title: Antimicrobial and efflux pump inhibitory activity of caffeoylquinic acids from Artemisia absinthium against gram-positive pathogenic bacteria.
abstract: BACKGROUND
  Traditional antibiotics are increasingly suffering from the emergence of multidrug resistance amongst pathogenic bacteria leading to a range of novel approaches to control microbial infections being investigated as potential alternative treatments. One plausible antimicrobial alternative could be the combination of conventional antimicrobial agents/antibiotics with small molecules which block multidrug efflux systems known as efflux pump inhibitors. Bioassay-driven purification and structural determination of compounds from plant sources have yielded a number of pump inhibitors which acted against gram positive bacteria.
  
  
  RESULTS
  In this study we report the identification and characterization of 4',5'-O-dicaffeoylquinic acid (4',5'-ODCQA) from Artemisia absinthium as a pump inhibitor with a potential of targeting efflux systems in a wide panel of gram-positive human pathogenic bacteria. Separation and identification of phenolic compounds (chlorogenic acid, 3',5'-ODCQA, 4',5'-ODCQA) was based on hyphenated chromatographic techniques such as liquid chromatography with post column solid-phase extraction coupled with nuclear magnetic resonance spectroscopy and mass spectroscopy. Microbial susceptibility testing and potentiation of well know pump substrates revealed at least two active compounds; chlorogenic acid with weak antimicrobial activity and 4',5'-ODCQA with pump inhibitory activity whereas 3',5'-ODCQA was ineffective. These initial findings were further validated with checkerboard, berberine accumulation efflux assays using efflux-related phenotypes and clinical isolates as well as molecular modeling methodology.
  
  
  CONCLUSIONS
  These techniques facilitated the direct analysis of the active components from plant extracts, as well as dramatically reduced the time needed to analyze the compounds, without the need for prior isolation. The calculated energetics of the docking poses supported the biological information for the inhibitory capabilities of 4',5'-ODCQA and furthermore contributed evidence that CQAs show a preferential binding to Major Facilitator Super family efflux systems, a key multidrug resistance determinant in gram-positive bacteria.
authors.affiliation: Laboratory of Analytical Chemistry, University of Ioannina, Ioannina, Greece.
authors.forename: Yiannis C
authors.initials: YC
authors.lastname: Fiamegos
authors.forename: Panagiotis L
authors.initials: PL
authors.lastname: Kastritis
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Exarchou
authors.forename: Haley
authors.initials: H
authors.lastname: Han
authors.forename: Alexandre M J J
authors.initials: AM
authors.lastname: Bonvin
authors.forename: Jacques
authors.initials: J
authors.lastname: Vervoort
authors.forename: Kim
authors.initials: K
authors.lastname: Lewis
authors.forename: Michael R
authors.initials: MR
authors.lastname: Hamblin
authors.forename: George P
authors.initials: GP
authors.lastname: Tegos
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D031171:Artemisia absinthium / Q000737:chemistry*; D001599:Berberine / Q000378:metabolism; D018441:Biofilms / Q000187:drug effects; D018432:Drug Resistance, Multiple / Q000187:drug effects*; D004357:Drug Synergism; D006094:Gram-Positive Bacteria / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008958:Models, Molecular; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity; D011487:Protein Conformation; D011801:Quinic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity
chemical_list: D018528:ATP-Binding Cassette Transporters; D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D010936:Plant Extracts; C472707:caffeoylquinic acid; D011801:Quinic Acid; D001599:Berberine