Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development

Roman Deniskin, I. J. Frame, Yvett Sosa, Myles Akabas

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Infection with Plasmodium falciparum and vivax cause most cases of malaria. Emerging resistance to current antimalarial medications makes new drug development imperative. Ideally a new antimalarial drug should treat both falciparum and vivax malaria. Because malaria parasites are purine auxotrophic, they rely on purines imported from the host erythrocyte via Equilibrative Nucleoside Transporters (ENTs). Thus, the purine import transporters represent a potential target for antimalarial drug development. For falciparum parasites the primary purine transporter is the P. falciparum Equilibrative Nucleoside Transporter Type 1 (PfENT1). Recently we identified potent PfENT1 inhibitors with nanomolar IC50 values using a robust, yeast-based high throughput screening assay. In the current work we characterized the Plasmodium vivax ENT1 (PvENT1) homologue and its sensitivity to the PfENT1 inhibitors. We expressed a yeast codon-optimized PvENT1 gene in Saccharomyces cerevisiae. PvENT1-expressing yeast imported both purines ([3H]adenosine) and pyrimidines ([3H]uridine), whereas wild type (fui1δ) yeast did not. Based on radiolabel substrate uptake inhibition experiments, inosine had the lowest IC50 (3.8 μM), compared to guanosine (14.9 μM) and adenosine (142 μM). For pyrimidines, thymidine had an IC50 of 183 μM (vs. cytidine and uridine; mM range). IC50 values were higher for nucleobases compared to the corresponding nucleosides; hypoxanthine had a 25-fold higher IC50 than inosine. The archetypal human ENT1 inhibitor 4-nitrobenzylthioinosine (NBMPR) had no effect on PvENT1, whereas dipyridamole inhibited PvENT1, albeit with a 40 μM IC50, a 1000-fold less sensitive than human ENT1 (hENT1). The PfENT1 inhibitors blocked transport activity of PvENT1 and the five known naturally occurring non-synonymous single nucleotide polymorphisms (SNPs) with similar IC50 values. Thus, the PfENT1 inhibitors also target PvENT1. This implies that development of novel antimalarial drugs that target both falciparum and vivax ENT1 may be feasible.

Original languageEnglish (US)
Pages (from-to)1-11
Number of pages11
JournalInternational Journal for Parasitology: Drugs and Drug Resistance
Volume6
Issue number1
DOIs
StatePublished - Apr 1 2016

Fingerprint

Equilibrative Nucleoside Transporter 1
Plasmodium vivax
Antimalarials
Inhibitory Concentration 50
Plasmodium falciparum
Yeasts
Inosine
Pyrimidines
Purines
Uridine
Adenosine
Malaria
Parasites
Nucleoside Transport Proteins
High-Throughput Screening Assays
Vivax Malaria
Cytidine
Hypoxanthine
Dipyridamole
Falciparum Malaria

Keywords

  • Drug development
  • Malaria
  • Nucleoside/nucleobase transport
  • Parasite
  • Plasmodium vivax
  • Purines
  • Single-nucleotide polymorphism (SNP)
  • Transporter

ASJC Scopus subject areas

  • Infectious Diseases
  • Pharmacology (medical)
  • Pharmacology

Cite this

Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development. / Deniskin, Roman; Frame, I. J.; Sosa, Yvett; Akabas, Myles.

In: International Journal for Parasitology: Drugs and Drug Resistance, Vol. 6, No. 1, 01.04.2016, p. 1-11.

Research output: Contribution to journalArticle

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