Amphipathic Stapled Peptide for Intracellular Delivery
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Solution Overview
Problem
Conventional cell-penetrating peptides require micromolar concentrations to efficiently penetrate cells, limiting their effectiveness for intracellular delivery of substances, and existing stapled peptides composed mainly of hydrophobic amino acids may not be optimal for cell penetration.
Innovation Solution
Development of an amphipathic alpha-helical stapled peptide comprising hydrophobic and hydrophilic amino acids connected by covalent bonds, such as disulfide or carbon-carbon double bonds, to enhance cell-penetrating ability while maintaining chemical stability, using specific amino acid sequences and linkers to optimize penetration efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional cell-penetrating peptides are used, then cell penetration function is achieved, but high concentration (micromolar) is required reducing delivery efficiency
Solution Approach 1:
The invention changes the chemical structure parameters of the peptide by introducing staple motifs (covalent crosslinks between amino acid residues) to create a constrained alpha-helical structure. This structural parameter change enables the peptide to achieve high cell penetration at nanomolar concentrations rather than micromolar concentrations, directly resolving the contradiction between quantity required and delivery efficiency
Solution Approach 2:
The invention creates a composite peptide structure by combining hydrophobic amino acid residues (for membrane interaction) with hydrophilic amino acid residues (for solubility and stability), arranged in an amphipathic alpha-helical configuration with staple motifs. This composite structure achieves both high cell penetration ability and chemical stability at low concentrations
2Reliability
If stapled peptide with hydrophobic amino acids is used to regulate physiological activity, then protein-protein interaction is enhanced, but cell penetration ability is reduced
Solution Approach 1:
The invention applies local quality by creating distinct regions within the peptide: hydrophobic regions (with residues like Leu, Val, Phe, Trp) for protein-protein interactions and physiological activity regulation, and hydrophilic regions (with residues like Arg, Lys, His) for cell membrane interaction and penetration. The staple motifs are placed at specific local positions to maintain alpha-helical structure while enabling amphipathic character, thus resolving the contradiction between physiological activity and cell penetration
3Productivity
If dimeric peptide with 32 amino acids is used, then cell-penetrating ability is increased 500-fold, but peptide length increases reducing simplicity
Solution Approach 1:
The invention segments the long dimeric peptide (32 amino acids) into a shorter monomeric peptide (16 amino acids) by introducing staple motifs that create intramolecular crosslinks. These staples effectively divide the peptide chain into structured segments that maintain stability and function while reducing the overall length and complexity, achieving high cell penetration in a compact 16-residue structure
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 amphipathic alpha-helical stapled peptide achieves high cell-penetrating ability at lower concentrations compared to conventional peptides, effectively delivering biologically active substances into cells with improved stability and efficiency.
Implementation Method 1
enter cells by endocytotic mechanism
Implementation Method 2
easily recognize negatively charged substances on the cell surface
Implementation Method 3
two or more amino acids of the peptide are connected to each other by covalent linkages at two or more amino acid positions
Implementation Method 4
introducing cysteine instead of leucine into some hydrophobic residues of amphipathic peptides and connecting the peptides by two disulfide bonds
Implementation Method 5
amphipathic alpha-helical peptides comprising hydrophilic and hydrophobic amino acids
Implementation Method 6
amino acids playing an important role in protein-protein interactions consist mainly of hydrophobic functional groups
Data Source
AI summary
The present invention relates to a stapled peptide, a preparation method thereof and the use thereof, and more specifically to an amphipathic alpha-helical stapled peptide comprising hydrophobic amino acids and hydrophilic amino acids, a preparation method thereof, and the use thereof for intracellular delivery of an active substance.