Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids

Diego Benítez, Andrea Medeiros, Lucía Fiestas, Esteban A. Panozzo-Zenere, Franziska Maiwald, Kyriakos C. Prousis, Marina Roussaki, Theodora Calogeropoulou, Anastasia Detsi, Timo Jaeger, Jonas Šarlauskas, Lucíja Peterlin Mašič, Conrad Kunick, Guillermo R. Labadie, Leopold Flohé, Marcelo A. Comini

Research output: Contribution to journalArticle

Abstract

Background: The search for novel chemical entities targeting essential and parasite-specific pathways is considered a priority for neglected diseases such as trypanosomiasis and leishmaniasis. The thiol-dependent redox metabolism of trypanosomatids relies on bis-glutathionylspermidine [trypanothione, T(SH)2], a low molecular mass cosubstrate absent in the host. In pathogenic trypanosomatids, a single enzyme, trypanothione synthetase (TryS), catalyzes trypanothione biosynthesis, which is indispensable for parasite survival. Thus, TryS qualifies as an attractive drug target candidate. Methodology/Principal Finding: A library composed of 144 compounds from 7 different families and several singletons was screened against TryS from three major pathogen species (Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum). The screening conditions were adjusted to the TryS´ kinetic parameters and intracellular concentration of substrates corresponding to each trypanosomatid species, and/or to avoid assay interference. The screening assay yielded suitable Z’ and signal to noise values (≥0.85 and ~3.5, respectively), and high intra-assay reproducibility. Several novel chemical scaffolds were identified as low μM and selective tri-tryp TryS inhibitors. Compounds displaying multi-TryS inhibition (N,N'-bis(3,4-substituted-benzyl) diamine derivatives) and an N5-substituted paullone (MOL2008) halted the proliferation of infective Trypanosoma brucei (EC50 in the nM range) and Leishmania infantum promastigotes (EC50 = 12 μM), respectively. A bis-benzyl diamine derivative and MOL2008 depleted intracellular trypanothione in treated parasites, which confirmed the on-target activity of these compounds. Conclusions/Significance: Novel molecular scaffolds with on-target mode of action were identified as hit candidates for TryS inhibition. Due to the remarkable species-specificity exhibited by tri-tryp TryS towards the compounds, future optimization and screening campaigns should aim at designing and detecting, respectively, more potent and broad-range TryS inhibitors.

Original languageEnglish (US)
Article numbere0004617
JournalPLoS Neglected Tropical Diseases
Volume10
Issue number4
DOIs
StatePublished - Apr 12 2016
Externally publishedYes

Fingerprint

trypanothione synthetase
Leishmania infantum
Trypanosoma brucei brucei
Parasites
Neglected Diseases
Species Specificity
Diamines
Leishmaniasis
Trypanosomiasis
Trypanosoma cruzi

ASJC Scopus subject areas

  • Infectious Diseases
  • Public Health, Environmental and Occupational Health
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Benítez, D., Medeiros, A., Fiestas, L., Panozzo-Zenere, E. A., Maiwald, F., Prousis, K. C., ... Comini, M. A. (2016). Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids. PLoS Neglected Tropical Diseases, 10(4), [e0004617]. https://doi.org/10.1371/journal.pntd.0004617

Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids. / Benítez, Diego; Medeiros, Andrea; Fiestas, Lucía; Panozzo-Zenere, Esteban A.; Maiwald, Franziska; Prousis, Kyriakos C.; Roussaki, Marina; Calogeropoulou, Theodora; Detsi, Anastasia; Jaeger, Timo; Šarlauskas, Jonas; Peterlin Mašič, Lucíja; Kunick, Conrad; Labadie, Guillermo R.; Flohé, Leopold; Comini, Marcelo A.

In: PLoS Neglected Tropical Diseases, Vol. 10, No. 4, e0004617, 12.04.2016.

