Identification of Novel Plasmodium falciparum Hexokinase Inhibitors with Antiparasitic Activity.

Abstract:

:Plasmodium falciparum, the deadliest species of malaria parasites, is dependent on glycolysis for the generation of ATP during the pathogenic red blood cell stage. Hexokinase (HK) catalyzes the first step in glycolysis, transferring the γ-phosphoryl group of ATP to glucose to yield glucose-6-phosphate. Here, we describe the validation of a high-throughput assay for screening small-molecule collections to identify inhibitors of the P. falciparum HK (PfHK). The assay, which employed an ADP-Glo reporter system in a 1,536-well-plate format, was robust with a signal-to-background ratio of 3.4 ± 1.2, a coefficient of variation of 6.8% ± 2.9%, and a Z'-factor of 0.75 ± 0.08. Using this assay, we screened 57,654 molecules from multiple small-molecule collections. Confirmed hits were resolved into four clusters on the basis of structural relatedness. Multiple singleton hits were also identified. The most potent inhibitors had 50% inhibitory concentrations as low as ∼1 μM, and several were found to have low-micromolar 50% effective concentrations against asexual intraerythrocytic-stage P. falciparum parasites. These molecules additionally demonstrated limited toxicity against a panel of mammalian cells. The identification of PfHK inhibitors with antiparasitic activity using this validated screening assay is encouraging, as it justifies additional HTS campaigns with more structurally amenable libraries for the identification of potential leads for future therapeutic development.

authors

Davis MI,Patrick SL,Blanding WM,Dwivedi V,Suryadi J,Golden JE,Coussens NP,Lee OW,Shen M,Boxer MB,Hall MD,Sharlow ER,Drew ME,Morris JC

doi

10.1128/AAC.00914-16

subject

Has Abstract

pub_date

2016-09-23 00:00:00

pages

6023-33

issue

10

eissn

0066-4804

issn

1098-6596

pii

AAC.00914-16

journal_volume

60

pub_type

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