TY - JOUR
T1 - In vitro model of the Gram-negative bacterial cell envelope for investigation of anti-infective permeation kinetics
AU - Graef, Florian
AU - Richter, Robert
AU - Fetz, Verena
AU - Murgia, Xabier
AU - De Rossi, Chiara
AU - Schneider-Daum, Nicole
AU - Allegretta, Giuseppe
AU - Elgaher, Walid
AU - Haupenthal, Jörg
AU - Empting, Martin
AU - Beckmann, Felix
AU - Brönstrup, Mark
AU - Hartmann, Rolf
AU - Gordon, Sarah
AU - Lehr, Claus Michael
PY - 2018/8/10
Y1 - 2018/8/10
N2 - The cell envelope of Gram-negative bacteria is a formidable biological barrier, inhibiting the action of antibiotics by impeding their permeation into the intracellular environment. In-depth understanding of permeation through this barrier remains a challenge, despite its critical role in antibiotic activity. We therefore designed a divisible in vitro permeation model of the Gram-negative bacterial cell envelope, mimicking its three essential structural elements, the inner membrane and the periplasmic space as well as the outer membrane, on a Transwell setup. The model was characterized by contemporary imaging techniques and employed to generate reproducible quantitative and time-resolved permeation data for various fluorescent probes and anti-infective molecules of different structure and physicochemical properties. For a set of three fluorescent probes, the permeation through the overall membrane model was found to correlate with in bacterio permeation. Even more interestingly, for a set of six Pseudomonas quorum sensing inhibitors, such permeability data were found to be predictive for their corresponding in bacterio activities. Further exploration of the capabilities of the overall model yielded a correlation between the permeability of porin-independent antibiotics and published in bacterio accumulation data; a promising ability to provide structure-permeability information was also demonstrated. Such a model may therefore constitute a valuable tool for the development of novel anti-infective drugs.
AB - The cell envelope of Gram-negative bacteria is a formidable biological barrier, inhibiting the action of antibiotics by impeding their permeation into the intracellular environment. In-depth understanding of permeation through this barrier remains a challenge, despite its critical role in antibiotic activity. We therefore designed a divisible in vitro permeation model of the Gram-negative bacterial cell envelope, mimicking its three essential structural elements, the inner membrane and the periplasmic space as well as the outer membrane, on a Transwell setup. The model was characterized by contemporary imaging techniques and employed to generate reproducible quantitative and time-resolved permeation data for various fluorescent probes and anti-infective molecules of different structure and physicochemical properties. For a set of three fluorescent probes, the permeation through the overall membrane model was found to correlate with in bacterio permeation. Even more interestingly, for a set of six Pseudomonas quorum sensing inhibitors, such permeability data were found to be predictive for their corresponding in bacterio activities. Further exploration of the capabilities of the overall model yielded a correlation between the permeability of porin-independent antibiotics and published in bacterio accumulation data; a promising ability to provide structure-permeability information was also demonstrated. Such a model may therefore constitute a valuable tool for the development of novel anti-infective drugs.
KW - anti-infectives
KW - drug delivery
KW - drug design
KW - Gram-negative bacterial cell envelope
KW - in vitro model
UR - http://www.scopus.com/inward/record.url?scp=85047061773&partnerID=8YFLogxK
U2 - 10.1021/acsinfecdis.7b00165
DO - 10.1021/acsinfecdis.7b00165
M3 - Article
C2 - 29750862
AN - SCOPUS:85047061773
SN - 2373-8227
VL - 4
SP - 1188
EP - 1196
JO - ACS Infectious Diseases
JF - ACS Infectious Diseases
IS - 8
ER -