Investigation of the mycobacterial enzyme HsaD as a potential novel target for anti-tubercular agents using a fragment-based drug design approach

Ali Ryan, Elena Polycarpou, Nathan A. Lack, Dimitrios Evangelopoulos, Christian Sieg, Alice Halman, Sanjib Bhakta, Olga Eleftheriadou, Timothy D. McHugh, Sebastian Keany, Edward D. Lowe, Romain Ballet, Areej Abuhammad, William R. Jacobs, Alessio Ciulli, Edith Sim

Research output: Contribution to journalArticle

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Abstract

Background and Purpose: With the emergence of extensively drug-resistant tuberculosis, there is a need for new anti-tubercular drugs that work through novel mechanisms of action. The meta cleavage product hydrolase, HsaD, has been demonstrated to be critical for the survival of Mycobacterium tuberculosis in macrophages and is encoded in an operon involved in cholesterol catabolism, which is identical in M. tuberculosis and M. bovis BCG. Experimental Approach: We generated a mutant strain of M. bovis BCG with a deletion of hsaD and tested its growth on cholesterol. Using a fragment based approach, over 1000 compounds were screened by a combination of differential scanning fluorimetry, NMR spectroscopy and enzymatic assay with pure recombinant HsaD to identify potential inhibitors. We used enzymological and structural studies to investigate derivatives of the inhibitors identified and to test their effects on growth of M. bovis BCG and M. tuberculosis. Key Results: The hsaD deleted strain was unable to grow on cholesterol as sole carbon source but did grow on glucose. Of seven chemically distinct ‘hits’ from the library, two chemical classes of fragments were found to bind in the vicinity of the active site of HsaD by X-ray crystallography. The compounds also inhibited growth of M. tuberculosis on cholesterol. The most potent inhibitor of HsaD was also found to be the best inhibitor of mycobacterial growth on cholesterol-supplemented minimal medium. Conclusions and Implications: We propose that HsaD is a novel therapeutic target, which should be fully exploited in order to design and discover new anti-tubercular drugs. Linked Articles: This article is part of a themed section on Drug Metabolism and Antibiotic Resistance in Micro-organisms. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.14/issuetoc.

Original languageEnglish (US)
Pages (from-to)2209-2224
Number of pages16
JournalBritish Journal of Pharmacology
Volume174
Issue number14
DOIs
StatePublished - Jan 1 2017

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Drug Design
Cholesterol
Mycobacterium bovis
Enzymes
Tuberculosis
Growth
Extensively Drug-Resistant Tuberculosis
Small Molecule Libraries
Fluorometry
Growth Inhibitors
X Ray Crystallography
Enzyme Assays
Hydrolases
Operon
Microbial Drug Resistance
Mycobacterium tuberculosis
Drug Resistance
Pharmaceutical Preparations
Catalytic Domain
Magnetic Resonance Spectroscopy

ASJC Scopus subject areas

  • Pharmacology

Cite this

Investigation of the mycobacterial enzyme HsaD as a potential novel target for anti-tubercular agents using a fragment-based drug design approach. / Ryan, Ali; Polycarpou, Elena; Lack, Nathan A.; Evangelopoulos, Dimitrios; Sieg, Christian; Halman, Alice; Bhakta, Sanjib; Eleftheriadou, Olga; McHugh, Timothy D.; Keany, Sebastian; Lowe, Edward D.; Ballet, Romain; Abuhammad, Areej; Jacobs, William R.; Ciulli, Alessio; Sim, Edith.

In: British Journal of Pharmacology, Vol. 174, No. 14, 01.01.2017, p. 2209-2224.

Research output: Contribution to journalArticle

Ryan, A, Polycarpou, E, Lack, NA, Evangelopoulos, D, Sieg, C, Halman, A, Bhakta, S, Eleftheriadou, O, McHugh, TD, Keany, S, Lowe, ED, Ballet, R, Abuhammad, A, Jacobs, WR, Ciulli, A & Sim, E 2017, 'Investigation of the mycobacterial enzyme HsaD as a potential novel target for anti-tubercular agents using a fragment-based drug design approach', British Journal of Pharmacology, vol. 174, no. 14, pp. 2209-2224. https://doi.org/10.1111/bph.13810
Ryan, Ali ; Polycarpou, Elena ; Lack, Nathan A. ; Evangelopoulos, Dimitrios ; Sieg, Christian ; Halman, Alice ; Bhakta, Sanjib ; Eleftheriadou, Olga ; McHugh, Timothy D. ; Keany, Sebastian ; Lowe, Edward D. ; Ballet, Romain ; Abuhammad, Areej ; Jacobs, William R. ; Ciulli, Alessio ; Sim, Edith. / Investigation of the mycobacterial enzyme HsaD as a potential novel target for anti-tubercular agents using a fragment-based drug design approach. In: British Journal of Pharmacology. 2017 ; Vol. 174, No. 14. pp. 2209-2224.
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AU - Sieg, Christian

AU - Halman, Alice

AU - Bhakta, Sanjib

AU - Eleftheriadou, Olga

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AU - Keany, Sebastian

AU - Lowe, Edward D.

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