Chemical Validation of Methionyl tRNA Synthetase (MetRS) as a Druggable Target in Leishmania donovani

Leah Torrie, Stephen Brand, David Robinson, Eun Jung Ko, Laste Stojanovski, Frederick R. C. Simeons, Susan Wyllie, John Thomas, Lucy Ellis, Maria Osuna-Cabello, Rafiu Epemolu, Andrea Nuehs, Jennifer Riley, Lorna MacLean, Sujatha Manthri, Kevin Read, Ian Gilbert (Lead / Corresponding author), Alan Fairlamb, Manu De Rycker (Lead / Corresponding author)

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Abstract

Methionyl-tRNA synthetase (MetRS) has been chemically validated as a drug target in the kinetoplastid parasite Trypanosoma brucei. In the present study we investigate the validity of this target in the related trypanosomatid Leishmania donovani. Following development of a robust highthroughput compatible biochemical assay, a compound screen identified DDD806905 as a highly potent inhibitor of LdMetRS (Ki 18 nM). Crystallography revealed this compound binds to the methionine pocket of MetRS with enzymatic studies confirming DDD806905 displays competitive inhibition with respect to methionine and mixed inhibition with respect to ATP binding. DDD806905 showed activity, albeit with different levels of potency, in various Leishmania cell-based viability assays, with on-target activity observed in both Leishmania promastigote cell assays and a Leishmania tarentolae in vitro translation assay. Unfortunately this compound failed to show efficacy in an animal model of leishmaniasis. We investigated the potential causes for the discrepancies in activity observed in different Leishmania cell assays and the lack of efficacy in the animal model and found that high protein binding as well as sequestration of this dibasic compound into acidic compartments may play a role. Despite medicinal chemistry efforts to address the dibasic nature of DDD806905 and analogues, no progress could be achieved with the current chemical series. Although DDD806905 is not a developable anti-leishmanial compound, MetRS remains an attractive anti-leishmanial drug target.
Original languageEnglish
Pages (from-to)718-727
Number of pages10
JournalACS Infectious Diseases
Volume3
Issue number10
Early online date2 Oct 2017
DOIs
Publication statusPublished - Oct 2017

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Methionine-tRNA Ligase
Leishmania donovani
Leishmania
Methionine
Animal Models
Trypanosoma brucei brucei
Crystallography
Leishmaniasis
Pharmaceutical Chemistry
Protein Binding
Pharmaceutical Preparations
Cell Survival
Parasites
Adenosine Triphosphate

Keywords

  • Drug Discovery
  • Kinetoplastids
  • Translation
  • tRNA synthetase
  • Parasite

Cite this

@article{efe228b9806c4c778cb5c9e519f8a5d4,
title = "Chemical Validation of Methionyl tRNA Synthetase (MetRS) as a Druggable Target in Leishmania donovani",
abstract = "Methionyl-tRNA synthetase (MetRS) has been chemically validated as a drug target in the kinetoplastid parasite Trypanosoma brucei. In the present study we investigate the validity of this target in the related trypanosomatid Leishmania donovani. Following development of a robust highthroughput compatible biochemical assay, a compound screen identified DDD806905 as a highly potent inhibitor of LdMetRS (Ki 18 nM). Crystallography revealed this compound binds to the methionine pocket of MetRS with enzymatic studies confirming DDD806905 displays competitive inhibition with respect to methionine and mixed inhibition with respect to ATP binding. DDD806905 showed activity, albeit with different levels of potency, in various Leishmania cell-based viability assays, with on-target activity observed in both Leishmania promastigote cell assays and a Leishmania tarentolae in vitro translation assay. Unfortunately this compound failed to show efficacy in an animal model of leishmaniasis. We investigated the potential causes for the discrepancies in activity observed in different Leishmania cell assays and the lack of efficacy in the animal model and found that high protein binding as well as sequestration of this dibasic compound into acidic compartments may play a role. Despite medicinal chemistry efforts to address the dibasic nature of DDD806905 and analogues, no progress could be achieved with the current chemical series. Although DDD806905 is not a developable anti-leishmanial compound, MetRS remains an attractive anti-leishmanial drug target.",
keywords = "Drug Discovery, Kinetoplastids, Translation, tRNA synthetase, Parasite",
author = "Leah Torrie and Stephen Brand and David Robinson and Ko, {Eun Jung} and Laste Stojanovski and Simeons, {Frederick R. C.} and Susan Wyllie and John Thomas and Lucy Ellis and Maria Osuna-Cabello and Rafiu Epemolu and Andrea Nuehs and Jennifer Riley and Lorna MacLean and Sujatha Manthri and Kevin Read and Ian Gilbert and Alan Fairlamb and {De Rycker}, Manu",
note = "We would like to acknowledge the Wellcome Trust for funding (grants 092340, 105021, 100476). We would like to acknowledge GSK for provision of the TCAMS (Tres Cantos Anti-Malarial Set) for screening against the kinetoplastids and some additional bacterial MetRS inhibitors. We also acknowledge the support of the Protein Production Team in the Division of Biological Chemistry and Drug Discovery (Dundee), the Drug Discovery Unit compound management, and Drug Discovery Unit data management teams. The authors would like to thank Diamond Light Source for beamtime (proposal mx8268), and the staff of beamlines I02 and I04 for assistance with data collection",
year = "2017",
month = "10",
doi = "10.1021/acsinfecdis.7b00047",
language = "English",
volume = "3",
pages = "718--727",
journal = "ACS Infectious Diseases",
issn = "2373-8227",
publisher = "American Chemical Society",
number = "10",

