Fosfomycin enhances the active transport of tobramycin in Pseudomonas aeruginosa.
Elevated levels of mucins present in bronchiectatic airways predispose patients to bacterial infections and reduce the effectiveness of antibiotic therapies by directly inactivating antibiotics. Consequently, new antibiotics that are not inhibited by mucins are needed to treat chronic respiratory infections caused by Pseudomonas aeruginosa and Staphylococcus aureus. In these studies, we demonstrate that fosfomycin synergistically enhances the activity of tobramycin in the presence of mucin. The bactericidal killing of a novel 4:1 (wt/wt) combination of fosfomycin-tobramycin (FTI) is superior (>9 log(10) CFU/ml) relative to its individual components fosfomycin and tobramycin. Additionally, FTI has a mutation frequency resulting in an antibiotic resistance >3 log(10) lower than for fosfomycin and 4 log(10) lower than for tobramycin for P. aeruginosa. Mechanistic studies revealed that chemical adducts are not formed, suggesting that the beneficial effects of the combination are not due to molecular modification of the components. FTI displayed time-kill kinetics similar to tobramycin and killed in a concentration-dependent fashion. The bactericidal effect resulted from inhibition of protein biosynthesis rather than cell wall biosynthesis. Studies using radiolabeled antibiotics demonstrated that tobramycin uptake was energy dependent and that fosfomycin enhanced the uptake of tobramycin in P. aeruginosa in a dose-dependent manner. Lastly, mutants resistant to fosfomycin and tobramycin were auxotrophic for specific carbohydrates and amino acids, suggesting that the resistance arises from mutations in specific active transport mechanisms. Overall, these data demonstrate that fosfomycin enhances the uptake of tobramycin, resulting in increased inhibition of protein synthesis and ultimately bacterial killing.
Gilead Sciences, Inc, Seattle, Washington, USA. Dave.MacLeod@gilead.com
David L
DL
MacLeod
Jyoti
J
Velayudhan
Thomas F
TF
Kenney
Joseph H
JH
Therrien
Jennifer L
JL
Sutherland
Lynn M
LM
Barker
William R
WR
Baker
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D004357:Drug Synergism; D005578:Fosfomycin / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D009077:Mucins / Q000378:metabolism / Q000494:pharmacology; D059645:Mutation Rate; D014176:Protein Biosynthesis; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D014031:Tobramycin / Q000378:metabolism / Q000494:pharmacology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D009077:Mucins; D005578:Fosfomycin; D014031:Tobramycin