An engineered prodrug selectively suppresses β-lactam resistant bacteria in a mixed microbial setting

Duda, A. M.; Ma, H. R.; Villalobos, C. A.; Kuhn, S. A.; He, K.; Seay, S. R.; Jackson, A. C.; Suh, C. M.; Puccio, E. A.; Anderson, D. A.; Fowler, V. G.; You, L.; Franz, K. J.

doi:10.1101/2024.08.02.606422

bioRxiv · 10.1101/2024.08.02.606422

An engineered prodrug selectively suppresses β-lactam resistant bacteria in a mixed microbial setting

Duda, A. M.·Ma, H. R.·Villalobos, C. A.·Kuhn, S. A.·He, K.·Seay, S. R.·Jackson, A. C.·Suh, C. M.·Puccio, E. A.·Anderson, D. A.·Fowler, V. G.·You, L.·Franz, K. J.

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Abstract

The rise of {beta}-lactam resistance necessitates new strategies to combat bacterial infections. We purposefully engineered the {beta}-lactam prodrug AcephPT to exploit {beta}-lactamase activity to selectively suppress resistant bacteria producing extended-spectrum-{beta}-lactamases (ESBLs). Selective targeting of resistant bacteria requires avoiding interaction with penicillin-binding proteins, the conventional targets of {beta}-lactam antibiotics, while maintaining recognition by ESBLs to activate AcephPT only in resistant cells. Computational approaches provide a rationale for structural modifications to the prodrug to achieve this biased activity. We show AcephPT selectively suppresses gram-negative ESBL-producing bacteria in clonal populations and in mixed microbial cultures, with effective selectivity for both lab strains and clinical isolates expressing ESBLs. Time-course NMR experiments confirm hydrolytic activation of AcephPT exclusively by ESBL-producing bacteria. In mixed microbial cultures, AcephPT suppresses proliferation of ESBL-producing strains while sustaining growth of {beta}-lactamase-non-producing bacteria, highlighting its potential to combat {beta}-lactam resistance while promoting antimicrobial stewardship. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=152 SRC="FIGDIR/small/606422v1_ufig1.gif" ALT="Figure 1"> View larger version (62K): org.highwire.dtl.DTLVardef@cd7f9aorg.highwire.dtl.DTLVardef@1747bc6org.highwire.dtl.DTLVardef@6a3419org.highwire.dtl.DTLVardef@825aa6_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Duda, A. M., Ma, H. R., Villalobos, C. A., Kuhn, S. A., He, K., Seay, S. R., Jackson, A. C., Suh, C. M., Puccio, E. A., Anderson, D. A., Fowler, V. G., You, L., Franz, K. J.. 2024-08-03. An engineered prodrug selectively suppresses β-lactam resistant bacteria in a mixed microbial setting. https://doi.org/10.1101/2024.08.02.606422

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