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TY - JOUR

T1 - Chemical Validation of Methionyl tRNA Synthetase (MetRS) as a Druggable Target in Leishmania donovani

AU - Torrie, Leah

AU - Brand, Stephen

AU - Robinson, David

AU - Ko, Eun Jung

AU - Stojanovski, Laste

AU - Simeons, Frederick R. C.

AU - Wyllie, Susan

AU - Thomas, John

AU - Ellis, Lucy

AU - Osuna-Cabello, Maria

AU - Epemolu, Rafiu

AU - Nuehs, Andrea

AU - Riley, Jennifer

AU - MacLean, Lorna

AU - Manthri, Sujatha

AU - Read, Kevin

AU - Gilbert, Ian

AU - Fairlamb, Alan

AU - De Rycker, Manu

N1 - We would like to acknowledge the Wellcome Trust for funding (grants 092340, 105021, 100476). We would like to acknowledge GSK for provision of the TCAMS (Tres Cantos Anti-Malarial Set) for screening against the kinetoplastids and some additional bacterial MetRS inhibitors. We also acknowledge the support of the Protein Production Team in the Division of Biological Chemistry and Drug Discovery (Dundee), the Drug Discovery Unit compound management, and Drug Discovery Unit data management teams. The authors would like to thank Diamond Light Source for beamtime (proposal mx8268), and the staff of beamlines I02 and I04 for assistance with data collection

PY - 2017/10

Y1 - 2017/10

N2 - Methionyl-tRNA synthetase (MetRS) has been chemically validated as a drug target in the kinetoplastid parasite Trypanosoma brucei. In the present study we investigate the validity of this target in the related trypanosomatid Leishmania donovani. Following development of a robust highthroughput compatible biochemical assay, a compound screen identified DDD806905 as a highly potent inhibitor of LdMetRS (Ki 18 nM). Crystallography revealed this compound binds to the methionine pocket of MetRS with enzymatic studies confirming DDD806905 displays competitive inhibition with respect to methionine and mixed inhibition with respect to ATP binding. DDD806905 showed activity, albeit with different levels of potency, in various Leishmania cell-based viability assays, with on-target activity observed in both Leishmania promastigote cell assays and a Leishmania tarentolae in vitro translation assay. Unfortunately this compound failed to show efficacy in an animal model of leishmaniasis. We investigated the potential causes for the discrepancies in activity observed in different Leishmania cell assays and the lack of efficacy in the animal model and found that high protein binding as well as sequestration of this dibasic compound into acidic compartments may play a role. Despite medicinal chemistry efforts to address the dibasic nature of DDD806905 and analogues, no progress could be achieved with the current chemical series. Although DDD806905 is not a developable anti-leishmanial compound, MetRS remains an attractive anti-leishmanial drug target.

AB - Methionyl-tRNA synthetase (MetRS) has been chemically validated as a drug target in the kinetoplastid parasite Trypanosoma brucei. In the present study we investigate the validity of this target in the related trypanosomatid Leishmania donovani. Following development of a robust highthroughput compatible biochemical assay, a compound screen identified DDD806905 as a highly potent inhibitor of LdMetRS (Ki 18 nM). Crystallography revealed this compound binds to the methionine pocket of MetRS with enzymatic studies confirming DDD806905 displays competitive inhibition with respect to methionine and mixed inhibition with respect to ATP binding. DDD806905 showed activity, albeit with different levels of potency, in various Leishmania cell-based viability assays, with on-target activity observed in both Leishmania promastigote cell assays and a Leishmania tarentolae in vitro translation assay. Unfortunately this compound failed to show efficacy in an animal model of leishmaniasis. We investigated the potential causes for the discrepancies in activity observed in different Leishmania cell assays and the lack of efficacy in the animal model and found that high protein binding as well as sequestration of this dibasic compound into acidic compartments may play a role. Despite medicinal chemistry efforts to address the dibasic nature of DDD806905 and analogues, no progress could be achieved with the current chemical series. Although DDD806905 is not a developable anti-leishmanial compound, MetRS remains an attractive anti-leishmanial drug target.

KW - Drug Discovery

KW - Kinetoplastids

KW - Translation

KW - tRNA synthetase

KW - Parasite

U2 - 10.1021/acsinfecdis.7b00047

DO - 10.1021/acsinfecdis.7b00047

M3 - Article

C2 - 28967262

VL - 3

SP - 718

EP - 727

JO - ACS Infectious Diseases

JF - ACS Infectious Diseases

SN - 2373-8227

IS - 10

ER -