Synthese en toepassing van een lipofiel aminozuurderivaat voor het verbeteren van de membraanaffiniteit van antimicrobiële peptiden
Slootweg, Jack C.; Van Schaik, Timo B.; Van Ufford, H. C. Quarles; Breukink, Eefjan; Liskamp, Rob M. J.; Rijkers, DirkT. S.
(2014) Pharmaceutisch Weekblad, volume 149, issue 43, pp. 174 - 179
(Article)
Abstract
OBJECTIVE: To increase the potential of membrane-acting peptides as possible novel drug-like compounds by increasing lipophilicity and thereby enhancing membrane affinity. DESIGN: An Fmoc-protected enantiomerically pure lipophilic amino acid (Fmoc-Lad-OH), which contains a nine carbon atom hydrophobic side chain, was designed. Fmoc-Lad-OH can be introduced into any peptide sequence using
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standard solid phase peptide synthesis to increase the lipophilicity of a peptide without sacrificing important polar segments of a peptide like for instance the N and C-termini. The antimicrobial decapeptide anoplin was chosen as a model peptide to test the hypothesis. METHODS: Fmoc-Lad-OH was prepared via organic synthesis and incorporated into the anoplin peptide sequence using solid-phase peptide synthesis followed by reversed-phase HPLC purification. Biological activity was evaluated using microtiter dilution bacterial growth assays, haemolytic assays and membrane vesicle leakage experiments. RESULTS: All three lipophilic analogues show a dramatic increase in antimicrobial activity: up to 4-8 times better for Escherichia coli (Gram-negative] and over one order of magnitude for Staphylococcus aureus (Gram-positive) compared to anoplin. Although the haemolytic activity was increased for the lipophilic analogues, the concentration at which 50% lysis will occur (EC50) was still one order of magnitude higher than the determined MICs. In the membrane vesicle leakage experiments the lipophilic analogues showed a higher lytic activity than anoplin, in agreement with the observed MIC values. CONCLUSION: Introduction of Lad into anoplin clearly showed a positive effect, which suggests that Fmoc-Lad-OH could be used as a general approach to increase membrane affinity of membrane-acting peptides.
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Keywords: amino acid derivative, polypeptide antibiotic agent, amino acid sequence, amino acid synthesis, amino terminal sequence, antimicrobial activity, article, assay, carboxy terminal sequence, dilution, EC50, Escherichia coli, hemolysis, hemolysis assay, hydrophobicity, lipophilicity, membrane vesicle, microtiter dilution, minimum inhibitory concentration, nonhuman, peptide synthesis, pH, reversed phase high performance liquid chromatography, Staphylococcus aureus
ISSN: 0031-6911
Publisher: De Koninklijke Nederlandse Maatschappij ter bevordering der Pharmacie
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