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Pyrazoleamide compounds are potent antimalarials that target Na+ homeostasis in intraerythrocytic Plasmodium falciparum.

Vaidya AB, Morrisey JM, Zhang Z, Das S, Daly TM, Otto TD, Spillman NJ, Wyvratt M, Siegl P, Marfurt J, Wirjanata G, Sebayang BF, Price RN, Chatterjee A, Nagle A, Stasiak M, Charman SA, Angulo-Barturen I, Ferrer S, Belén Jiménez-Díaz M, Martínez MS, Gamo FJ, Avery VM, Ruecker A, Delves M, Kirk K, Berriman M, Kortagere S, Burrows J, Fan E, Bergman LW - Nat Commun (2014)

Bottom Line: Investigations involving pyrazoleamide-resistant parasites, whole-genome sequencing and gene transfers reveal that mutations in two proteins, a calcium-dependent protein kinase (PfCDPK5) and a P-type cation-ATPase (PfATP4), are necessary to impart full resistance to these compounds.A pyrazoleamide compound causes a rapid disruption of Na(+) regulation in blood-stage Plasmodium falciparum parasites.Similar effect on Na(+) homeostasis was recently reported for spiroindolones, which are antimalarials of a chemical class quite distinct from pyrazoleamides.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine, 2900 Queen Lane, Philadelphia, Pennsylvania 190129, USA.

ABSTRACT
The quest for new antimalarial drugs, especially those with novel modes of action, is essential in the face of emerging drug-resistant parasites. Here we describe a new chemical class of molecules, pyrazoleamides, with potent activity against human malaria parasites and showing remarkably rapid parasite clearance in an in vivo model. Investigations involving pyrazoleamide-resistant parasites, whole-genome sequencing and gene transfers reveal that mutations in two proteins, a calcium-dependent protein kinase (PfCDPK5) and a P-type cation-ATPase (PfATP4), are necessary to impart full resistance to these compounds. A pyrazoleamide compound causes a rapid disruption of Na(+) regulation in blood-stage Plasmodium falciparum parasites. Similar effect on Na(+) homeostasis was recently reported for spiroindolones, which are antimalarials of a chemical class quite distinct from pyrazoleamides. Our results reveal that disruption of Na(+) homeostasis in malaria parasites is a promising mode of antimalarial action mediated by at least two distinct chemical classes.

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Mutations in compound C2-1-resistant parasites.A Thr to Ala mutation at position 392 in PfCDPK5 is in its predicted junction domain (a), and a Val to Ile mutation at position 178 in PfATP4 (b) is in the first predicted transmembrane domain.
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f3: Mutations in compound C2-1-resistant parasites.A Thr to Ala mutation at position 392 in PfCDPK5 is in its predicted junction domain (a), and a Val to Ile mutation at position 178 in PfATP4 (b) is in the first predicted transmembrane domain.

Mentions: To gain insight into the mode of action for and resistance to the pyrazoleamides, genomes of the Dd2 parental line and the three pyrazoleamide-resistant lines were sequenced using ‘next-generation’ sequencing technology17. Custom-designed bioinformatics for P. falciparum genome-sequence comparison18 revealed non-synonymous single-nucleotide polymorphism (SNP) in five genes: PF3D7_0411900, DNA polymerase alpha (mutation D133Y); PF3D7_0630900, DEAD/DEAH box ATP-dependent RNA helicase (N142K, a polymorphic site); PF3D7_1034300, thioredoxin-like associated protein 2, (K465E); PF3D7_1211900, non-SERCA calcium ATPase (V178I); and PF3D7_1337800, calcium-dependent protein kinase 5 (T392A). These mutations were common to all three resistant lines compared with the parental Dd2 parasites. An assessment of potential effects of these mutations as well as expression patterns of these genes led us to focus initial attention on two proteins: the calcium-dependent protein kinase 5 (PfCDPK5; PF3D7_1337800) and a P-type cation ATPase annotated as a non-SERCA calcium ATPase, PfATP4 (PF3D7_1211900). Dvorin et al.19 have shown PfCDPK5 to be essential for egress of progeny merozoites at the last step of erythrocytic schizogony by P. falciparum, and thus deemed an attractive drug target. The resistance-associated mutation, T392A, in PfCDPK5 was predicted to be in the junction region of the enzyme that links the kinase domain to the calmodulin-like EF-hand domains, and acts as a regulatory loop of the enzyme (Fig. 3a). PfATP4 was first described as a Ca2+ pump2021 and recently found to bear mutations in P. falciparum parasites resistant to a new class of antimalarials, the spiroindolones9. Spillman et al.10 have provided evidence consistent with the hypothesis that PfATP4 is a P-type Na+ ATPase and a target for the spiroindolones. The resistance-associated mutation in PfATP4, a conservative V178I change, was localized to the predicted first transmembrane domain of the protein (Fig. 3b), and was different from the mutations observed by Rottmann et al.9 in spiroindolone-resistant parasites. The pyrazoleamide-resistant lines described here were not cross-resistant to a spiroindolone (Supplementary Fig. 3b).


