Alphabody Polypeptides for Intracellular Delivery

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Macromolecules, such as polypeptides and nucleic acids, face difficulties in crossing cellular membranes to reach intracellular targets due to unfavorable biological and physicochemical properties, limiting their therapeutic and diagnostic applications.

Innovation Solution

Development of Alphabody polypeptides with a positively charged internalization region, comprising at least six positively charged amino acid residues, that can penetrate cell membranes and specifically bind to intracellular targets, enhancing their stability and functionality within cells.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If macromolecules are used as therapeutic agents to target intracellular components, then binding specificity and stability are improved, but the ability to cross cellular membranes deteriorates

Engineering Contradiction:

Improvebinding specificityVSAvoidcell membrane permeability

Core Design Contradiction:

ReliabilityVSEase of operation

Solution Approach 1:

The polypeptide is divided into distinct functional segments: an internalization region (with positively charged residues for membrane penetration) and a binding region (for specific target recognition). This segmentation allows each region to independently perform its specialized function, resolving the contradiction between membrane permeability and binding specificity

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

Different regions of the polypeptide are assigned different local properties: the internalization region contains clustered positively charged amino acids (lysine, arginine) for electrostatic interaction with membranes, while the binding region maintains the structural characteristics needed for specific target binding. This local differentiation enables simultaneous optimization of both membrane crossing and specific binding

Inventive Principle:

Principle #3Local quality

2Ease of operation

If delivery vehicles are used to facilitate internalization, then cellular uptake is improved, but accumulation in lysosomes and degradation deteriorates

Engineering Contradiction:

Improvecellular uptakeVSAvoidintracellular stability

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The internalization region acts as an intermediary that mediates membrane crossing through electrostatic interactions, but is designed to release the bound effector molecule after internalization. This allows the effector to be delivered into the cytoplasm without being trapped in endocytic pathways leading to lysosomal degradation

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The polypeptide uses specific parameter changes in charge distribution (clustered positive charges in internalization region vs. neutral/polar charges in binding region) and structural organization (amphipathic helices) to switch between membrane-binding mode and effector-release mode, ensuring delivery to the correct intracellular compartment while avoiding degradation pathways

Inventive Principle:

Principle #35Parameter changes

3Ease of operation

If polypeptides are designed with positively charged regions for internalization, then membrane penetration is improved, but structural stability may deteriorate

Engineering Contradiction:

Improvemembrane penetrationVSAvoidpolypeptide structural stability

Core Design Contradiction:

Ease of operationVSStability of the object's composition

Solution Approach 1:

The polypeptide combines different types of amino acid residues with complementary properties: positively charged residues (lysine, arginine) for membrane interaction, hydrophobic residues for structural core formation, and polar residues for binding specificity. This composite composition allows the structure to simultaneously achieve membrane penetration capability and structural stability

Inventive Principle:

Principle #40Composite materials

Solution Approach 2:

The internalization region forms amphipathic alpha-helical structures with specific curvature and geometry that facilitate membrane interaction while maintaining structural integrity. The helical configuration provides both the positive charge presentation needed for membrane binding and the structural stability required to resist degradation

Inventive Principle:

Principle #14Spheroidality (Curvature)

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 Alphabody polypeptides effectively enter cells and maintain stability, enabling targeted modulation of intracellular processes and addressing previously 'undruggable' protein targets, thereby expanding therapeutic options for various diseases.

Implementation Method 1

capable of being internalized into a cell through the presence of at least one positively charged internalization region

Methodology Applied

Scientific EffectElectrostatic interaction: Electrostatics

Data Source

PatentUS10023647B2Polypeptides capable of cellular internalization

Publication Date: 2018.07.17 COMPLIX NV

AI summary

Provided herein are polypeptides that are capable of crossing the cellular membrane and entering into the intracellular environment, which polypeptides are suitable for use in prophylactic, therapeutic or diagnostic applications as well as in screening and detection. Nucleic acids encoding such polypeptides; methods for preparing such polypeptides, host cells expressing or capable of expressing such polypeptides, compositions, and in particular pharmaceutical compositions, that comprise such polypeptides, in particular for prophylactic, therapeutic or diagnostic purposes are also provided.