title: A Novel Small-Molecule Inhibitor of theMycobacterium tuberculosisDemethylmenaquinone Methyltransferase MenG Is Bactericidal to Both Growing and Nutritionally Deprived Persister Cells.
abstract: Active tuberculosis (TB) and latentMycobacterium tuberculosisinfection both require lengthy treatments to achieve durable cures. This problem has partly been attributable to the existence of nonreplicatingM. 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 persistentM. tuberculosisand 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-resistantM. tuberculosisstrains. Whole-genome sequencing of DG70-resistant colonies identified mutations inmenG(rv0558), which is responsible for the final step in menaquinone biosynthesis and required for respiration. Overexpression ofmenGfrom 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.IMPORTANCEThis 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) ofMycobacterium tuberculosisMenG 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.
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Paridhi
authors.initials: P
authors.lastname: Sukheja
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Nisha
authors.initials: N
authors.lastname: Mittal
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Shao-Gang
authors.initials: SG
authors.lastname: Li
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Singleton
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Russo
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Alexander L
authors.initials: AL
authors.lastname: Perryman
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Riju
authors.initials: R
authors.lastname: Shrestha
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Divya
authors.initials: D
authors.lastname: Awasthi
authors.affiliation: Genomics Center, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.|Department of Microbiology, Biochemistry and Molecular Genetics, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Seema
authors.initials: S
authors.lastname: Husain
authors.affiliation: Genomics Center, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.|Department of Microbiology, Biochemistry and Molecular Genetics, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Soteropoulos
authors.affiliation: Department of Chemistry-Rutgers University-Newark College of Arts and Sciences, Newark, New Jersey, USA.
authors.forename: Roman
authors.initials: R
authors.lastname: Brukh
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Nancy
authors.initials: N
authors.lastname: Connell
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.|Department of Pharmacology, Physiology, and Neuroscience, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA.
authors.forename: Joel S
authors.initials: JS
authors.lastname: Freundlich
authors.affiliation: Division of Infectious Disease, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, Rutgers University-New Jersey Medical School, Newark, New Jersey, USA david.alland@rutgers.edu.
authors.forename: David
authors.initials: D
authors.lastname: Alland
mesh_terms: D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000995:Antitubercular Agents / Q000494:pharmacology*; D001713:Biphenyl Compounds / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D055808:Drug Discovery; D024881:Drug Resistance, Bacterial; D006801:Humans; D008780:Methyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology / Q000254:growth & development*; D054852:Small Molecule Libraries / Q000032:analysis; D024482:Vitamin K 2 / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000995:Antitubercular Agents; D001713:Biphenyl Compounds; D054852:Small Molecule Libraries; D024482:Vitamin K 2; C013917:demethylmenaquinone; D000255:Adenosine Triphosphate; D008780:Methyltransferases