Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues
Methicillin-resistant <i>Staphylococcus pseudintermedius</i> (MRSP) constitutes an emerging health problem for companion animals in veterinary medicine. Therefore, discovery of novel antimicrobial agents for treatment of Staphylococcus-associated canine infections is urgently needed to r...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-03-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/24/6/1121 |
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| author | Ines Greco Johannes E. Hansen Bimal Jana Natalia Molchanova Alberto Oddo Peter W. Thulstrup Peter Damborg Luca Guardabassi Paul R. Hansen |
| author_facet | Ines Greco Johannes E. Hansen Bimal Jana Natalia Molchanova Alberto Oddo Peter W. Thulstrup Peter Damborg Luca Guardabassi Paul R. Hansen |
| author_sort | Ines Greco |
| collection | DOAJ |
| description | Methicillin-resistant <i>Staphylococcus pseudintermedius</i> (MRSP) constitutes an emerging health problem for companion animals in veterinary medicine. Therefore, discovery of novel antimicrobial agents for treatment of Staphylococcus-associated canine infections is urgently needed to reduce use of human antibiotics in veterinary medicine. In the present work, we characterized the antimicrobial activity of the peptoid <b>D2</b> against <i>S. pseudintermedius</i> and <i>Pseudomonas aeruginosa</i>, which is another common integumentary pathogen in dogs. Furthermore, we performed a structure–activity relationship study of <b>D2</b>, which included 19 peptide/peptoid analogs. Our best compound <b>D2D</b>, an all <span style="font-variant: small-caps;">d</span>-peptide analogue, showed potent minimum inhibitory concentrations (MICs) against canine <i>S. pseudintermedius</i> (2–4 µg/mL) and <i>P. aeruginosa</i> (4 µg/mL) isolates as well as other selected dog pathogens (2–16 µg/mL). Time–kill assays demonstrated that <b>D2D</b> was able to inhibit MRSP in 30 min at 1× MIC, significantly faster than <b>D2</b>. Our results suggest that at high concentrations <b>D2D</b> is rapidly lysing the bacterial membrane while <b>D2</b> is inhibiting macromolecular synthesis. We probed the mechanism of action at sub-MIC concentrations of <b>D2</b>, <b>D2D</b>, the <span style="font-variant: small-caps;">l</span>-peptide analog and its retro analog by a macromolecular biosynthesis assay and fluorescence spectroscopy. Our data suggest that at sub-MIC concentrations <b>D2D</b> is membrane inactive and primarily works by cell wall inhibition, while the other compounds mainly act on the bacterial membrane. |
| first_indexed | 2024-12-10T23:44:23Z |
| format | Article |
| id | doaj.art-ae1a1abc0ea446658bd0712e6600d5fe |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-10T23:44:23Z |
| publishDate | 2019-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-ae1a1abc0ea446658bd0712e6600d5fe2022-12-22T01:28:57ZengMDPI AGMolecules1420-30492019-03-01246112110.3390/molecules24061121molecules24061121Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide AnaloguesInes Greco0Johannes E. Hansen1Bimal Jana2Natalia Molchanova3Alberto Oddo4Peter W. Thulstrup5Peter Damborg6Luca Guardabassi7Paul R. Hansen8Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, DenmarkDepartment of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg C, DenmarkDepartment of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, DenmarkDepartment of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, DenmarkDepartment of Chemistry, University of Copenhagen, Universitetsparken 5, 2100 Copenhagen, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg C, DenmarkDepartment of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 4, 1870 Frederiksberg C, DenmarkDepartment of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen, DenmarkMethicillin-resistant <i>Staphylococcus pseudintermedius</i> (MRSP) constitutes an emerging health problem for companion animals in veterinary medicine. Therefore, discovery of novel antimicrobial agents for treatment of Staphylococcus-associated canine infections is urgently needed to reduce use of human antibiotics in veterinary medicine. In the present work, we characterized the antimicrobial activity of the peptoid <b>D2</b> against <i>S. pseudintermedius</i> and <i>Pseudomonas aeruginosa</i>, which is another common integumentary pathogen in dogs. Furthermore, we performed a structure–activity relationship study of <b>D2</b>, which included 19 peptide/peptoid analogs. Our best compound <b>D2D</b>, an all <span style="font-variant: small-caps;">d</span>-peptide analogue, showed potent minimum inhibitory concentrations (MICs) against canine <i>S. pseudintermedius</i> (2–4 µg/mL) and <i>P. aeruginosa</i> (4 µg/mL) isolates as well as other selected dog pathogens (2–16 µg/mL). Time–kill assays demonstrated that <b>D2D</b> was able to inhibit MRSP in 30 min at 1× MIC, significantly faster than <b>D2</b>. Our results suggest that at high concentrations <b>D2D</b> is rapidly lysing the bacterial membrane while <b>D2</b> is inhibiting macromolecular synthesis. We probed the mechanism of action at sub-MIC concentrations of <b>D2</b>, <b>D2D</b>, the <span style="font-variant: small-caps;">l</span>-peptide analog and its retro analog by a macromolecular biosynthesis assay and fluorescence spectroscopy. Our data suggest that at sub-MIC concentrations <b>D2D</b> is membrane inactive and primarily works by cell wall inhibition, while the other compounds mainly act on the bacterial membrane.https://www.mdpi.com/1420-3049/24/6/1121antimicrobialpeptidepeptoidmode of actioncanine infectionsmethicillin-resistant <i>Staphylococcus pseudintermedius</i><i>Pseudomonas aeruginosa</i> |
| spellingShingle | Ines Greco Johannes E. Hansen Bimal Jana Natalia Molchanova Alberto Oddo Peter W. Thulstrup Peter Damborg Luca Guardabassi Paul R. Hansen Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues Molecules antimicrobial peptide peptoid mode of action canine infections methicillin-resistant <i>Staphylococcus pseudintermedius</i> <i>Pseudomonas aeruginosa</i> |
| title | Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues |
| title_full | Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues |
| title_fullStr | Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues |
| title_full_unstemmed | Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues |
| title_short | Structure–Activity Study, Characterization, and Mechanism of Action of an Antimicrobial Peptoid D2 and Its <span style="font-variant: small-caps">d</span>- and <span style="font-variant: small-caps">l</span>-Peptide Analogues |
| title_sort | structure activity study characterization and mechanism of action of an antimicrobial peptoid d2 and its span style font variant small caps d span and span style font variant small caps l span peptide analogues |
| topic | antimicrobial peptide peptoid mode of action canine infections methicillin-resistant <i>Staphylococcus pseudintermedius</i> <i>Pseudomonas aeruginosa</i> |
| url | https://www.mdpi.com/1420-3049/24/6/1121 |
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