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bioRxiv · 10.1101/2025.07.28.667204

Sphingosine synergizes with polymyxin antibiotics to kill Gram-negative bacteria

Mackinder, J. R.·Quinlan, M.·Wargo, M. J.
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Abstract

Antimicrobial resistance is an increasing threat to global health. However, there is a limited set of antibiotics that are effective against drug-resistant Gram-negative bacteria like Pseudomonas aeruginosa. One strategy to enhance the efficacy and longevity of existing antibiotics is by combining them with non-traditional antimicrobial adjuvants. Here, we examined if the host-derived antimicrobial lipid sphingosine could enhance the efficacy of a panel of antibiotics against P. aeruginosa in vitro. We found that sphingosine displayed strong synergy with the polymyxin antibiotics, polymyxin B and colistin, to inhibit growth of and kill P. aeruginosa, but did not significantly alter the efficacy of other tested antibiotic classes. The addition of sphingosine reduced the MIC of polymyxin B and colistin from 0.5 {micro}g/mL to 0.031 {micro}g/mL and 8 {micro}g/mL to 0.5 {micro}g/mL, respectively. This combination of sphingosine and polymyxin B synergized to inhibit the growth and survival of Klebsiella pneumoniae as well. In addition to sphingosine, we found that the sphingoid bases sphinganine (dihydrosphingosine) and phytosphingosine also enhanced the activity of polymyxins. Overall, these findings demonstrate that sphingosine is a potent adjuvant for polymyxins, and that the sphingosine- polymyxin combination is capable of killing P. aeruginosa and K. pneumoniae while using relatively low concentrations of polymyxin. This study may help in the development of new antimicrobial therapies for the treatment of Gram-negative bacterial infections. ImportanceAntibiotic resistance is an increasing threat to global health and there is a dire need to develop new therapies to treat multidrug-resistant infections. Here we show that sphingosine, a eukaryotic-derived antimicrobial lipid, synergizes with polymyxin antibiotics to inhibit the growth and survival of the Gram- negative bacteria, P. aeruginosa and K. pneumoniae. Thus, sphingosine has potential as an antimicrobial adjuvant for combinatorial therapies with polymyxins to treat Gram-negative infections.

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Mackinder, J. R., Quinlan, M., Wargo, M. J.. 2025-07-28. Sphingosine synergizes with polymyxin antibiotics to kill Gram-negative bacteria. https://doi.org/10.1101/2025.07.28.667204

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