Chimeric Peptides Derived from Bovine Lactoferricin and Buforin II: Antifungal Activity against Reference Strains and Clinical Isolates of Candida spp.
Abstract
:1. Introduction
2. Results and Discussion
- (I)
- The C1 and C3 peptides were synthesized and evaluated in order to establish whether the antifungal activity is affected by the position of the RRWQWR or RLLR sequences in the chimera. Chimeras C2 and C4 contained Ahx (6-aminohexanoic acid residue) as a spacer for the two motifs. This is intended to establish whether the inclusion of the Ahx spacer between the two motifs affects the antifungal activity. The inclusion of the Ahx spacer facilitates the synthesis of the chimera and separates the two motifs so that each of them can interact independently with the cell surface of the pathogen.
- (II)
- Chimeras C5, C6, and C7 were synthesized and their antifungal activity was evaluated in order to determine whether partial or total replacement of Arg residues with Lys in RRWQWR and/or RLLR sequences affects their antifungal activity. Replacing Arg residues with Lys has been shown to facilitate and reduce the cost of chimeric synthesis [18].
- (III)
Antifungal Activity Against C. albicans Strains. µg/mL (µM) | ||||||
---|---|---|---|---|---|---|
Group | Code | Sequence | ATCC SC5314 | 256 HUSI-PUJ | ||
MIC | MFC | MIC | MFC | |||
Control | LfcinB (20–25) | RRWQWR | 200 (203) | 200 (203) | 200 (203) | 200 (203) |
BFII (32–35)Pal | RLLRRLLR | >200 (>183) | >200 (>183) | >200 (>183) | >200 (>183) | |
I | C1 | RRWQWRRLLR | 200 (131) | >200 (>131) | 100 (66) | >200 (>131) |
C2 | RRWQWR-Ahx-RLLR | 200 (122) | >200 (>122) | 200 (122) | >200(>122) | |
C3 | RLLRRRWQWR | 100 (66) | 200 (131) | 100 (66) | 200 (131) | |
C4 | RLLR-Ahx-RRWQWR | >200 (>122) | >200 (>122) | 200 (122) | >200 (>122) | |
II | C5 | RRWQWR-Ahx-KLLKKLLK | 100 (48) | 200 (97) | 100 (48) | 200 (97) |
C6 | KKWQWK-Ahx-RLLRRLLR | 50 (24) | 100 (48) | 50 (24) | 100 (48) | |
C7 | KKWQWK-Ahx-KLLKKLLK | 200 (101) | >200 (>101) | 200 (101) | >200 (>101) | |
III | C8 | (RRWQWR)2K-Ahx-RLLR | 100 (37) | 100 (37) | 100 (37) | 100 (37) |
C9 | (RRWQWR)2K-Ahx-RLLRRLLR | 50 (15) | 50 (15) | 50 (15) | 50 (15) |
2.1. Minimum Inhibitory and Fungicidal Concentration
2.2. Hemolytic Effect
2.3. Antifungal Activity of Chimeric Peptides Mixed with FLC
3. Materials and Methods
3.1. Reagents and Materials
3.2. Peptides
3.3. In Vitro Antifungal Susceptibility Test
3.4. Time-Kill Curves
3.5. Hemolysis Assays
3.6. Checkerboard Test
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Time-Kill Result against C. albicans Strains—µg/mL (µM) | ||||
---|---|---|---|---|
Peptide | ATCC SC5314 | 256 HUSI-PUJ | ||
Fungistatic * | Fungicide * | Fungistatic * | Fungicide * | |
C5 | 50 (24) | 100 (48) | 100 (48) | 200 (97) |
C6 | 25 (12) | 50 (24) | 50 (24) | 100 (48) |
C8 | 100 (37) | 200 (73) | 100 (37) | 200 (73) |
C9 | 13 (4) | 25 (8) | 13 (4) | 25 (8) |
Hemolytic Activity | ||
---|---|---|
Peptide | Concentration (μg/mL) * | % Hemolysis |
RLLR | 200 | 63 |
BFII (32–35)Pal | 200 | 2 |
LfcinB (20–25) | 200 | 1 |
C1 | 100–200 | 5 |
C2 | 200 | 3 |
C3 | 100–200 | 4 |
C4 | 200 | 5 |
C5 | 50–200 | 2 |
C6 | 25–100 | 2 |
C7 | 200 | 6 |
C8 | 100 | 2 |
C9 | 13–50 | 2–11 |
Synergistic Result C. albicans | ||||||||
---|---|---|---|---|---|---|---|---|
C. albicans Strain | Peptide | MICa | MICb | A | B | FICI | MICa/A | MICb/B |
ATCC SC5314 | C6 | 50 | 1 | 25 | 0.5 | 1 | 2 | 2 |
C8 | 200 | 0.5 | 25 | 0.13 | 0.38 | 8 | 4 | |
C9 | 100 | 0.5 | 25 | 0.13 | 0.5 | 4 | 4 | |
256 HUSI-PUJ | C6 | 100 | 32 | 3.1 | 32 | 1.03 | 32 | 1 |
C8 | 100 | 32 | 50 | 32 | 1.5 | 2 | 1 | |
C9 | 50 | 32 | 25 | 16 | 1 | 2 | 2 |
Synergistic Result C. glabrata and C. Auris | |||||||
---|---|---|---|---|---|---|---|
Strain | MICa | MICb | A | B | FICI | MICa/A | MICb/B |
C. glabrata ATCC 2001 | 100 | 0.3 | 50 | 0.03 | 0.6 | 2 | 10 |
C. glabrata 1875 CHU-PUJ | 400 | 4 | 12.5 | 4 | 1.03 | 32 | 1 |
C. auris 001 HUSI-PUJ | 400 | 32 | 12.5 | 16 | 0.53 | 32 | 2 |
C. auris 537 HUSI-PUJ | 400 | 64 | 200 | 32 | 1 | 2 | 2 |
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Aguirre-Guataqui, K.; Márquez-Torres, M.; Pineda-Castañeda, H.M.; Vargas-Casanova, Y.; Ceballos-Garzon, A.; Rivera-Monroy, Z.J.; García-Castañeda, J.E.; Parra-Giraldo, C.M. Chimeric Peptides Derived from Bovine Lactoferricin and Buforin II: Antifungal Activity against Reference Strains and Clinical Isolates of Candida spp. Antibiotics 2022, 11, 1561. https://doi.org/10.3390/antibiotics11111561
Aguirre-Guataqui K, Márquez-Torres M, Pineda-Castañeda HM, Vargas-Casanova Y, Ceballos-Garzon A, Rivera-Monroy ZJ, García-Castañeda JE, Parra-Giraldo CM. Chimeric Peptides Derived from Bovine Lactoferricin and Buforin II: Antifungal Activity against Reference Strains and Clinical Isolates of Candida spp. Antibiotics. 2022; 11(11):1561. https://doi.org/10.3390/antibiotics11111561
Chicago/Turabian StyleAguirre-Guataqui, Katherine, Mateo Márquez-Torres, Héctor Manuel Pineda-Castañeda, Yerly Vargas-Casanova, Andrés Ceballos-Garzon, Zuly Jenny Rivera-Monroy, Javier Eduardo García-Castañeda, and Claudia Marcela Parra-Giraldo. 2022. "Chimeric Peptides Derived from Bovine Lactoferricin and Buforin II: Antifungal Activity against Reference Strains and Clinical Isolates of Candida spp." Antibiotics 11, no. 11: 1561. https://doi.org/10.3390/antibiotics11111561