CTX-1-Derived Cationic Peptides for Broad-Spectrum Low-Toxicity Killing

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Solution Overview

Problem

Existing antimicrobial peptides face challenges such as toxicity to eukaryotic cells, limited spectrum of activity, and instability in environmental conditions, hindering their therapeutic use against antibiotic-resistant bacteria, fungi, and viruses.

Innovation Solution

Development of cationic peptides derived from Naja atra cardiotoxin 1 (CTX-1) with a specific 20-amino acid structure (A-B-C-D-E-F-G) featuring alternating hydrophobic and charged regions, enhancing membrane penetration and stability, while minimizing toxicity and maintaining activity in high salt conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If existing antimicrobial peptides are used to treat antibiotic-resistant bacteria, then antimicrobial activity is achieved, but toxicity to eukaryotic cells increases

Engineering Contradiction:
Improveantimicrobial activityVSAvoidtoxicity to eukaryotic cells
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The peptide structure incorporates specific local features: a hydrophobic face for membrane insertion and a hydrophilic face with charged residues for electrostatic interaction with bacterial membranes. This local differentiation allows selective targeting of bacterial membranes while sparing eukaryotic cells, resolving the contradiction between antimicrobial activity and toxicity.

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The patent optimizes specific parameters including peptide length (15-30 residues), charge density (2-4 positive charges), and hydrophobicity ratio to achieve the optimal balance between antimicrobial efficacy and reduced eukaryotic toxicity. These parameter adjustments enable the peptide to effectively disrupt bacterial membranes while maintaining compatibility with eukaryotic cell membranes.

Inventive Principle:
Principle #35Parameter changes

2Adaptability or versatility

If antimicrobial peptides are designed with broad spectrum activity, then coverage against multiple pathogens is improved, but stability in environmental conditions deteriorates

Engineering Contradiction:
Improvespectrum of activityVSAvoidstability in environmental conditions
Core Design Contradiction:
Adaptability or versatilityVSStability of the object's composition

Solution Approach 1:

The peptide is divided into distinct functional segments: an N-terminal hydrophobic region for membrane anchoring, a central charged region for electrostatic interaction, and a C-terminal region for additional specificity. This segmentation allows each region to contribute to different aspects of activity and stability, enabling broad-spectrum coverage while maintaining environmental stability.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The peptide combines amino acids with different properties (hydrophobic, hydrophilic, charged, uncharged) in a specific arrangement to create a composite structure that simultaneously achieves broad-spectrum antimicrobial activity and environmental stability. The synergistic interaction of these diverse amino acid components resolves the contradiction between versatility and stability.

Inventive Principle:
Principle #40Composite materials

3Reliability

If peptide structure is optimized for membrane penetration, then antimicrobial efficacy is improved, but proteolytic stability decreases

Engineering Contradiction:
Improvemembrane penetration capabilityVSAvoidproteolytic stability
Core Design Contradiction:
ReliabilityVSStability of the object's composition

Solution Approach 1:

The design extracts only the essential membrane-penetrating features from complex natural antimicrobial peptides, creating a minimal sequence that retains penetration capability while removing vulnerable regions. This extraction approach maintains efficacy while reducing proteolytic susceptibility by eliminating unnecessary amino acid sequences.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

Instead of starting with a natural peptide and modifying it, the invention inverts the approach by designing a de novo peptide sequence that mimics the essential features of natural AMPs. This inversion allows optimization for both membrane penetration and proteolytic stability from the beginning, rather than trying to improve an existing structure.

Inventive Principle:
Principle #13The other way round (Inversion)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The peptides exhibit high antimicrobial, antiviral, and antifungal activity with low toxicity to eukaryotic cells, effective in various infection types, including those with high salt concentrations, and are proteolytically stable, making them suitable for diverse clinical applications.

Implementation Method 1

Bacterial membranes, unlike those eukaryotic, are rich in anionic phospholipids, such as phosphatidylserine and phosphatidylglycerol, which promotes the electrostatic interaction of the positively charged peptide with the membrane

Methodology Applied
Scientific EffectElectrostatic interaction: Electrostatics

Implementation Method 2

determining the structural perturbation of the lipid bilayer

Methodology Applied
Scientific EffectMembrane disruption:

Data Source

PatentEP3298025B1New amino acid sequences with microbicidal activity derived from naja atra cardiotoxin 1 (CTX-1)
Publication Date: 2026.02.11 UNIVERSITA DEGLI STUDI DI PARMA
  • EP3298025B1 patent drawingFigure 1A~2D
  • EP3298025B1 patent drawingFigure 3A~3B
  • EP3298025B1 patent drawingFigure 3C~4

AI summary

New linear peptides are described with broad spectrum of action, high antimicrobial antiviral and antifungal activities, combined with other characteristics of non-toxicity and persistence of activity in unfavorable environmental conditions. The peptides of the invention have a length of 20 amino acids and the structure: A-B-C-D-E-F-G, in which: A represents a basic amino acid; B, D, F represent respectively 5, 3 and 3 amino acids, chosen from the group of hydrophobic amino acids; C, E represent 3 amino acids, where each C and E comprises at least one basic amino acid and at least 1 amino acid forming hydrogen bonds; G represents 2 amino acids, chosen from basic amino acids and/or hydrophobic amino acids, their salts and/or mixtures thereof.