Identification of Leishmania major UDP-Sugar Pyrophosphorylase Inhibitors Using Biosensor-Based Small Molecule Fragment Library Screening

Ohm Prakash, Jana Führing, John Post, Sharon M. Shepherd, Thomas C. Eadsforth, David Gray, Roman Fedorov, Françoise H. Routier (Lead / Corresponding author)

Research output: Contribution to journalArticle

2 Citations (Scopus)
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Abstract

Leishmaniasis is a neglected disease that is caused by different species of the protozoan parasite Leishmania, and it currently affects 12 million people worldwide. The antileishmanial therapeutic arsenal remains very limited in number and efficacy, and there is no vaccine for this parasitic disease. One pathway that has been genetically validated as an antileishmanial drug target is the biosynthesis of uridine diphosphate-glucose (UDP-Glc), and its direct derivative UDP-galactose (UDP-Gal). De novo biosynthesis of these two nucleotide sugars is controlled by the specific UDP-glucose pyrophosphorylase (UGP). Leishmania parasites additionally express a UDP-sugar pyrophosphorylase (USP) responsible for monosaccharides salvage that is able to generate both UDP-Gal and UDP-Glc. The inactivation of the two parasite pyrophosphorylases UGP and USP, results in parasite death. The present study reports on the identification of structurally diverse scaffolds for the development of USP inhibitors by fragment library screening. Based on this screening, we selected a small set of commercially available compounds, and identified molecules that inhibit both Leishmania major USP and UGP, with a half-maximal inhibitory concentration in the 100 µM range. The inhibitors were predicted to bind at allosteric regulation sites, which were validated by mutagenesis studies. This study sets the stage for the development of potent USP inhibitors.

Original languageEnglish
Article number996
Pages (from-to)1-16
Number of pages16
JournalMolecules and Cells
Volume24
Issue number5
DOIs
Publication statusPublished - 12 Mar 2019

Fingerprint

Uridine Diphosphate Sugars
Small Molecule Libraries
Leishmania major
Biosensing Techniques
sugars
bioinstrumentation
UTP-Glucose-1-Phosphate Uridylyltransferase
Biosensors
inhibitors
parasites
Screening
glucose
screening
fragments
Parasites
Molecules
Uridine Diphosphate Glucose
Uridine Diphosphate
Leishmania
Biosynthesis

Keywords

  • Hematopoietic stem cells
  • NK cell therapy
  • NK receptor repertoire
  • Natural killer cell differentiation

Cite this

Prakash, Ohm ; Führing, Jana ; Post, John ; Shepherd, Sharon M. ; Eadsforth, Thomas C. ; Gray, David ; Fedorov, Roman ; Routier, Françoise H. / Identification of Leishmania major UDP-Sugar Pyrophosphorylase Inhibitors Using Biosensor-Based Small Molecule Fragment Library Screening. In: Molecules and Cells. 2019 ; Vol. 24, No. 5. pp. 1-16.
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Identification of Leishmania major UDP-Sugar Pyrophosphorylase Inhibitors Using Biosensor-Based Small Molecule Fragment Library Screening. / Prakash, Ohm; Führing, Jana; Post, John; Shepherd, Sharon M.; Eadsforth, Thomas C.; Gray, David; Fedorov, Roman; Routier, Françoise H. (Lead / Corresponding author).

In: Molecules and Cells, Vol. 24, No. 5, 996, 12.03.2019, p. 1-16.

Research output: Contribution to journalArticle

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AU - Routier, Françoise H.

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AB - Leishmaniasis is a neglected disease that is caused by different species of the protozoan parasite Leishmania, and it currently affects 12 million people worldwide. The antileishmanial therapeutic arsenal remains very limited in number and efficacy, and there is no vaccine for this parasitic disease. One pathway that has been genetically validated as an antileishmanial drug target is the biosynthesis of uridine diphosphate-glucose (UDP-Glc), and its direct derivative UDP-galactose (UDP-Gal). De novo biosynthesis of these two nucleotide sugars is controlled by the specific UDP-glucose pyrophosphorylase (UGP). Leishmania parasites additionally express a UDP-sugar pyrophosphorylase (USP) responsible for monosaccharides salvage that is able to generate both UDP-Gal and UDP-Glc. The inactivation of the two parasite pyrophosphorylases UGP and USP, results in parasite death. The present study reports on the identification of structurally diverse scaffolds for the development of USP inhibitors by fragment library screening. Based on this screening, we selected a small set of commercially available compounds, and identified molecules that inhibit both Leishmania major USP and UGP, with a half-maximal inhibitory concentration in the 100 µM range. The inhibitors were predicted to bind at allosteric regulation sites, which were validated by mutagenesis studies. This study sets the stage for the development of potent USP inhibitors.

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