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Chlorotonils exhibit potent activity against Mycobacterium tuberculosis, while resistance is mediated by MmpR5-MmpL5

Deschner, F.; Chengalroyen, M. D.; Ames, L.; Quach, D.; Aguilera Olvera, R.; Bosch, B.; Castro, A.; Kim, H.; Raman, K.; Thornton, N.; Wallach, J.; Rodrigues da Costa, F.; Allen, R.; Lupien, A.; Zuma, M.; Lynch, S.; Pogliano, J.; Sugie, J.; Rock, J. M.; Schnappinger, D.; Parish, T.; Mizrahi, V.; DeJesus, M.; Mueller, R.; Herrmann, J.

2025-12-10 microbiology
10.64898/2025.12.09.693342 bioRxiv
Show abstract

Treatment of Mycobacterium tuberculosis (Mtb) is challenging and requires administration of at least four different antibiotics. Unfortunately, multi-drug resistant Mtb strains continue to emerge, undermining the effectiveness of current treatment regimens and highlighting the urgent need for new therapeutics. In this study, we evaluated the potential of natural product-derived chlorotonils as anti-Mtb agents. We demonstrate that chlorotonils exhibit nanomolar potency against a range of attenuated and virulent Mtb strains. Mechanistic studies and resistance profiling in Mtb revealed that chlorotonils affect both lipid and energy metabolism. Through systems biology approaches, including the construction of an Mtb CRISPRi library specifically designed for chemical-genomic profiling, we identified MmpR5/MmpL5 as major driver of chlorotonil-resistance in Mtb leading also to cross-resistance with bedaquiline. Our findings highlight chlorotonils as valuable chemical tools to further dissect the role and function of the MmpS5-MmpL5 efflux pump in drug-resistant Mtb.

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