TY - JOUR
T1 - A novel tetrabranched antimicrobial peptide that neutralizes bacterial lipopolysaccharide and prevents septic shock in vivo
AU - Pini, Alessandro
AU - Falciani, Chiara
AU - Mantengoli, Elisabetta
AU - Bindi, Stefano
AU - Brunetti, Jlenia
AU - Iozzi, Sara
AU - Maria Rossolini, Gian
AU - Bracci, Luisa
PY - 2010/4
Y1 - 2010/4
N2 - We describe the nonnatural antimicrobial peptide KKIRVRLSA (M33) and its capacity to neutralize LPS-induced cytokine release, preventing septic shock in animals infected with bacterial species of clinical interest. M33 showed strong resistance to proteolytic degradation when synthesized in tetrabranched form with 4 peptides linked by a lysine core, making it suitable for use in vivo. HPLC and mass spectrometry demonstrated its stability in serum beyond 24 h. M33 was found to be very selective for gram-negative bacteria. Minimal inhibitory concentration (MIC) ranged from 0.3 to 3 μM for multidrug resistant clinical isolates of several pathogenic species, including Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii. M33 neutralized LPS derived from P. aeruginosa and K. pneumoniae, and prevented TNF-α release from LPS-activated macrophages, with an EC50 of 3.8e-8 M and 2.8e-7 M, respectively, as detected by sandwich ELISA. M33 activity was also tested in sepsis animal models. It averted septic shock symptoms due to Escherichia coli and P. aeruginosa in doses compatible with clinical use (5-25 mg/kg). These properties make tetrabranched M33 peptide a good candidate for the development of a new antibacterial drug.
AB - We describe the nonnatural antimicrobial peptide KKIRVRLSA (M33) and its capacity to neutralize LPS-induced cytokine release, preventing septic shock in animals infected with bacterial species of clinical interest. M33 showed strong resistance to proteolytic degradation when synthesized in tetrabranched form with 4 peptides linked by a lysine core, making it suitable for use in vivo. HPLC and mass spectrometry demonstrated its stability in serum beyond 24 h. M33 was found to be very selective for gram-negative bacteria. Minimal inhibitory concentration (MIC) ranged from 0.3 to 3 μM for multidrug resistant clinical isolates of several pathogenic species, including Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii. M33 neutralized LPS derived from P. aeruginosa and K. pneumoniae, and prevented TNF-α release from LPS-activated macrophages, with an EC50 of 3.8e-8 M and 2.8e-7 M, respectively, as detected by sandwich ELISA. M33 activity was also tested in sepsis animal models. It averted septic shock symptoms due to Escherichia coli and P. aeruginosa in doses compatible with clinical use (5-25 mg/kg). These properties make tetrabranched M33 peptide a good candidate for the development of a new antibacterial drug.
KW - Antibacterial
KW - Cystic fibrosis
KW - Dendrimers
KW - Gram-negative bacteria
KW - LPS
UR - http://www.scopus.com/inward/record.url?scp=77951626660&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77951626660&partnerID=8YFLogxK
U2 - 10.1096/fj.09-145474
DO - 10.1096/fj.09-145474
M3 - Article
C2 - 19917670
AN - SCOPUS:77951626660
VL - 24
SP - 1015
EP - 1022
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 4
ER -