Fusion Polypeptide Cargo Delivery With Non-Covalent Cell Penetration

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Solution Overview

Problem

Existing cell-penetrating peptides (CPPs) are inefficient for delivering a wide variety of cargoes across different cell types and organisms, require high molecular weights, and are difficult to produce in sufficient quantities, leading to low uptake efficiency and limited applications.

Innovation Solution

A fusion polypeptide comprising a substrate binding domain, a cell penetrating domain, a signal sequence, and optionally a pH-sensitive autoprotease domain, designed to efficiently deliver a wide range of substrates into cells, including a substrate binding domain, a cell penetrating domain, and a substrate binding domain, a cell penetrating domain, and a signal sequence, which can be produced in recombinant systems, and is capable of non-covalently binding to various cargoes such as polypeptides, polysaccharides, DNA, and RNA, facilitating their delivery through endocytosis or non-endosomal mechanisms.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If cell-penetrating peptides (CPPs) are used to deliver cargo into cells, then cargo delivery is enabled, but uptake efficiency is low and high molecular weights are required

Engineering Contradiction:

Improvecargo delivery efficiencyVSAvoidmolecular weight of CPP

Core Design Contradiction:

ProductivityVSWeight of moving object

Solution Approach 1:

The fusion polypeptide is divided into distinct functional domains: a substrate binding domain (50-200 amino acids) and a cell penetrating domain (15-50 amino acids). This segmentation allows each domain to perform its specific function optimally while keeping the overall molecular weight lower than conventional CPPs, resolving the contradiction between delivery efficiency and molecular weight requirements.

Inventive Principle:

Principle #1Segmentation

2Productivity

If high amounts of CPP are used to achieve sufficient cargo delivery, then cargo delivery is improved, but production difficulty increases

Engineering Contradiction:

Improvecargo delivery efficiencyVSAvoidproduction ease of CPP

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The substrate binding domain is designed with universal binding capabilities that can accommodate various cargo types (polypeptides, polysaccharides, DNA, RNA) through non-covalent interactions. This multi-functionality allows a single fusion polypeptide design to deliver diverse cargoes efficiently, reducing the need for producing multiple different CPP variants and simplifying manufacturing processes.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Productivity

If CPPs are used for cargo delivery, then cargo transport into cells is enabled, but cargo stability is compromised due to covalent linkage requirements

Engineering Contradiction:

Improvecargo transport efficiencyVSAvoidcargo stability

Core Design Contradiction:

ProductivityVSStability of the object's composition

Solution Approach 1:

The substrate binding domain acts as an intermediary between the cell penetrating domain and the cargo. It binds cargo through non-covalent interactions (hydrogen bonds, hydrophobic interactions, electrostatic interactions), which are sufficient for stable transport but allow the cargo to maintain its natural structure and stability, unlike covalent linkages that would compromise cargo integrity.

Inventive Principle:

Principle #24Intermediary (Mediator)

4Reliability

If CPPs are designed for specific cell types or organisms, then targeted delivery is improved, but adaptability to different cell types and organisms decreases

Engineering Contradiction:

Improvetargeted delivery reliabilityVSAvoidadaptability to different cell types and organisms

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The fusion polypeptide design allows for parameter changes in the substrate binding domain to adapt to different cargo types and target cell types. By modifying the binding domain's amino acid sequence while maintaining the core cell penetrating domain, the system can be optimized for specific applications (e.g., plant cells vs. mammalian cells) without losing overall versatility, thus resolving the contradiction between targeted delivery reliability and adaptability.

Inventive Principle:

Principle #35Parameter changes

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 fusion polypeptide achieves high efficiency in delivering diverse cargoes across different cell types and organisms, maintaining cargo stability and enabling targeted delivery, even in plant cells, with the potential for economic production and scalability.

Implementation Method 1

a substrate binding domain binding to a substrate non-covalently, wherein the substrate is selected from the group consisting of polypeptides, polysaccharides, DNA, RNA or a mixture of DNA and RNA

Methodology Applied

Scientific EffectNon-covalent binding: Adsorption

Implementation Method 2

CPPs are assumed to enable the membrane passing of fusion polypeptides by non-endosomal mechanisms

Methodology Applied

Scientific EffectEndocytosis:

Data Source

PatentUS20260009040A1Fusion polypeptides for cell penetration and cell targeting

Publication Date: 2026.01.08 KUTZNER CHRISTOPH

AI summary

The present invention concerns fusion polypeptides comprising a cell penetration and a substrate binding domain. Additionally, the present invention concerns a genetically modified cell expressing these fusion polypeptides as well as the use of these fusion polypeptides for introducing a substance into a target cell.