With the rise in antibiotic resistance, new drugs are desperately needed against bacterial infections, especially tuberculosis (TB), a complex disease caused by <i>Mycobacterium tuberculosis</i> (Mtb) which kills 1.5 million people per year. Combining chemistry and genetics, we developed a new strategy called PROSPECT for rapidly identifying many new small molecule inhibitors of Mtb and for elucidating their mechanisms of action (MOA), by performing large-scale chemical screening on pooled genetic libraries containing 250-400 barcoded strains hypomorphic for individual essential genes. We created barcoded hypomorphic strains for 474 of the ~625 essential genes in Mtb and developed a multiplexed, whole-cell assay to measure strain abundance. Applying the approach with activity-enriched and unbiased libraries of &gt;100,000 compounds, we characterized &gt;25 million potential chemical-genetic interactions.<br/><br/>Using machine learning, we identified &gt;40 novel compounds against known MOAs, including new classes of inhibitors of DNA gyrase, mycolic acid biosynthesis, and folate biosynthesis. We also identified a pair of chemically distinct new inhibitors, BRD-9327 and BRD-8000, of a novel target, EfpA which are synergistic and display distinct interactions with EfpA. High-level resistance to one compound increases the sensitivity to and reduces the emergence of resistance to the others even when we applied antimicrobial monotherapy sequentially. Thus, the combination of BRD-9327 and BRD-8000 represents a proof-of-concept for the novel strategy of applying chemical-genetics to design of antimicrobial combination chemotherapy in which mutual collateral sensitivity is engineered and exploited.

Towards antimicrobial, anti-resistance chemotherapeutic strategies using large-scale chemical-genetics

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.