemistry and Molecular Biology.
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Ralph
authors.affiliation: Department of Biochemistry and Molecular Biology Metabolomics Australia, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne.
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: McConville
mesh_terms: D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology*; D053626:Atovaquone / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology; D005578:Fosfomycin / Q000031:analogs & derivatives / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D006454:Hemoglobins / Q000187:drug effects / Q000378:metabolism; D020128:Inhibitory Concentration 50; D008751:Methylene Blue / Q000494:pharmacology; D009287:Naphthyridines / Q000494:pharmacology; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000378:metabolism; D002727:Proguanil / Q000494:pharmacology; D014227:Triazines / Q000494:pharmacology
chemical_list: C558529:9-(6-aminopyridin-3-yl)-1-(3-(trifluoromethyl)phenyl)benzo(h)(1,6)naphthyridin-2(1H)-one; D000962:Antimalarials; D037621:Artemisinins; D006454:Hemoglobins; D009287:Naphthyridines; D014227:Triazines; C026009:cycloguanil; D005578:Fosfomycin; C024640:fosmidomycin; C039060:artenimol; D002738:Chloroquine; D002727:Proguanil; D008751:Methylene Blue; D053626:Atovaquone
keywords: Plasmodium; antimalarial; artemisinin; atovaquone; chloroquine; drug screening; malaria; metabolite; metabolomics; torin