title: Identification of arylamine N-acetyltransferase inhibitors as an approach towards novel anti-tuberculars.
abstract: New anti-tubercular drugs and drug targets are urgently needed to reduce the time for treatment and also to identify agents that will be effective against Mycobacterium tuberculosis persisting intracellularly. Mycobacteria have a unique cell wall. Deletion of the gene for arylamine N-acetyltransferase (NAT) decreases mycobacterial cell wall lipids, particularly the distinctive mycolates, and also increases antibiotic susceptibility and killing within macrophage of Mycobacterium bovis BCG. The nat gene and its associated gene cluster are almost identical in sequence in M. bovis BCG and M. tuberculosis. The gene cluster is essential for intracellular survival of mycobacteria. We have therefore used pure NAT protein for high-throughput screening to identify several classes of small molecules that inhibit NAT activity. Here, we characterize one class of such molecules-triazoles-in relation to its effects on the target enzyme and on both M. bovis BCG and M. tuberculosis. The most potent triazole mimics the effects of deletion of the nat gene on growth, lipid disruption and intracellular survival. We also present the structure-activity relationship between NAT inhibition and effects on mycobacterial growth, and use ligand-protein analysis to give further insight into the structure-activity relationships. We conclude that screening a chemical library with NAT protein yields compounds that have high potential as anti-tubercular agents and that the inhibitors will allow further exploration of the biochemical pathway in which NAT is involved.
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford, OX1 3QT, UK.
authors.forename: Isaac M
authors.initials: IM
authors.lastname: Westwood
authors.forename: Sanjib
authors.initials: S
authors.lastname: Bhakta
authors.forename: Angela J
authors.initials: AJ
authors.lastname: Russell
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Fullam
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Anderton
authors.forename: Akane
authors.initials: A
authors.lastname: Kawamura
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Mulvaney
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Vickers
authors.forename: Veemal
authors.initials: V
authors.lastname: Bhowruth
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
authors.forename: Ajit
authors.initials: A
authors.lastname: Lalvani
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Davies
authors.forename: Edith
authors.initials: E
authors.lastname: Sim
mesh_terms: D000995:Antitubercular Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D057166:High-Throughput Screening Assays; D006801:Humans; D009163:Mycobacterium bovis / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D014230:Triazoles / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000995:Antitubercular Agents; D004791:Enzyme Inhibitors; D014230:Triazoles; D001191:Arylamine N-Acetyltransferase