Novel antimicrobial peptides based on Protegrin-1: In silico and in vitro assessments

Microb Pathog. 2024 Nov:196:106931. doi: 10.1016/j.micpath.2024.106931. Epub 2024 Sep 15.

Abstract

The development of antibiotic resistance has caused significant health problems. Antimicrobial peptides (AMPs) are considered next-generation antibiotics. Protegrin-1 (PG-1) is a β-hairpin AMP with a membrane-binding capacity. This study used twelve PG-1 analogs with different amino acid substitutions. Coarse-grained molecular dynamics (MD) simulations were used to assess these analogs, and their physicochemical properties were computed using the Antimicrobial Peptide Database. Three AMPs, PEP-D, PEP-C, and PEP-H, were chosen and synthesized for antibacterial testing. The microbroth dilution technique and hemolytic assays evaluated the antimicrobial efficacy and cellular toxicity. The checkerboard method was used to test the combined activity of AMP and standard antibiotics. Cell membrane permeability and electron microscopy were used to evaluate the mode of action. The chemical stability of the selective AMP, PEP-D, was assessed by a validated HPLC method. PEP-D consists of 16-18 amino acid residues and has a charge of +7 and a hydrophobicity of 44 %, similar to PG-1. It can efficiently inactivate bacteria by disrupting cell membranes and significantly reducing hemolytic activity. Chemical stability studies indicated that AMP was stable at 40 °C for six months under autoclave conditions. This study could introduce the potential therapeutic application of selective AMP as an anti-infective agent.

Keywords: Antibacterial resistance; Antimicrobial peptides; Membrane disruption; Molecular dynamics; Physicochemical properties; Protegrin-1; Stability.

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / pharmacology
  • Antimicrobial Cationic Peptides* / chemistry
  • Antimicrobial Cationic Peptides* / pharmacology
  • Antimicrobial Peptides / chemistry
  • Antimicrobial Peptides / pharmacology
  • Bacteria / drug effects
  • Cell Membrane / drug effects
  • Cell Membrane Permeability / drug effects
  • Computer Simulation
  • Erythrocytes / drug effects
  • Hemolysis* / drug effects
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Microbial Sensitivity Tests*
  • Molecular Dynamics Simulation*

Substances

  • protegrin-1
  • Antimicrobial Cationic Peptides
  • Anti-Bacterial Agents
  • Antimicrobial Peptides