Inhibition of Hypusine Biosynthesis in Plasmodium: A Possible, New Strategy in Prevention and Therapy of Malaria

Kaiser, Annette; Ulmer, Daniela; Goebel, Tim; Holzgrabe, Ulrike; Saeftel, Michael; Hoerauf, Achim

doi:10.2174/138955706778742795

Cited by 13 publications

(5 citation statements)

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“…Multiple top ranking enzymes are involved in the process of protein biosynthesis (tRNA ligases, deoxyhypusine synthase). The eukaryotic initiation factor, whose activation involves deoxyhypusine synthase, has already been suggested to be the target of a drug with antimalarial effect [104]. Little is known about the translational machinery of P. falciparum despite the obvious global effect upon its inhibition.…”

Section: Resultsmentioning

confidence: 99%

Antimalarial drug targets in Plasmodium falciparum predicted by stage-specific metabolic network analysis

et al. 2010

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BackgroundDespite enormous efforts to combat malaria the disease still afflicts up to half a billion people each year of which more than one million die. Currently no approved vaccine is available and resistances to antimalarials are widely spread. Hence, new antimalarial drugs are urgently needed.ResultsHere, we present a computational analysis of the metabolism of Plasmodium falciparum, the deadliest malaria pathogen. We assembled a compartmentalized metabolic model and predicted life cycle stage specific metabolism with the help of a flux balance approach that integrates gene expression data. Predicted metabolite exchanges between parasite and host were found to be in good accordance with experimental findings when the parasite's metabolic network was embedded into that of its host (erythrocyte). Knock-out simulations identified 307 indispensable metabolic reactions within the parasite. 35 out of 57 experimentally demonstrated essential enzymes were recovered and another 16 enzymes, if additionally the assumption was made that nutrient uptake from the host cell is limited and all reactions catalyzed by the inhibited enzyme are blocked. This predicted set of putative drug targets, shown to be enriched with true targets by a factor of at least 2.75, was further analyzed with respect to homology to human enzymes, functional similarity to therapeutic targets in other organisms and their predicted potency for prophylaxis and disease treatment.ConclusionsThe results suggest that the set of essential enzymes predicted by our flux balance approach represents a promising starting point for further drug development.

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Section: Resultsmentioning

confidence: 99%

Antimalarial drug targets in Plasmodium falciparum predicted by stage-specific metabolic network analysis

et al. 2010

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“…Parasitic diseases are an enormous health problem, especially in many developing countries, requiring cost effective drugs. Fortunately, the polyamine metabolic pathway has been found to contain several potential drug targets in parasites (Heby et al 2007; Kaiser et al 2006; Muller et al 2008; Reguera et al 2005). Thus, targeting differences in polyamine metabolism between host and parasite, or healthy and malignant tissue may offer a selective advantage and avoid significant host toxicity.…”

Section: Introductionmentioning

confidence: 99%

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

et al. 2009

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SUMMARYN-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N, N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N'-(3-ethylaminopropyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD (K m(APAO) =10μM, k cat(APAO) =1.1s −1 and K m(SMO) =28μM, k cat(SMO) =0.8s −1 , respectively), metabolized BnEtSPM to EtSPD (K m(APAO) =0.9 μM, k cat(APAO) =1.1s −1 and K m(SMO) =51μM, k cat(SMO) =0.4s −1 , respectively), and metabolized DBSPM to BnSPD (K m(APAO) =5.4μM, k cat(APAO) = 2.0s −1 and K m(SMO) =33μM, k cat(SMO) =0.3s −1 , respectively). Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD (EtSPM K m(APAO) =16μM, k cat(APAO) =1.5s −1 ; K m(SMO) =25μM, k cat(SMO) =8.2s −1 ; BnSPM K m(APAO) =6.0μM, k cat(APAO) =2.8s −1 ; K m(SMO) =19μM, k cat(SMO) =0.8s −1 , respectively). Surprisingly, E t S P D ( K m(APAO) =37μM, k cat(APAO) =0.1s −1 ; K m(SMO) =48μM, k cat(SMO) =0.05s −1 ) and BnSPD (K m(APAO) =2.5μM, k cat(APAO) =3.5s −1 ; K m(SMO) =60μM, k cat(SMO) =0.54s −1 ) were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 hours of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could play an important role in polyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues.

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“…Data from mimosine, ciclopirox, 4-Oxo-piperidine-3-mono- carboxylate were taken from [40].*HUVEC = Human umbicilial Vein Endothelial Cells.…”

Section: Resultsmentioning

confidence: 99%

“…Another interesting observation derives from a comparison of different iron-chelating compounds (Table 2) [40] tested against the P. falciparum DOOH. These data show that zileuton inhibits DOHH from P. vivax in a nanomolar range while all the other compounds unfold inhibition at a micromolar concentration.…”

Section: Discussionmentioning

confidence: 99%

Deoxyhypusine Hydroxylase from Plasmodium vivax, the Neglected Human Malaria Parasite: Molecular Cloning, Expression and Specific Inhibition by the 5-LOX Inhibitor Zileuton

et al. 2013

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Primaquine, an 8-aminoquinoline, is the only drug which cures the dormant hypnozoites of persistent liver stages from P. vivax. Increasing resistance needs the discovery of alternative pathways as drug targets to develop novel drug entities. Deoxyhypusine hydroxylase (DOHH) completes hypusine biosynthesis in eukaryotic initiation factor (eIF-5A) which is the only cellular protein known to contain the unusual amino acid hypusine. Modified EIF-5A is important for proliferation of the malaria parasite. Here, we present the first successful cloning and expression of DOHH from P. vivax causing tertiary malaria. The nucleic acid sequence of 1041 bp encodes an open reading frame of 346 amino acids. Histidine tagged expression of P. vivax DOHH detected a protein of 39.01 kDa in E. coli. The DOHH protein from P. vivax shares significant amino acid identity to the simian orthologues from P. knowlesi and P. yoelii strain H. In contrast to P. falciparum only four E-Z-type HEAT-like repeats are present in P. vivax DOHH with different homology to phycocyanin lyase subunits from cyanobacteria and in proteins participating in energy metabolism of Archaea and Halobacteria. However, phycocyanin lyase activity is absent in P. vivax DOHH. The dohh gene is present as a single copy gene and transcribed throughout the whole erythrocytic cycle. Specific inhibition of recombinant P. vivax DOHH is possible by complexing the ferrous iron with zileuton, an inhibitor of mammalian 5-lipoxygenase (5-LOX). Ferrous iron in the active site of 5-LOX is coordinated by three conserved histidines and the carboxylate of isoleucine673. Zileuton inhibited the P. vivax DOHH protein with an IC50 of 12,5 nmol determined by a relative quantification by GC/MS. By contrast, the human orthologue is only less affected with an IC50 of 90 nmol suggesting a selective iron-complexing strategy for the parasitic enzyme.

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Inhibition of Hypusine Biosynthesis in Plasmodium: A Possible, New Strategy in Prevention and Therapy of Malaria

Cited by 13 publications

References 40 publications

Antimalarial drug targets in Plasmodium falciparum predicted by stage-specific metabolic network analysis

Antimalarial drug targets in Plasmodium falciparum predicted by stage-specific metabolic network analysis

Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases

Deoxyhypusine Hydroxylase from Plasmodium vivax, the Neglected Human Malaria Parasite: Molecular Cloning, Expression and Specific Inhibition by the 5-LOX Inhibitor Zileuton

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