Structure-guided approach to identify a novel class of anti-leishmaniasis diaryl sulfide compounds targeting the trypanothione metabolism

Colotti, Gianni; Saccoliti, Francesco; Gramiccia, Marina; Di Muccio, Trentina; Prakash, Jay; Yadav, Sunita; Dubey, Vikash Kumar; Vistoli, Giulio; Battista, Theo; Mocci, Stefano; Fiorillo, Annarita; Bibi, Aasia; Madia, Valentina Noemi; Messore, Antonella; Costi, Roberta; Di Santo, Roberto; Ilari, Andrea

doi:10.1007/s00726-019-02731-4

Original Article
Published: 29 April 2019

Structure-guided approach to identify a novel class of anti-leishmaniasis diaryl sulfide compounds targeting the trypanothione metabolism

Gianni Colotti¹,
Francesco Saccoliti³,
Marina Gramiccia⁴,
Trentina Di Muccio⁴,
Jay Prakash⁵,
Sunita Yadav⁶,
Vikash Kumar Dubey⁶,
Giulio Vistoli⁷,
Theo Battista²,
Stefano Mocci²,
Annarita Fiorillo²,
Aasia Bibi²,
Valentina Noemi Madia³,
Antonella Messore³,
Roberta Costi³,
Roberto Di Santo³ &
Andrea Ilari ORCID: orcid.org/0000-0002-7754-399X¹

Amino Acids volume 52, pages 247–259 (2020)Cite this article

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Abstract

Leishmania protozoans are the causative agent of leishmaniasis, a neglected tropical disease consisting of three major clinical forms: visceral leishmaniasis (VL), cutaneous leishmaniasis, and mucocutaneous leishmaniasis. VL is caused by Leishmania donovani in East Africa and the Indian subcontinent and by Leishmania infantum in Europe, North Africa, and Latin America, and causes an estimated 60,000 deaths per year. Trypanothione reductase (TR) is considered to be one of the best targets to find new drugs against leishmaniasis. This enzyme is fundamental for parasite survival in the human host since it reduces trypanothione, a molecule used by the tryparedoxin/tryparedoxin peroxidase system of Leishmania to neutralize the hydrogen peroxide produced by host macrophages during infection. Recently, we solved the X-ray structure of TR in complex with the diaryl sulfide compound RDS 777 (6-(sec-butoxy)-2-((3-chlorophenyl)thio)pyrimidin-4-amine), which impairs the parasite defense against the reactive oxygen species by inhibiting TR with high efficiency. The compound binds to the catalytic site and engages in hydrogen bonds the residues more involved in the catalysis, namely Glu466′, Cys57 and Cys52, thereby inhibiting the trypanothione binding. On the basis of the RDS 777–TR complex, we synthesized structurally related diaryl sulfide analogs as TR inhibitors able to compete for trypanothione binding to the enzyme and to kill the promastigote in the micromolar range. One of the most active among these compounds (RDS 562) was able to reduce the trypanothione concentration in cell of about 33% via TR inhibition. RDS 562 inhibits selectively Leishmania TR, while it does not inhibit the human homolog glutathione reductase.

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Abbreviations

FAD:: Flavin dinucleotide
GSSG:: Oxidized glutathione
hGR:: Human glutathione reductase
IR:: Infrared
NADPH:: Reduced nicotinamide adenine dinucleotide
TR:: Trypanothione reductase
TryS:: Trypanothione synthetase
TS₂ :: Trypanothione [N1,N8-bis(glutathionyl)spermidine]
T(SH)₂ :: Reduced trypanothione
TXN:: Tryparedoxin
TXNPx:: Tryparedoxin peroxidase I
VL:: Visceral leishmaniasis

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Acknowledgements

We gratefully acknowledge CNCCS CNR (National Collection of Chemical Compounds and Screening Center 2018) to AI and MIUR PRIN 20154JRJPP to GC which supported the experiments reported in these studies. This paper is dedicated to the memory of our friend and colleague Prof. Emilia Chiancone, passed away sadly in December 2018.

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Affiliations

Istituto di Biologia e Patologia Molecolari, Consiglio Nazionale delle Ricerche, IBPM-CNR, c/o Dip. Scienze Biochimiche Università Sapienza, P.le Aldo Moro 5, 00185, Rome, Italy
Gianni Colotti & Andrea Ilari
Dip. Scienze Biochimiche, Università Sapienza, P.le Aldo Moro 5, 00185, Rome, Italy
Theo Battista, Stefano Mocci, Annarita Fiorillo & Aasia Bibi
Dip. Chimica e Tecnologie del Farmaco, Istituto Pasteur-Fondazione Cenci Bolognetti, Università Sapienza, P.le Aldo Moro 5, 00185, Rome, Italy
Francesco Saccoliti, Valentina Noemi Madia, Antonella Messore, Roberta Costi & Roberto Di Santo
Dipartimento di Malattie Infettive, Istituto Superiore di Sanità Viale Regina Elena, 299, 00161, Rome, Italy
Marina Gramiccia & Trentina Di Muccio
Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam, 981039, India
Jay Prakash
School of Biochemical Engineering, Indian Institute of Technology (Banaras Hindu University) Varanasi, Varanasi, 221005, India
Sunita Yadav & Vikash Kumar Dubey
Dip. di Scienze Farmaceutiche, Università degli Studi di Milano, via Mangiagalli 25, 20133, Milan, Italy
Giulio Vistoli

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Gianni Colotti
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Francesco Saccoliti
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Marina Gramiccia
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Trentina Di Muccio
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Jay Prakash
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Sunita Yadav
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Vikash Kumar Dubey
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Giulio Vistoli
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Theo Battista
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Stefano Mocci
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Annarita Fiorillo
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Aasia Bibi
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Valentina Noemi Madia
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Antonella Messore
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Roberta Costi
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Roberto Di Santo
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Andrea Ilari
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Correspondence to Andrea Ilari.

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Colotti, G., Saccoliti, F., Gramiccia, M. et al. Structure-guided approach to identify a novel class of anti-leishmaniasis diaryl sulfide compounds targeting the trypanothione metabolism. Amino Acids 52, 247–259 (2020). https://doi.org/10.1007/s00726-019-02731-4

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Received: 18 January 2019
Accepted: 03 April 2019
Published: 29 April 2019
Issue Date: February 2020
DOI: https://doi.org/10.1007/s00726-019-02731-4

Keywords

Trypanothione metabolism
Trypanothione reductase
Structure-based drug design
Diaryl sulfide inhibitors