Comparative analysis of internalisation, haemolytic, cytotoxic and antibacterial effect of membrane-active cationic peptides: aspects of experimental setup.
Cationic peptides proved fundamental importance as pharmaceutical agents and/or drug carrier moieties functioning in cellular processes. The comparison of the in vitro activity of these peptides is an experimental challenge and a combination of different methods, such as cytotoxicity, internalisation rate, haemolytic and antibacterial effect, is necessary. At the same time, several issues need to be addressed as the assay conditions have a great influence on the measured biological effects and the experimental setup needs to be optimised. Therefore, critical comparison of results from different assays using representative examples of cell penetrating and antimicrobial peptides was performed and optimal test conditions were suggested. Our main goal was to identify carrier peptides for drug delivery systems of antimicrobial drug candidates. Based on the results of internalisation, haemolytic, cytotoxic and antibacterial activity assays, a classification of cationic peptides is advocated. We found eight promising carrier peptides with good penetration ability of which Penetratin, Tat, Buforin and Dhvar4 peptides showed low adverse haemolytic effect. Penetratin, Transportan, Dhvar4 and the hybrid CM15 peptide had the most potent antibacterial activity on Streptococcus pneumoniae (MIC lower than 1.2 μM) and Transportan was effective against Mycobacterium tuberculosis as well. The most selective peptide was the Penetratin, where the effective antimicrobial concentration on pneumococcus was more than 250 times lower than the HC50value. Therefore, these peptides and their analogues will be further investigated as drug delivery systems for antimicrobial agents.
MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Pázmány P. stny. 1/A, Budapest, 1117, Hungary.
Kata
0000-0003-4092-6011
K
Horváti
MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Pázmány P. stny. 1/A, Budapest, 1117, Hungary.
Bernadett
B
Bacsa
Laboratory of Bacteriology, Korányi National Institute for Tuberculosis and Respiratory Medicine, Budapest, Hungary.
Tamás
T
Mlinkó
Laboratory of Bacteriology, Korányi National Institute for Tuberculosis and Respiratory Medicine, Budapest, Hungary.
Nóra
N
Szabó
MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Pázmány P. stny. 1/A, Budapest, 1117, Hungary.|Department of Organic Chemistry, Eötvös Loránd University, Budapest, Hungary.
Ferenc
F
Hudecz
Biomolecular Self-Assembly Group, Research Centre for Natural Sciences, Institute of Materials and Environmental Chemistry, Hungarian Academy of Sciences, Budapest, Hungary.
Ferenc
F
Zsila
MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Pázmány P. stny. 1/A, Budapest, 1117, Hungary. szilvia.bosze@gmail.com.
Szilvia
S
Bősze
D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000254:growth & development*; D013211:Staphylococcus aureus / Q000254:growth & development*
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides
Antibacterial drug carrier; Antimicrobial peptide; Cationic peptides; Cell penetrating peptide; Haemolysis; Tuberculosis