pH-Responsive Lipid Assemblies for Gene Delivery

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Solution Overview

Problem

Current non-viral gene delivery systems, such as cationic liposomes, face challenges with colloidal stability and toxicity, and while amphoteric liposomes offer improved tolerance, they lag behind viral systems in transfection efficacy and immune response issues.

Innovation Solution

Development of lipids with transfection enhancer elements (TEEs) that undergo a pH-sensitive hydrophile-hydrophobe transition, enhancing fusogenicity with cell membranes, and their incorporation into lipid assemblies like liposomes to improve cellular uptake and delivery of nucleic acids.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If cationic liposomes are used for gene delivery, then transfection efficiency is improved, but colloidal stability deteriorates and toxicity increases

Engineering Contradiction:
Improvetransfection efficiencyVSAvoidcolloidal stability
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying the charge state of liposomes as a function of pH. The liposomes are designed to be cationic at acidic pH (enhancing transfection efficiency) and anionic at physiological pH (improving colloidal stability and reducing toxicity). This dynamic parameter change allows the system to optimize performance at different stages of the delivery process.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention implements dynamics through pH-responsive charge switching. The liposome charge is not fixed but dynamically changes in response to pH variations encountered during circulation and cellular uptake. This dynamic adaptation enables the system to maintain stability in blood (anionic at pH 7.4) while becoming fusogenic at endosomal pH (cationic at acidic conditions).

Inventive Principle:
Principle #15Dynamics

2Productivity

If cationic liposomes are used for gene delivery, then transfection efficiency is improved, but toxicity increases

Engineering Contradiction:
Improvetransfection efficiencyVSAvoidtoxicity
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent applies parameter changes by modifying the charge state of liposomes as a function of pH. The liposomes are designed to be cationic at acidic pH (enhancing transfection efficiency) and anionic at physiological pH (improving colloidal stability and reducing toxicity). This dynamic parameter change allows the system to optimize performance at different stages of the delivery process.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention implements dynamics through pH-responsive charge switching. The liposome charge is not fixed but dynamically changes in response to pH variations encountered during circulation and cellular uptake. This dynamic adaptation enables the system to maintain stability in blood (anionic at pH 7.4) while becoming fusogenic at endosomal pH (cationic at acidic conditions).

Inventive Principle:
Principle #15Dynamics

3Object-affected harmful factors

If amphoteric liposomes are used for gene delivery, then safety and tolerance are improved, but transfection efficacy deteriorates

Engineering Contradiction:
Improvesafety and toleranceVSAvoidtransfection efficacy
Core Design Contradiction:
Object-affected harmful factorsVSProductivity

Solution Approach 1:

The patent applies dynamics through pH-responsive charge switching. The liposome charge is not fixed but dynamically changes in response to pH variations encountered during circulation and cellular uptake. This dynamic adaptation enables the system to maintain stability in blood (anionic at pH 7.4) while becoming fusogenic at endosomal pH (cationic at acidic conditions).

Inventive Principle:
Principle #15Dynamics

Solution Approach 2:

The invention utilizes phase transitions in the form of charge state transitions driven by pH changes. The liposomes undergo a charge phase transition from anionic to cationic as they move from physiological pH to acidic endosomal pH, enabling them to switch between stable circulation and active membrane fusion states.

Inventive Principle:
Principle #36Phase transitions

4Productivity

If viral vectors are used for gene delivery, then transfection efficacy is improved, but immune response increases

Engineering Contradiction:
Improvetransfection efficacyVSAvoidimmune response
Core Design Contradiction:
ProductivityVSObject-generated harmful factors

Solution Approach 1:

The patent applies the extraction principle by removing the viral components that trigger immune responses while retaining and enhancing the essential fusogenic function. The invention uses synthetic pH-responsive lipids that mimic the pH-dependent membrane fusion capability of viral envelopes without containing viral genetic material or surface proteins, thereby eliminating immunogenicity while maintaining transfection efficacy.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention employs synthetic, non-viral lipid structures that can be designed for single-use delivery without the long-term persistence and immune recognition associated with viral vectors. These synthetic liposomes provide the necessary fusogenic function temporarily during delivery then are metabolized without triggering sustained immune responses.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

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 pH-sensitive lipids enhance the fusogenicity and cellular uptake of liposomes, improving the delivery of nucleic acids and reducing toxicity, offering a safer and more effective non-viral gene delivery method compared to existing systems.

Implementation Method 1

The TEE comprises a hydrophobic moiety and a pH sensitive hydrophilic moiety which responds to a pH drop from physiological pH to low pH by a hydrophile-hydrophobe transition

Methodology Applied
Scientific EffectpH-sensitive hydrophile-hydrophobe transition: Phase Change

Implementation Method 2

the pH sensitive hydrophilic moiety of each TEE is independently a weak acid having pKa of between 2 and 6

Methodology Applied
Scientific EffectProtonation:

Data Source

PatentEP2125031B1Lipids and lipid assemblies comprising transfection enhancer elements
Publication Date: 2017.11.01 MARINA BIOTECH INC
  • EP2125031B1 patent drawingFigure 1
  • EP2125031B1 patent drawingFigure 2
  • EP2125031B1 patent drawingFigure 3

AI summary

Lipid assemblies, such as liposomes, comprising transfection enhancer elements (TEE's), which are complexed with the lipid assemblies by means of ionic interactions, or lipids incorporating such TEE's are disclosed for enhancing the fusogenicity of the lipid assemblies. The TEE's have the formula: hydrophobic moiety - pH sensitive hydrophilic moiety (II) The pH sensitive hydrophilic moiety of each TEE is a weak acid having a pka of between 2 and 6 or a zwitterionic structure comprising a combination of acidic groups with weak bases having a pKa of between 3 and 8. Lipids incorporating one or more such TEEs have the formula (I): Lipid moiety- [Hydrophobic moiety - pH sensitive hydrophilic moiety] (I).