Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach

Antimicrobial resistance is a global health threat, which has been worsened by the slow development of new antibiotics. The rational design of natural-derived antimicrobial peptides (AMPs) offers a promising alternative for enhancing the efficacy of AMPs and accelerating drug discovery. This paper d...

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Main Authors: Silvana Aguilar, Daniel Moreira, Ana Laura Pereira Lourenço, Natalia Wilke, Matías A. Crosio, Andreanne Vasconcelos, Eder Alves Barbosa, Elizabete C. I. Bispo, Felipe Saldanha-Araujo, Marcelo H. S. Ramada, Franco M. Escobar, Cristina V. Torres, José R. S. A. Leite, Mariela M. Marani
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/15/3/449
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author Silvana Aguilar
Daniel Moreira
Ana Laura Pereira Lourenço
Natalia Wilke
Matías A. Crosio
Andreanne Vasconcelos
Eder Alves Barbosa
Elizabete C. I. Bispo
Felipe Saldanha-Araujo
Marcelo H. S. Ramada
Franco M. Escobar
Cristina V. Torres
José R. S. A. Leite
Mariela M. Marani
author_facet Silvana Aguilar
Daniel Moreira
Ana Laura Pereira Lourenço
Natalia Wilke
Matías A. Crosio
Andreanne Vasconcelos
Eder Alves Barbosa
Elizabete C. I. Bispo
Felipe Saldanha-Araujo
Marcelo H. S. Ramada
Franco M. Escobar
Cristina V. Torres
José R. S. A. Leite
Mariela M. Marani
author_sort Silvana Aguilar
collection DOAJ
description Antimicrobial resistance is a global health threat, which has been worsened by the slow development of new antibiotics. The rational design of natural-derived antimicrobial peptides (AMPs) offers a promising alternative for enhancing the efficacy of AMPs and accelerating drug discovery. This paper describes the rational design of improved peptide derivatives starting from hylin-Pul3, a peptide previously isolated from the frog <i>Boana pulchella,</i> by optimizing its hydrophobicity, cationicity, and amphipathicity. In silico screening identified six promising candidates: dHP3-31, dHP3-50, dHP3-50.137, dHP3-50.190, dHP3-84, and dHP3-84.39. These derivatives exhibited enhanced activity against Gram-negative bacteria, emphasizing the role of cationicity and the strategic arginine incorporation. Hemolytic assays revealed the derivatives’ improved selectivity, particularly for the derivatives with “imperfect amphipathicity”. In fibroblast assays, dHP3-84 was well-tolerated, while dHP3-84.39 promoted cell proliferation. Antioxidant assays (ABTS assays) highlighted the Trp-containing derivatives’ (dHP3-50.137, dHP3-31) significant activity. The lipid membrane interaction studies showed that hylin-Pul3 disrupts membranes directly, while dHP3-84.39, dHP3-50, and dHP3-50.137 promote vesicle aggregation. Conversely, dHP3-84 did not induce membrane disruption or aggregation, suggesting an intracellular mode of action. Machine learning models were effective in predicting bioactivity, as these predicted AMPs showed enhanced selectivity and potency. Among them, dHP3-84 demonstrated broad-spectrum potential. These findings highlight the value of rational design, in silico screening, and structure–activity studies in optimizing AMPs for therapeutic applications.
