Identification of novel scaffolds targeting Mycobacterium tuberculosis
Drug resistance in Mycobacterium tuberculosis is relentlessly progressing while only a handful of novel drug candidates are developed. Here we describe a GFP-based high-throughput screening of 386,496 diverse compounds to identify putative tuberculosis drug candidates. In an exploratory analysis of the model organism M. bovis BCG and M. smegmatis and the subsequent screening of the main library, we identified 6354 compounds with anti-mycobacterial activity. These hit compounds were predominantly selective for mycobacteria while dozens had activity in the low μM range. We tested toxicity against the human monocyte/macrophage cell line THP-1 and elaborated activity against M. tuberculosis growing in liquid broth, under unique conditions such as non-replicating persistence or inhibition of M. tuberculosis residing in macrophages. Finally, spontaneous compound-resistant M. tuberculosis mutants were selected and subsequently analyzed by whole genome sequencing. In addition to compounds targeting the well-described proteins Pks13 and MmpL3, we identified two novel scaffolds targeting the bifunctional guanosine pentaphosphate synthetase/ polyribonucleotide nucleotidyltransferase GpsI, or interacting with the aminopeptidase PepB, a probable pro-drug activator.
A newly identified scaffold targets the bifunctional enzyme GpsI.
The aminopeptidase PepB is interacting with a second novel scaffold.
Phenotypic screenings regularly identify novel compounds targeting Pks13 and MmpL3.
KeywordsMycobacterium tuberculosis High-throughput screening Early drug development GpsI PepB Pks13 MmpL3
We thank F. Hoffmann-La Roche AG (in particular Martin Brunner, Kenneth Bradley, and Paul Gillespie) for providing the compounds, helpful discussions, and comments on the manuscript. Additionally, we thank Karoline Wagner from the Institute of Medical Microbiology and Siricha Aluri from the Functional Genomics Center Zurich for sequencing the resistant mutants, and Chantal Quiblier and Erik C. Böttger for providing the S. aureus GFP-expressing plasmid and clinical isolates, respectively.
This work was supported by the University of Zurich and the Institue of Medical Microbiology. We achnolwedge financial support from Roche Extending the Innovation Network (EIN-UZ-14/0681), Swiss National Science Foundation (31003A_153349), Lungenliga Schweiz/Georg and Bertha Schwyzer-Winiker Stiftung (SLA-2018-02) and Baugarten Stiftung (STWF-18-011).
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