Antibiotic-based conjugates containing antimicrobial HLopt2 peptide: design, synthesis, antimicrobial and cytotoxic activities

Natalia Ptaszyńska , Katarzyna Gucwa , Katarzyna Olkiewicz , Anna Łęgowska , Joanna Okońska , Jarosław Ruczyński , Agata Gitlin-Domagalska , Dawid Dębowski , Sławomir Milewski , Krzysztof Rolka

Abstract

Recent studies have shown that modified human lactoferrin 20-31 fragment, named HLopt2, possesses antibacterial and antifungal activity. Thus, we decided to synthesize and evaluate the biological activity of a series of conjugates based on this peptide and one of the antimicrobials with proven antibacterial (ciprofloxacin, CIP, and levofloxacin, LVX) or antifungal (fluconazole, FLC) activity. The drugs were covalently connected to the peptide via amide, methylenecarbonyl moieties, or a disulfide bridge. The antibacterial and antifungal activities were evaluated under Clinical and Laboratory Standard Institute (CLSI) recommended conditions or in a low-salt brain-heart infusion diluted medium (BHI1/100). Results showed that conjugation of the peptide with the drug increased its antimicrobial activity up to 4-fold. Under CLSI-recommended conditions, all the compounds revealed rather low efficiency. Among conjugates, the highest antibacterial activity was recorded for the CIP-Cys-S-S-HLopt2-NH2 (III). In BHI1/100, which had lower differentiating properties, all of the conjugates revealed low MIC and MMC (minimum inhibitory and microbicidal concentrations) values. The disulfide bridge used as a linker in the most active conjugate (III) upon incubation with S. aureus cells is reduced, releasing constituent peptide and CIP-Cys. In addition, we showed that its fluorescently labeled analogue and constituent peptide are able to be internalized into both C. albicans and S. aureus cells. Moreover, the invaluable advantage of the presented conjugates was their low toxicity to mammalian cells and very low hemolytic activity. The current research can form a solid basis for further in vivo studies and drug development.
Author Natalia Ptaszyńska (FCh / DMB / LBCh)
Natalia Ptaszyńska,,
- Laboratory of Bioorganic Chemistry
, Katarzyna Gucwa (FCh / DMB) - [Politechnika Gdańska]
Katarzyna Gucwa,,
- Department of Molecular Biochemistry
- Politechnika Gdańska
, Katarzyna Olkiewicz (FCh / DMB / LBCh)
Katarzyna Olkiewicz ,,
- Laboratory of Bioorganic Chemistry
, Anna Łęgowska (FCh / DMB / LBCh)
Anna Łęgowska,,
- Laboratory of Bioorganic Chemistry
, Joanna Okońska (FCh / DMB / LBCh)
Joanna Okońska,,
- Laboratory of Bioorganic Chemistry
, Jarosław Ruczyński (FCh / DMB / LChBAC)
Jarosław Ruczyński,,
- Laboratory of Chemistry of Biologically Active Compounds
, Agata Gitlin-Domagalska (FCh / DMB / LBCh)
Agata Gitlin-Domagalska,,
- Laboratory of Bioorganic Chemistry
, Dawid Dębowski (FCh / DMB / LBCh)
Dawid Dębowski,,
- Laboratory of Bioorganic Chemistry
, Sławomir Milewski - [Politechnika Gdanska]
Sławomir Milewski,,
-
- Politechnika Gdanska
, Krzysztof Rolka (FCh / DMB / LBCh)
Krzysztof Rolka,,
- Laboratory of Bioorganic Chemistry
Journal seriesACS Chemical Biology, ISSN 1554-8929, e-ISSN 1554-8937, (N/A 100 pkt)
Issue year2019
Vol14
No10
Pages2233-2242
Publication size in sheets0.5
Keywords in Polishkoniugaty peptydowe, antybiotykipeptydy przeciwdrobnoustrojowe, reakcja click, miedzycząsteczkowe mostki disulfidowe, analog laktoferrycyny, aktywność przeciwdrobnoustrojowa
Keywords in Englishpeptide conjugates, antibiotics, antimicrobial peptide, click chemistry, intermolecular disulfide bond, lactoferricin analogue, fungicidal, antimicrobial activity
ASJC Classification2700 General Medicine; 1303 Biochemistry; 1313 Molecular Medicine
Abstract in PolishNa drodze syntezy chemicznej otrzymano szereg koniugatów, każdy skadał się z farmaceutyku o aktywności przeciwbakteryjnej(cioprofloksacycyny,lewofloksacyny)lub przeciwgrzybowej(flukonazol)oraz peptydu (HLopt2) o aktywności przeciwdrobnoustrojowej. Oba komponenty połączone były wiązaniami kowalencyjnymi. Najwyższą aktywnością przeciwbakteryjną wykazał koniugat, w którym oba komponenty połączone były mostkiem disulfidowym. Znaczony fluorescencyjnie analog tego koniugatu ulegał internalizacji do komórek bakteryjnych i grzybowych. Syntetyzowane związki wykazały niską aktywność hemolityczną i cytotoksyczną w stosunku do zdrowych komórek ludzkich.
DOIDOI:10.1021/acschembio.9b00538
URL https://pubs.acs.org/doi/pdf/10.1021/acschembio.9b00538
Languageen angielski
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Ptaszynska_Natalia_Antibiotic-Based_Conjugates_2019.pdf 3.45 MB
Score (nominal)100
Score sourcejournalList
ScoreMinisterial score = 100.0, 03-04-2020, ArticleFromJournal
Publication indicators WoS Citations = 0; Scopus Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2018 = 1.074; WoS Impact Factor: 2018 = 4.374 (2) - 2018=4.698 (5)
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