Cell-Penetrating Peptide Constructs for pH-Triggered Endosomal Release
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing cell penetrating peptides (CPPs) face limitations in efficiently delivering cargo, such as siRNA, across lipid membranes due to heterogeneity, low stability, and premature dissociation, often resulting in endosomal trapping and limited intracellular delivery.
Innovation Solution
A membrane-permeable construct comprising a cell penetrating amino acid sequence with modifications, including histidine residues and a fatty acid chain, exhibits pH sensitivity for efficient endosomal release and targeted delivery, particularly effective in low pH cancer microenvironments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If non-covalent cell penetrating amino acid sequence/cargo complexation is used, then simple complex formation with high mixing versatility is achieved, but heterogeneity and low stability of complexes occur, and premature or limited dissociation of cargo from complexes happens
Solution Approach 1:
The patent modifies the cell penetrating peptide parameters by incorporating specific amino acid sequences with defined properties (e.g., arginine-rich sequences, specific hydrophobic residues) to optimize the balance between complex formation ease and stability. The peptide length, charge density, and hydrophobicity are tuned to achieve stable complexes that resist premature dissociation while maintaining simple formation protocols.
2Strength
If cell penetrating amino acid sequences with both hydrophilic and hydrophobic regions are designed, then cell membrane interaction is enhanced, but endosomal trapping and limited intracellular delivery occur
Solution Approach 1:
The patent exploits pH-induced phase transitions of the cell penetrating peptide. At endosomal pH (acidic conditions), the peptide undergoes conformational changes or protonation that triggers endosomal escape, releasing the cargo into the cytoplasm. This phase transition mechanism allows strong membrane interaction at neutral pH while enabling efficient intracellular delivery through pH-responsive release in acidic endosomes.
Solution Approach 2:
The peptide structure is designed to be dynamic rather than static, allowing conformational adjustments in response to environmental pH changes. This dynamic behavior enables the peptide to adapt its membrane interaction strength and cargo release properties based on the local pH environment, overcoming endosomal trapping.
3Productivity
If existing cell penetrating peptides are used for cargo delivery, then cargo transport across lipid membrane is achieved, but heterogeneity, low stability, and premature dissociation limit effective delivery
Solution Approach 1:
The patent creates composite structures by combining cell penetrating peptide sequences with specific cargo molecules (e.g., siRNA, oligonucleotides) to form stable conjugates or complexes. The composite design integrates the cargo-binding properties of the peptide with the functional properties of the cargo, achieving both stable complexation and efficient intracellular delivery while reducing heterogeneity through defined stoichiometry and 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 construct achieves enhanced siRNA delivery with improved stability and reduced toxicity, offering potential for targeted cancer therapy and effective gene silencing.
Implementation Method 1
They form a non-covalent complex mainly via electrostatic interactions
Implementation Method 2
although hydrophobic interactions may play a role in the stability of the formed complexes
Implementation Method 3
the constructs of the present invention have good pH sensitivity and are good at delivery of cargo into the intracellular compartment. In particular, the constructs are sensitive to changes in pH which allows for efficient cell delivery of cargo via endosomes
Data Source
AI summary
The present invention provides for a membrane-permeable construct for transport of cargo across a lipid membrane and subsequent delivery of cargo into cells, wherein the construct comprises a cell penetrating amino acid sequence and a fatty acid chain attached to the N terminus thereof, wherein the cell penetrating amino acid sequence comprises the sequence of SEQ ID NO: 1, or comprises a modified sequence of SEQ ID NO: 1, wherein the cell penetrating amino acid sequence comprises two or more histidine residues by substitution to its N-terminal part and/or addition to its N-terminus, and wherein the cell penetrating amino acid sequence is optionally chemically modified at the C terminus.