In vitro and in vivo combination of lytic phages and octapeptin OPX10053 against B-lactamase-producing clinical isolates of Klebsiella pneumoniae

Pacios, O.; Blasco, L.; Bleriot, I.; Fernandez-Garcia, L.; Lopez-Diaz, M.; Ortiz-Cartagena, C.; Barrio-Pujante, A.; Fernandez-Cuenca, F.; Aracil, B.; Oteo-Iglesias, J.; Hansford, K. A.; Tomas, M.

doi:10.1101/2023.03.15.532768

bioRxiv · 10.1101/2023.03.15.532768

In vitro and in vivo combination of lytic phages and octapeptin OPX10053 against B-lactamase-producing clinical isolates of Klebsiella pneumoniae

Pacios, O.·Blasco, L.·Bleriot, I.·Fernandez-Garcia, L.·Lopez-Diaz, M.·Ortiz-Cartagena, C.·Barrio-Pujante, A.·Fernandez-Cuenca, F.·Aracil, B.·Oteo-Iglesias, J.·Hansford, K. A.·Tomas, M.

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Abstract

Backgroundnovel approaches to treat Klebsiella pneumoniae infections are desperately needed, such as the use of rationally designed combination therapies. Objectivesto evaluate the in vitro and in vivo therapeutic potential of lytic phages against K. pneumoniae in combination with octapeptin, a promising class of lipopeptides with broad spectrum Gram-negative activity. Methodswe determined the MICs to twenty-two lipopeptide compounds and chose one octapeptin (OPX10053) for evaluation of potential synergism in combination with lytic phages using checkerboard assays, optical density growth curves and time-kill (CFU enumeration). Toxicity and efficacy in vivo assays were conducted on Galleria mellonella larvae. Resultsthis study reports the synergy found in vitro between the octapeptin OPX10053 and two lytic phages previously characterized by our research group (vB_KpnM-VAC13 and vB_KpnM-VAC66) against clinical isolates of K. pneumoniae. This synergy was validated by the FIC index, OD growth curves and time-kill assay when OPX10053 was added following 4 hours of phage exposure. Preliminary evaluation of toxicity revealed that OPX10053, even at subinhibitory concentrations and in phage combinations, exerts a toxic effect on larvae, which requires further investigation. ConclusionsThe in vitro application of lytic phages in combination with octapeptin OPX10053 showed synergistic activity. Exposure of G. mellonella to the lytic phages was well tolerated, whereas combination treatment with subinhibitory concentrations of OPX10053 did not attenuate toxicity. Even so, this innovative approach of combining lytic phages could open the door to some interesting associations between chemically synthesized drugs and biological entities. Sequential or simultaneous application alongside time, dosing and stewardship warrants further research.

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Pacios, O., Blasco, L., Bleriot, I., Fernandez-Garcia, L., Lopez-Diaz, M., Ortiz-Cartagena, C., Barrio-Pujante, A., Fernandez-Cuenca, F., Aracil, B., Oteo-Iglesias, J., Hansford, K. A., Tomas, M.. 2023-03-15. In vitro and in vivo combination of lytic phages and octapeptin OPX10053 against B-lactamase-producing clinical isolates of Klebsiella pneumoniae. https://doi.org/10.1101/2023.03.15.532768

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