A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells

A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells

ABSTRACT Active tuberculosis (TB) and latent Mycobacterium tuberculosis infection both require lengthy treatments to achieve durable cures. This problem has partly been attributable to the existence of nonreplicating M. tuberculosis “persisters” that are difficult to kill using conventional anti-TB...

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Autores principales: Paridhi Sukheja, Pradeep Kumar, Nisha Mittal, Shao-Gang Li, Eric Singleton, Riccardo Russo, Alexander L. Perryman, Riju Shrestha, Divya Awasthi, Seema Husain, Patricia Soteropoulos, Roman Brukh, Nancy Connell, Joel S. Freundlich, David Alland
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Publicado: American Society for Microbiology 2017
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Microbiology
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spelling oai:doaj.org-article:4063dfd78f48410f8cdd22b4c8e25ff72021-11-15T15:51:06ZA Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells10.1128/mBio.02022-162150-7511https://doaj.org/article/4063dfd78f48410f8cdd22b4c8e25ff72017-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02022-16https://doaj.org/toc/2150-7511ABSTRACT Active tuberculosis (TB) and latent Mycobacterium tuberculosis infection both require lengthy treatments to achieve durable cures. This problem has partly been attributable to the existence of nonreplicating M. tuberculosis “persisters” that are difficult to kill using conventional anti-TB treatments. Compounds that target the respiratory pathway have the potential to kill both replicating and persistent M. tuberculosis and shorten TB treatment, as this pathway is essential in both metabolic states. We developed a novel respiratory pathway-specific whole-cell screen to identify new respiration inhibitors. This screen identified the biphenyl amide GSK1733953A (DG70) as a likely respiration inhibitor. DG70 inhibited both clinical drug-susceptible and drug-resistant M. tuberculosis strains. Whole-genome sequencing of DG70-resistant colonies identified mutations in menG (rv0558), which is responsible for the final step in menaquinone biosynthesis and required for respiration. Overexpression of menG from wild-type and DG70-resistant isolates increased the DG70 MIC by 4× and 8× to 30×, respectively. Radiolabeling and high-resolution mass spectrometry studies confirmed that DG70 inhibited the final step in menaquinone biosynthesis. DG70 also inhibited oxygen utilization and ATP biosynthesis, which was reversed by external menaquinone supplementation. DG70 was bactericidal in actively replicating cultures and in a nutritionally deprived persistence model. DG70 was synergistic with the first-line TB drugs isoniazid, rifampin, and the respiratory inhibitor bedaquiline. The combination of DG70 and isoniazid completely sterilized cultures in the persistence model by day 10. These results suggest that MenG is a good therapeutic target and that compounds targeting MenG along with standard TB therapy have the potential to shorten TB treatment duration. IMPORTANCE This study shows that MenG, which is responsible for the last enzymatic step in menaquinone biosynthesis, may be a good drug target for improving TB treatments. We describe the first small-molecule inhibitor (DG70) of Mycobacterium tuberculosis MenG and show that DG70 has characteristics that are highly desirable for a new antitubercular agent, including bactericidality against both actively growing and nonreplicating mycobacteria and synergy with several first-line drugs that are currently used to treat TB.Paridhi SukhejaPradeep KumarNisha MittalShao-Gang LiEric SingletonRiccardo RussoAlexander L. PerrymanRiju ShresthaDivya AwasthiSeema HusainPatricia SoteropoulosRoman BrukhNancy ConnellJoel S. FreundlichDavid AllandAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 8, Iss 1 (2017)
institution DOAJ
collection DOAJ
language EN
topic Microbiology
QR1-502
spellingShingle Microbiology
QR1-502
Paridhi Sukheja
Pradeep Kumar
Nisha Mittal
Shao-Gang Li
Eric Singleton
Riccardo Russo
Alexander L. Perryman
Riju Shrestha
Divya Awasthi
Seema Husain
Patricia Soteropoulos
Roman Brukh
Nancy Connell
Joel S. Freundlich
David Alland
A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
description ABSTRACT Active tuberculosis (TB) and latent Mycobacterium tuberculosis infection both require lengthy treatments to achieve durable cures. This problem has partly been attributable to the existence of nonreplicating M. tuberculosis “persisters” that are difficult to kill using conventional anti-TB treatments. Compounds that target the respiratory pathway have the potential to kill both replicating and persistent M. tuberculosis and shorten TB treatment, as this pathway is essential in both metabolic states. We developed a novel respiratory pathway-specific whole-cell screen to identify new respiration inhibitors. This screen identified the biphenyl amide GSK1733953A (DG70) as a likely respiration inhibitor. DG70 inhibited both clinical drug-susceptible and drug-resistant M. tuberculosis strains. Whole-genome sequencing of DG70-resistant colonies identified mutations in menG (rv0558), which is responsible for the final step in menaquinone biosynthesis and required for respiration. Overexpression of menG from wild-type and DG70-resistant isolates increased the DG70 MIC by 4× and 8× to 30×, respectively. Radiolabeling and high-resolution mass spectrometry studies confirmed that DG70 inhibited the final step in menaquinone biosynthesis. DG70 also inhibited oxygen utilization and ATP biosynthesis, which was reversed by external menaquinone supplementation. DG70 was bactericidal in actively replicating cultures and in a nutritionally deprived persistence model. DG70 was synergistic with the first-line TB drugs isoniazid, rifampin, and the respiratory inhibitor bedaquiline. The combination of DG70 and isoniazid completely sterilized cultures in the persistence model by day 10. These results suggest that MenG is a good therapeutic target and that compounds targeting MenG along with standard TB therapy have the potential to shorten TB treatment duration. IMPORTANCE This study shows that MenG, which is responsible for the last enzymatic step in menaquinone biosynthesis, may be a good drug target for improving TB treatments. We describe the first small-molecule inhibitor (DG70) of Mycobacterium tuberculosis MenG and show that DG70 has characteristics that are highly desirable for a new antitubercular agent, including bactericidality against both actively growing and nonreplicating mycobacteria and synergy with several first-line drugs that are currently used to treat TB.
format article
author Paridhi Sukheja
Pradeep Kumar
Nisha Mittal
Shao-Gang Li
Eric Singleton
Riccardo Russo
Alexander L. Perryman
Riju Shrestha
Divya Awasthi
Seema Husain
Patricia Soteropoulos
Roman Brukh
Nancy Connell
Joel S. Freundlich
David Alland
author_facet Paridhi Sukheja
Pradeep Kumar
Nisha Mittal
Shao-Gang Li
Eric Singleton
Riccardo Russo
Alexander L. Perryman
Riju Shrestha
Divya Awasthi
Seema Husain
Patricia Soteropoulos
Roman Brukh
Nancy Connell
Joel S. Freundlich
David Alland
author_sort Paridhi Sukheja
title A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
title_short A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
title_full A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
title_fullStr A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
title_full_unstemmed A Novel Small-Molecule Inhibitor of the <italic toggle="yes">Mycobacterium tuberculosis</italic> Demethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells
title_sort novel small-molecule inhibitor of the <italic toggle="yes">mycobacterium tuberculosis</italic> demethylmenaquinone methyltransferase meng is bactericidal to both growing and nutritionally deprived persister cells
publisher American Society for Microbiology
publishDate 2017
url https://doaj.org/article/4063dfd78f48410f8cdd22b4c8e25ff7
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