Research output: Contribution to journalArticle

Benítez, D, Medeiros, A, Fiestas, L, Panozzo-Zenere, EA, Maiwald, F, Prousis, KC, Roussaki, M, Calogeropoulou, T, Detsi, A, Jaeger, T, Šarlauskas, J, Peterlin Mašič, L, Kunick, C, Labadie, GR, Flohé, L & Comini, MA 2016, 'Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids', PLoS Neglected Tropical Diseases, vol. 10, no. 4, e0004617. https://doi.org/10.1371/journal.pntd.0004617
Benítez, Diego ; Medeiros, Andrea ; Fiestas, Lucía ; Panozzo-Zenere, Esteban A. ; Maiwald, Franziska ; Prousis, Kyriakos C. ; Roussaki, Marina ; Calogeropoulou, Theodora ; Detsi, Anastasia ; Jaeger, Timo ; Šarlauskas, Jonas ; Peterlin Mašič, Lucíja ; Kunick, Conrad ; Labadie, Guillermo R. ; Flohé, Leopold ; Comini, Marcelo A. / Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids. In: PLoS Neglected Tropical Diseases. 2016 ; Vol. 10, No. 4.
@article{7cf6d07871d5420e8515ce38cf2cbccf,
title = "Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids",
abstract = "Background: The search for novel chemical entities targeting essential and parasite-specific pathways is considered a priority for neglected diseases such as trypanosomiasis and leishmaniasis. The thiol-dependent redox metabolism of trypanosomatids relies on bis-glutathionylspermidine [trypanothione, T(SH)2], a low molecular mass cosubstrate absent in the host. In pathogenic trypanosomatids, a single enzyme, trypanothione synthetase (TryS), catalyzes trypanothione biosynthesis, which is indispensable for parasite survival. Thus, TryS qualifies as an attractive drug target candidate. Methodology/Principal Finding: A library composed of 144 compounds from 7 different families and several singletons was screened against TryS from three major pathogen species (Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum). The screening conditions were adjusted to the TryS´ kinetic parameters and intracellular concentration of substrates corresponding to each trypanosomatid species, and/or to avoid assay interference. The screening assay yielded suitable Z’ and signal to noise values (≥0.85 and ~3.5, respectively), and high intra-assay reproducibility. Several novel chemical scaffolds were identified as low μM and selective tri-tryp TryS inhibitors. Compounds displaying multi-TryS inhibition (N,N'-bis(3,4-substituted-benzyl) diamine derivatives) and an N5-substituted paullone (MOL2008) halted the proliferation of infective Trypanosoma brucei (EC50 in the nM range) and Leishmania infantum promastigotes (EC50 = 12 μM), respectively. A bis-benzyl diamine derivative and MOL2008 depleted intracellular trypanothione in treated parasites, which confirmed the on-target activity of these compounds. Conclusions/Significance: Novel molecular scaffolds with on-target mode of action were identified as hit candidates for TryS inhibition. Due to the remarkable species-specificity exhibited by tri-tryp TryS towards the compounds, future optimization and screening campaigns should aim at designing and detecting, respectively, more potent and broad-range TryS inhibitors.",
author = "Diego Ben{\'i}tez and Andrea Medeiros and Luc{\'i}a Fiestas and Panozzo-Zenere, {Esteban A.} and Franziska Maiwald and Prousis, {Kyriakos C.} and Marina Roussaki and Theodora Calogeropoulou and Anastasia Detsi and Timo Jaeger and Jonas Šarlauskas and {Peterlin Mašič}, Luc{\'i}ja and Conrad Kunick and Labadie, {Guillermo R.} and Leopold Floh{\'e} and Comini, {Marcelo A.}",
year = "2016",
month = "4",
day = "12",
doi = "10.1371/journal.pntd.0004617",
language = "English (US)",
volume = "10",
journal = "PLoS Neglected Tropical Diseases",
issn = "1935-2727",
publisher = "Public Library of Science",
number = "4",

}

TY - JOUR

T1 - Identification of Novel Chemical Scaffolds Inhibiting Trypanothione Synthetase from Pathogenic Trypanosomatids

AU - Benítez, Diego

AU - Medeiros, Andrea

AU - Fiestas, Lucía

AU - Panozzo-Zenere, Esteban A.

AU - Maiwald, Franziska

AU - Prousis, Kyriakos C.

AU - Roussaki, Marina

AU - Calogeropoulou, Theodora

AU - Detsi, Anastasia

AU - Jaeger, Timo

AU - Šarlauskas, Jonas

AU - Peterlin Mašič, Lucíja

AU - Kunick, Conrad

AU - Labadie, Guillermo R.

AU - Flohé, Leopold

AU - Comini, Marcelo A.

