Comparative analysis of internalisation, haemolytic, cytotoxic and antibacterial effect of membrane-active cationic peptides: aspects of experimental setup

• Published in: Amino acids Vol. 49; no. 6; pp. 1053 - 1067
• Main Authors: Horváti, Kata, Bacsa, Bernadett, Mlinkó, Tamás, Szabó, Nóra, Hudecz, Ferenc, Zsila, Ferenc, Bősze, Szilvia
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.06.2017; Springer Nature B.V
• Subjects: Analytical Chemistry; Anti-Bacterial Agents - chemical synthesis; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibacterial activity; Antibacterial materials; Antimicrobial agents; Antimicrobial Cationic Peptides - chemical synthesis; Antimicrobial Cationic Peptides - chemistry; Antimicrobial Cationic Peptides - pharmacology; Antimicrobial peptides; Biochemical Engineering; Biochemistry; Biomedical and Life Sciences; Cationic peptides; Cell Line, Tumor; Cell Membrane - chemistry; Cell Membrane - metabolism; Comparative analysis; Cytotoxicity; Drug delivery; Drug delivery systems; Drug development; Hemolysis - drug effects; Humans; Life Sciences; Minimum inhibitory concentration; Mycobacterium tuberculosis - growth & development; Neurobiology; Original Article; Peptides; Proteomics; Staphylococcus aureus - growth & development; Streptococcus infections; Tuberculosis; Haemolysis; Tuberculosis; Cell penetrating peptide; Antibacterial drug carrier; Cationic peptides; Antimicrobial peptide
• ISSN: 0939-4451; 1438-2199
• DOI: 10.1007/s00726-017-2402-9

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 HC 50 value. Therefore, these peptides and their analogues will be further investigated as drug delivery systems for antimicrobial agents.