Peptide Shuttle Agents for Cytosolic Delivery
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current cell delivery technologies face challenges in efficiently transporting large polypeptide cargos into eukaryotic cells, particularly due to endosomal trapping and safety concerns associated with existing methods, which limits their therapeutic potential in fields like cell therapy.
Innovation Solution
Development of synthetic peptides comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), optionally with a histidine-rich domain, to enhance the transduction efficiency of polypeptide cargos into the cytosol of target eukaryotic cells without being covalently bound to the cargo.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If existing cell delivery technologies are used to transport large polypeptide cargos into eukaryotic cells, then some delivery is achieved, but transduction efficiency is limited due to endosomal trapping
Solution Approach 1:
The invention divides the shuttle agent into two separate functional domains: an endosome leakage domain (ELD) and a cell penetrating domain (CPD). These domains can be used independently or in combination, allowing optimized delivery strategies. The ELD specifically addresses endosomal escape while the CPD handles cellular uptake, resolving the contradiction by segmenting the delivery function into specialized components.
Solution Approach 2:
The invention creates composite shuttle agents by operably linking the ELD and CPD domains to form multi-domain peptides. These composite structures combine the endosomal escape capability of the ELD with the cell penetration capability of the CPD, achieving both high transduction efficiency and reliable cytosolic delivery of polypeptide cargos.
2Productivity
If viral transduction methods are used for ex vivo manipulations, then cell transduction is achieved, but safety concerns and regulatory hurdles increase
Solution Approach 1:
The invention employs non-viral, peptide-based shuttle agents that are transient and do not integrate into the genome. These disposable-like delivery vectors provide the necessary transduction efficiency without the long-term safety concerns and regulatory burdens associated with viral vectors, enabling safer ex vivo manipulations.
Solution Approach 2:
The peptide-based shuttle agents serve as intermediary carriers that temporarily facilitate polypeptide cargo delivery into cells. Unlike viral vectors that permanently modify cells, these peptide mediators perform their delivery function and are then degraded, providing a safe alternative that maintains transduction efficiency without introducing persistent safety risks.
3Ease of operation
If polypeptide cargos are delivered using conventional methods, then some intracellular delivery is achieved, but endosomal trapping prevents proper subcellular localization
Solution Approach 1:
The invention ensures continuous useful action by designing the ELD-CPD shuttle agents to sequentially perform cell penetration followed by endosomal escape. This continuous process prevents cargo stagnation in endosomes and ensures reliable delivery to the cytosol, achieving both ease of intracellular delivery and precise subcellular localization.
Solution Approach 2:
The ELD component responds to pH parameter changes within the endosome to trigger membrane disruption and cargo release. This parameter-based activation mechanism ensures that cargo delivery is precisely controlled and occurs at the correct subcellular location, resolving the contradiction between ease of delivery and localization accuracy.
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 described peptides significantly increase the proportion of cells that can be transduced, facilitating the escape of endosomally-trapped cargos to the cytosol and enabling targeted subcellular delivery, thereby improving the efficiency and safety of polypeptide cargo delivery.
Implementation Method 1
synthetic peptides comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD, for use in increasing the transduction efficiency of an independent polypeptide cargo to the cytosol of a target eukaryotic cell
Implementation Method 2
the CPD enables intracellular delivery of said synthetic peptide
Data Source
AI summary
The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.