Pyrazoleamide compounds are potent antimalarials that target Na+ homeostasis in intraerythrocytic Plasmodium falciparum.

Vaidya AB, Morrisey JM, Zhang Z, Das S, Daly TM, Otto TD, Spillman NJ, Wyvratt M, Siegl P, Marfurt J, Wirjanata G, Sebayang BF, Price RN, Chatterjee A, Nagle A, Stasiak M, Charman SA, Angulo-Barturen I, Ferrer S, Belén Jiménez-Díaz M, Martínez MS, Gamo FJ, Avery VM, Ruecker A, Delves M, Kirk K, Berriman M, Kortagere S, Burrows J, Fan E, Bergman LW - Nat Commun (2014)

Mutations in compound C2-1-resistant parasites.A Thr to Ala mutation at position 392 in PfCDPK5 is in its predicted junction domain (a), and a Val to Ile mutation at position 178 in PfATP4 (b) is in the first predicted transmembrane domain.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263321&req=5

f3: Mutations in compound C2-1-resistant parasites.A Thr to Ala mutation at position 392 in PfCDPK5 is in its predicted junction domain (a), and a Val to Ile mutation at position 178 in PfATP4 (b) is in the first predicted transmembrane domain.
Mentions: To gain insight into the mode of action for and resistance to the pyrazoleamides, genomes of the Dd2 parental line and the three pyrazoleamide-resistant lines were sequenced using ‘next-generation’ sequencing technology17. Custom-designed bioinformatics for P. falciparum genome-sequence comparison18 revealed non-synonymous single-nucleotide polymorphism (SNP) in five genes: PF3D7_0411900, DNA polymerase alpha (mutation D133Y); PF3D7_0630900, DEAD/DEAH box ATP-dependent RNA helicase (N142K, a polymorphic site); PF3D7_1034300, thioredoxin-like associated protein 2, (K465E); PF3D7_1211900, non-SERCA calcium ATPase (V178I); and PF3D7_1337800, calcium-dependent protein kinase 5 (T392A). These mutations were common to all three resistant lines compared with the parental Dd2 parasites. An assessment of potential effects of these mutations as well as expression patterns of these genes led us to focus initial attention on two proteins: the calcium-dependent protein kinase 5 (PfCDPK5; PF3D7_1337800) and a P-type cation ATPase annotated as a non-SERCA calcium ATPase, PfATP4 (PF3D7_1211900). Dvorin et al.19 have shown PfCDPK5 to be essential for egress of progeny merozoites at the last step of erythrocytic schizogony by P. falciparum, and thus deemed an attractive drug target. The resistance-associated mutation, T392A, in PfCDPK5 was predicted to be in the junction region of the enzyme that links the kinase domain to the calmodulin-like EF-hand domains, and acts as a regulatory loop of the enzyme (Fig. 3a). PfATP4 was first described as a Ca2+ pump2021 and recently found to bear mutations in P. falciparum parasites resistant to a new class of antimalarials, the spiroindolones9. Spillman et al.10 have provided evidence consistent with the hypothesis that PfATP4 is a P-type Na+ ATPase and a target for the spiroindolones. The resistance-associated mutation in PfATP4, a conservative V178I change, was localized to the predicted first transmembrane domain of the protein (Fig. 3b), and was different from the mutations observed by Rottmann et al.9 in spiroindolone-resistant parasites. The pyrazoleamide-resistant lines described here were not cross-resistant to a spiroindolone (Supplementary Fig. 3b).

Bottom Line: Investigations involving pyrazoleamide-resistant parasites, whole-genome sequencing and gene transfers reveal that mutations in two proteins, a calcium-dependent protein kinase (PfCDPK5) and a P-type cation-ATPase (PfATP4), are necessary to impart full resistance to these compounds.A pyrazoleamide compound causes a rapid disruption of Na(+) regulation in blood-stage Plasmodium falciparum parasites.Similar effect on Na(+) homeostasis was recently reported for spiroindolones, which are antimalarials of a chemical class quite distinct from pyrazoleamides.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine, 2900 Queen Lane, Philadelphia, Pennsylvania 190129, USA.

ABSTRACT
The quest for new antimalarial drugs, especially those with novel modes of action, is essential in the face of emerging drug-resistant parasites. Here we describe a new chemical class of molecules, pyrazoleamides, with potent activity against human malaria parasites and showing remarkably rapid parasite clearance in an in vivo model. Investigations involving pyrazoleamide-resistant parasites, whole-genome sequencing and gene transfers reveal that mutations in two proteins, a calcium-dependent protein kinase (PfCDPK5) and a P-type cation-ATPase (PfATP4), are necessary to impart full resistance to these compounds. A pyrazoleamide compound causes a rapid disruption of Na(+) regulation in blood-stage Plasmodium falciparum parasites. Similar effect on Na(+) homeostasis was recently reported for spiroindolones, which are antimalarials of a chemical class quite distinct from pyrazoleamides. Our results reveal that disruption of Na(+) homeostasis in malaria parasites is a promising mode of antimalarial action mediated by at least two distinct chemical classes.

Show MeSH
Related in: MedlinePlus