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© Research
Publication : Scientific reports

Microspectrofluorimetry to dissect the permeation of ceftazidime in Gram-negative bacteria

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Scientific reports - 20 Apr 2017

Allam A, Maigre L, Vergalli J, Dumont E, Cinquin B, Alves de Sousa R, Pajovic J, Pinet E, Smith N, Herbeuval JP, Réfrégiers M, Artaud I, Pagès JM

Link to Pubmed [PMID] – 28428543

Sci Rep 2017 04;7(1):986

A main challenge in chemotherapy is to determine the in cellulo parameters modulating the drug concentration required for therapeutic action. It is absolutely urgent to understand membrane permeation and intracellular concentration of antibiotics in clinical isolates: passing the membrane barrier to reach the threshold concentration inside the bacterial periplasm or cytoplasm is the pivotal step of antibacterial activity. Ceftazidime (CAZ) is a key molecule of the combination therapy for treating resistant bacteria. We designed and synthesized different fluorescent CAZ derivatives (CAZ*, CAZ**) to dissect the early step of translocation-accumulation across bacterial membrane. Their activities were determined on E. coli strains and on selected clinical isolates overexpressing ß-lactamases. The accumulation of CAZ* and CAZ** were determined by microspectrofluorimetry and epifluorimetry. The derivatives were properly translocated to the periplasmic space when we permeabilize the outer membrane barrier. The periplasmic location of CAZ** was related to a significant antibacterial activity and with the outer membrane permeability. This study demonstrated the correlation between periplasmic accumulation and antibiotic activity. We also validated the method for approaching ß-lactam permeation relative to membrane permeability and paved the way for an original matrix for determining “Structure Intracellular Accumulation Activity Relationship” for the development of new therapeutic candidates.

https://www.ncbi.nlm.nih.gov/pubmed/28428543