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Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani

[Image: see text] Visceral leishmaniasis (VL), caused by the protozoan parasites Leishmania donovani and L. infantum, is responsible for ∼30 000 deaths annually. Available treatments are inadequate, and there is a pressing need for new therapeutics. N-Myristoyltransferase (NMT) remains one of the fe...

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Autores principales: Corpas-Lopez, Victoriano, Moniz, Sonia, Thomas, Michael, Wall, Richard J., Torrie, Leah S., Zander-Dinse, Dorothea, Tinti, Michele, Brand, Stephen, Stojanovski, Laste, Manthri, Sujatha, Hallyburton, Irene, Zuccotto, Fabio, Wyatt, Paul G., De Rycker, Manu, Horn, David, Ferguson, Michael A. J., Clos, Joachim, Read, Kevin D., Fairlamb, Alan H., Gilbert, Ian H., Wyllie, Susan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2018
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332449/
https://www.ncbi.nlm.nih.gov/pubmed/30380837
http://dx.doi.org/10.1021/acsinfecdis.8b00226
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author Corpas-Lopez, Victoriano
Moniz, Sonia
Thomas, Michael
Wall, Richard J.
Torrie, Leah S.
Zander-Dinse, Dorothea
Tinti, Michele
Brand, Stephen
Stojanovski, Laste
Manthri, Sujatha
Hallyburton, Irene
Zuccotto, Fabio
Wyatt, Paul G.
De Rycker, Manu
Horn, David
Ferguson, Michael A. J.
Clos, Joachim
Read, Kevin D.
Fairlamb, Alan H.
Gilbert, Ian H.
Wyllie, Susan
author_facet Corpas-Lopez, Victoriano
Moniz, Sonia
Thomas, Michael
Wall, Richard J.
Torrie, Leah S.
Zander-Dinse, Dorothea
Tinti, Michele
Brand, Stephen
Stojanovski, Laste
Manthri, Sujatha
Hallyburton, Irene
Zuccotto, Fabio
Wyatt, Paul G.
De Rycker, Manu
Horn, David
Ferguson, Michael A. J.
Clos, Joachim
Read, Kevin D.
Fairlamb, Alan H.
Gilbert, Ian H.
Wyllie, Susan
author_sort Corpas-Lopez, Victoriano
collection PubMed
description [Image: see text] Visceral leishmaniasis (VL), caused by the protozoan parasites Leishmania donovani and L. infantum, is responsible for ∼30 000 deaths annually. Available treatments are inadequate, and there is a pressing need for new therapeutics. N-Myristoyltransferase (NMT) remains one of the few genetically validated drug targets in these parasites. Here, we sought to pharmacologically validate this enzyme in Leishmania. A focused set of 1600 pyrazolyl sulfonamide compounds was screened against L. major NMT in a robust high-throughput biochemical assay. Several potent inhibitors were identified with marginal selectivity over the human enzyme. There was little correlation between the enzyme potency of these inhibitors and their cellular activity against L. donovani axenic amastigotes, and this discrepancy could be due to poor cellular uptake due to the basicity of these compounds. Thus, a series of analogues were synthesized with less basic centers. Although most of these compounds continued to suffer from relatively poor antileishmanial activity, our most potent inhibitor of LmNMT (DDD100097, K(i) of 0.34 nM) showed modest activity against L. donovani intracellular amastigotes (EC(50) of 2.4 μM) and maintained a modest therapeutic window over the human enzyme. Two unbiased approaches, namely, screening against our cosmid-based overexpression library and thermal proteome profiling (TPP), confirm that DDD100097 (compound 2) acts on-target within parasites. Oral dosing with compound 2 resulted in a 52% reduction in parasite burden in our mouse model of VL. Thus, NMT is now a pharmacologically validated target in Leishmania. The challenge in finding drug candidates remains to identify alternative strategies to address the drop-off in activity between enzyme inhibition and in vitro activity while maintaining sufficient selectivity over the human enzyme, both issues that continue to plague studies in this area.
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spelling pubmed-63324492019-01-17 Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani Corpas-Lopez, Victoriano Moniz, Sonia Thomas, Michael Wall, Richard J. Torrie, Leah S. Zander-Dinse, Dorothea Tinti, Michele Brand, Stephen Stojanovski, Laste Manthri, Sujatha Hallyburton, Irene Zuccotto, Fabio Wyatt, Paul G. De Rycker, Manu Horn, David Ferguson, Michael A. J. Clos, Joachim Read, Kevin D. Fairlamb, Alan H. Gilbert, Ian H. Wyllie, Susan ACS Infect Dis [Image: see text] Visceral leishmaniasis (VL), caused by the protozoan parasites Leishmania donovani and L. infantum, is responsible for ∼30 000 deaths annually. Available treatments are inadequate, and there is a pressing need for new therapeutics. N-Myristoyltransferase (NMT) remains one of the few genetically validated drug targets in these parasites. Here, we sought to pharmacologically validate this enzyme in Leishmania. A focused set of 1600 pyrazolyl sulfonamide compounds was screened against L. major NMT in a robust high-throughput biochemical assay. Several potent inhibitors were identified with marginal selectivity over the human enzyme. There was little correlation between the enzyme potency of these inhibitors and their cellular activity against L. donovani axenic amastigotes, and this discrepancy could be due to poor cellular uptake due to the basicity of these compounds. Thus, a series of analogues were synthesized with less basic centers. Although most of these compounds continued to suffer from relatively poor antileishmanial activity, our most potent inhibitor of LmNMT (DDD100097, K(i) of 0.34 nM) showed modest activity against L. donovani intracellular amastigotes (EC(50) of 2.4 μM) and maintained a modest therapeutic window over the human enzyme. Two unbiased approaches, namely, screening against our cosmid-based overexpression library and thermal proteome profiling (TPP), confirm that DDD100097 (compound 2) acts on-target within parasites. Oral dosing with compound 2 resulted in a 52% reduction in parasite burden in our mouse model of VL. Thus, NMT is now a pharmacologically validated target in Leishmania. The challenge in finding drug candidates remains to identify alternative strategies to address the drop-off in activity between enzyme inhibition and in vitro activity while maintaining sufficient selectivity over the human enzyme, both issues that continue to plague studies in this area. American Chemical Society 2018-11-01 2019-01-11 /pmc/articles/PMC6332449/ /pubmed/30380837 http://dx.doi.org/10.1021/acsinfecdis.8b00226 Text en Copyright © 2018 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Corpas-Lopez, Victoriano
Moniz, Sonia
Thomas, Michael
Wall, Richard J.
Torrie, Leah S.
Zander-Dinse, Dorothea
Tinti, Michele
Brand, Stephen
Stojanovski, Laste
Manthri, Sujatha
Hallyburton, Irene
Zuccotto, Fabio
Wyatt, Paul G.
De Rycker, Manu
Horn, David
Ferguson, Michael A. J.
Clos, Joachim
Read, Kevin D.
Fairlamb, Alan H.
Gilbert, Ian H.
Wyllie, Susan
Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title_full Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title_fullStr Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title_full_unstemmed Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title_short Pharmacological Validation of N-Myristoyltransferase as a Drug Target in Leishmania donovani
title_sort pharmacological validation of n-myristoyltransferase as a drug target in leishmania donovani
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6332449/
https://www.ncbi.nlm.nih.gov/pubmed/30380837
http://dx.doi.org/10.1021/acsinfecdis.8b00226
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