PY - 2016/4/12

Y1 - 2016/4/12

N2 - Background: The search for novel chemical entities targeting essential and parasite-specific pathways is considered a priority for neglected diseases such as trypanosomiasis and leishmaniasis. The thiol-dependent redox metabolism of trypanosomatids relies on bis-glutathionylspermidine [trypanothione, T(SH)2], a low molecular mass cosubstrate absent in the host. In pathogenic trypanosomatids, a single enzyme, trypanothione synthetase (TryS), catalyzes trypanothione biosynthesis, which is indispensable for parasite survival. Thus, TryS qualifies as an attractive drug target candidate. Methodology/Principal Finding: A library composed of 144 compounds from 7 different families and several singletons was screened against TryS from three major pathogen species (Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum). The screening conditions were adjusted to the TryS´ kinetic parameters and intracellular concentration of substrates corresponding to each trypanosomatid species, and/or to avoid assay interference. The screening assay yielded suitable Z’ and signal to noise values (≥0.85 and ~3.5, respectively), and high intra-assay reproducibility. Several novel chemical scaffolds were identified as low μM and selective tri-tryp TryS inhibitors. Compounds displaying multi-TryS inhibition (N,N'-bis(3,4-substituted-benzyl) diamine derivatives) and an N5-substituted paullone (MOL2008) halted the proliferation of infective Trypanosoma brucei (EC50 in the nM range) and Leishmania infantum promastigotes (EC50 = 12 μM), respectively. A bis-benzyl diamine derivative and MOL2008 depleted intracellular trypanothione in treated parasites, which confirmed the on-target activity of these compounds. Conclusions/Significance: Novel molecular scaffolds with on-target mode of action were identified as hit candidates for TryS inhibition. Due to the remarkable species-specificity exhibited by tri-tryp TryS towards the compounds, future optimization and screening campaigns should aim at designing and detecting, respectively, more potent and broad-range TryS inhibitors.

AB - Background: The search for novel chemical entities targeting essential and parasite-specific pathways is considered a priority for neglected diseases such as trypanosomiasis and leishmaniasis. The thiol-dependent redox metabolism of trypanosomatids relies on bis-glutathionylspermidine [trypanothione, T(SH)2], a low molecular mass cosubstrate absent in the host. In pathogenic trypanosomatids, a single enzyme, trypanothione synthetase (TryS), catalyzes trypanothione biosynthesis, which is indispensable for parasite survival. Thus, TryS qualifies as an attractive drug target candidate. Methodology/Principal Finding: A library composed of 144 compounds from 7 different families and several singletons was screened against TryS from three major pathogen species (Trypanosoma brucei, Trypanosoma cruzi and Leishmania infantum). The screening conditions were adjusted to the TryS´ kinetic parameters and intracellular concentration of substrates corresponding to each trypanosomatid species, and/or to avoid assay interference. The screening assay yielded suitable Z’ and signal to noise values (≥0.85 and ~3.5, respectively), and high intra-assay reproducibility. Several novel chemical scaffolds were identified as low μM and selective tri-tryp TryS inhibitors. Compounds displaying multi-TryS inhibition (N,N'-bis(3,4-substituted-benzyl) diamine derivatives) and an N5-substituted paullone (MOL2008) halted the proliferation of infective Trypanosoma brucei (EC50 in the nM range) and Leishmania infantum promastigotes (EC50 = 12 μM), respectively. A bis-benzyl diamine derivative and MOL2008 depleted intracellular trypanothione in treated parasites, which confirmed the on-target activity of these compounds. Conclusions/Significance: Novel molecular scaffolds with on-target mode of action were identified as hit candidates for TryS inhibition. Due to the remarkable species-specificity exhibited by tri-tryp TryS towards the compounds, future optimization and screening campaigns should aim at designing and detecting, respectively, more potent and broad-range TryS inhibitors.

UR - http://www.scopus.com/inward/record.url?scp=84964883701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84964883701&partnerID=8YFLogxK

U2 - 10.1371/journal.pntd.0004617

DO - 10.1371/journal.pntd.0004617

M3 - Article

C2 - 27070550

AN - SCOPUS:84964883701

VL - 10

JO - PLoS Neglected Tropical Diseases

JF - PLoS Neglected Tropical Diseases

SN - 1935-2727

IS - 4

M1 - e0004617

ER -