TY - JOUR
T1 - Antituberculosis thiophenes define a requirement for Pks13 in mycolic acid biosynthesis
AU - Wilson, Regina
AU - Kumar, Pradeep
AU - Parashar, Vijay
AU - Vilchèze, Catherine
AU - Veyron-Churlet, Romain
AU - Freundlich, Joel S.
AU - Barnes, S. Whitney
AU - Walker, John R.
AU - Szymonifka, Michael J.
AU - Marchiano, Emily
AU - Shenai, Shubhada
AU - Colangeli, Roberto
AU - Jacobs, William R.
AU - Neiditch, Matthew B.
AU - Kremer, Laurent
AU - Alland, David
N1 - Funding Information:
This work was supported in part by US National Institutes of Health (NIH) grant R01 AI080653 to D.A., a United Negro College Fund–Merck Postdoctoral Science Research Fellowship to R.W., a grant from the University of Medicine and Dentistry of New Jersey (UMDNJ) foundation to R.C. and NIH grant R01 AI081736 to M.B.N. W.R.J. acknowledges generous support from the NIH Centers for AIDS Research grant AI-051519 at the Albert Einstein College of Medicine and by NIH grant AI26170. The compounds initially screened in this work were supplied as part of NIH grants AI-95364 and AI-15449. We thank R.C. Goldman (formally Department of Health and Human Services, NIH, and Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases (NIAID), currently RCG Consulting) for his support obtaining Molecular Libraries Probe Centers Network (MLPCN) compounds for the piniBAC screen, R. Reynolds (Southern Research Institute) for his advice on the initial analysis of the MLPCN library and C.E. Barry, III and H.I.M. Boshoff (Tuberculosis Research Section, Laboratory of Clinical Infectious Diseases, NIAID, NIH) for their kind gift of TDM and TMM standards. The MS data were obtained from an Orbitrap instrument funded in part by NIH grant NS046593 for the support of the UMDNJ Neuroproteomics Core Facility.
PY - 2013/8
Y1 - 2013/8
N2 - We report a new class of thiophene (TP) compounds that kill Mycobacterium tuberculosis by the previously uncharacterized mechanism of Pks13 inhibition. An F79S mutation near the catalytic Ser55 site in Pks13 conferred TP resistance in M. tuberculosis. Overexpression of wild-type Pks13 resulted in TP resistance, and overexpression of the Pks13 F79S mutant conferred high resistance. In vitro, TP inhibited fatty acyl-AMP loading onto Pks13. TP inhibited mycolic acid biosynthesis in wild-type M. tuberculosis, but it did so to a much lesser extent in TP-resistant M. tuberculosis. TP treatment was bactericidal and equivalent to treatment with the first-line drug isoniazid, but it was less likely to permit emergent resistance. Combined isoniazid and TP treatment resulted in sterilizing activity. Computational docking identified a possible TP-binding groove within the Pks13 acyl carrier protein domain. This study confirms that M. tuberculosis Pks13 is required for mycolic acid biosynthesis, validates it as a druggable target and demonstrates the therapeutic potential of simultaneously inhibiting multiple targets in the same biosynthetic pathway.
AB - We report a new class of thiophene (TP) compounds that kill Mycobacterium tuberculosis by the previously uncharacterized mechanism of Pks13 inhibition. An F79S mutation near the catalytic Ser55 site in Pks13 conferred TP resistance in M. tuberculosis. Overexpression of wild-type Pks13 resulted in TP resistance, and overexpression of the Pks13 F79S mutant conferred high resistance. In vitro, TP inhibited fatty acyl-AMP loading onto Pks13. TP inhibited mycolic acid biosynthesis in wild-type M. tuberculosis, but it did so to a much lesser extent in TP-resistant M. tuberculosis. TP treatment was bactericidal and equivalent to treatment with the first-line drug isoniazid, but it was less likely to permit emergent resistance. Combined isoniazid and TP treatment resulted in sterilizing activity. Computational docking identified a possible TP-binding groove within the Pks13 acyl carrier protein domain. This study confirms that M. tuberculosis Pks13 is required for mycolic acid biosynthesis, validates it as a druggable target and demonstrates the therapeutic potential of simultaneously inhibiting multiple targets in the same biosynthetic pathway.
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U2 - 10.1038/nchembio.1277
DO - 10.1038/nchembio.1277
M3 - Article
C2 - 23770708
AN - SCOPUS:84880924467
SN - 1552-4450
VL - 9
SP - 499
EP - 506
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 8
ER -