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spelling doaj-art-10db601667584226bbdd1995a0fdc8a32025-08-20T02:42:35ZengMDPI AGBiomolecules2218-273X2025-03-0115344910.3390/biom15030449Enhancing Antimicrobial Peptides from Frog Skin: A Rational ApproachSilvana Aguilar0Daniel Moreira1Ana Laura Pereira Lourenço2Natalia Wilke3Matías A. Crosio4Andreanne Vasconcelos5Eder Alves Barbosa6Elizabete C. I. Bispo7Felipe Saldanha-Araujo8Marcelo H. S. Ramada9Franco M. Escobar10Cristina V. Torres11José R. S. A. Leite12Mariela M. Marani13IPEEC-CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas, Puerto Madryn U9120ACD, ArgentinaResearch Center in Morphology and Applied Immunology, NuPMIA, Faculty of Medicine, University of Brasília, UnB, Brasília 70910-900, DF, BrazilPrograma de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Taguatinga 71966-700, DF, BrazilFacultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Universidad Nacional de Córdoba, Córdoba X5000HUA, ArgentinaFacultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Universidad Nacional de Córdoba, Córdoba X5000HUA, ArgentinaResearch Center in Morphology and Applied Immunology, NuPMIA, Faculty of Medicine, University of Brasília, UnB, Brasília 70910-900, DF, BrazilResearch Center in Morphology and Applied Immunology, NuPMIA, Faculty of Medicine, University of Brasília, UnB, Brasília 70910-900, DF, BrazilLaboratory of Hematology and Stem Cells, Faculty of Health Sciences, University of Brasília, UnB, Brasília 70910-900, DF, BrazilLaboratory of Hematology and Stem Cells, Faculty of Health Sciences, University of Brasília, UnB, Brasília 70910-900, DF, BrazilPrograma de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Taguatinga 71966-700, DF, BrazilDepartamento de Microbiología e Inmunología, Universidad Nacional de Río Cuarto, Ruta 36, Km 601, Río Cuarto 5800, ArgentinaDepartamento de Microbiología e Inmunología, Universidad Nacional de Río Cuarto, Ruta 36, Km 601, Río Cuarto 5800, ArgentinaResearch Center in Morphology and Applied Immunology, NuPMIA, Faculty of Medicine, University of Brasília, UnB, Brasília 70910-900, DF, BrazilIPEEC-CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas, Puerto Madryn U9120ACD, ArgentinaAntimicrobial resistance is a global health threat, which has been worsened by the slow development of new antibiotics. The rational design of natural-derived antimicrobial peptides (AMPs) offers a promising alternative for enhancing the efficacy of AMPs and accelerating drug discovery. This paper describes the rational design of improved peptide derivatives starting from hylin-Pul3, a peptide previously isolated from the frog <i>Boana pulchella,</i> by optimizing its hydrophobicity, cationicity, and amphipathicity. In silico screening identified six promising candidates: dHP3-31, dHP3-50, dHP3-50.137, dHP3-50.190, dHP3-84, and dHP3-84.39. These derivatives exhibited enhanced activity against Gram-negative bacteria, emphasizing the role of cationicity and the strategic arginine incorporation. Hemolytic assays revealed the derivatives’ improved selectivity, particularly for the derivatives with “imperfect amphipathicity”. In fibroblast assays, dHP3-84 was well-tolerated, while dHP3-84.39 promoted cell proliferation. Antioxidant assays (ABTS assays) highlighted the Trp-containing derivatives’ (dHP3-50.137, dHP3-31) significant activity. The lipid membrane interaction studies showed that hylin-Pul3 disrupts membranes directly, while dHP3-84.39, dHP3-50, and dHP3-50.137 promote vesicle aggregation. Conversely, dHP3-84 did not induce membrane disruption or aggregation, suggesting an intracellular mode of action. Machine learning models were effective in predicting bioactivity, as these predicted AMPs showed enhanced selectivity and potency. Among them, dHP3-84 demonstrated broad-spectrum potential. These findings highlight the value of rational design, in silico screening, and structure–activity studies in optimizing AMPs for therapeutic applications.https://www.mdpi.com/2218-273X/15/3/449hylinsin silico designamphipathicpeptide–membrane interactionscell selectivity
spellingShingle Silvana Aguilar
Daniel Moreira
Ana Laura Pereira Lourenço
Natalia Wilke
Matías A. Crosio
Andreanne Vasconcelos
Eder Alves Barbosa
Elizabete C. I. Bispo
Felipe Saldanha-Araujo
Marcelo H. S. Ramada
Franco M. Escobar
Cristina V. Torres
José R. S. A. Leite
Mariela M. Marani
Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
Biomolecules
hylins
in silico design
amphipathic
peptide–membrane interactions
cell selectivity
title Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
title_full Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
title_fullStr Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
title_full_unstemmed Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
title_short Enhancing Antimicrobial Peptides from Frog Skin: A Rational Approach
title_sort enhancing antimicrobial peptides from frog skin a rational approach
topic hylins
in silico design
amphipathic
peptide–membrane interactions
cell selectivity
url https://www.mdpi.com/2218-273X/15/3/449
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