Exosome-ASO Constructs for CEBPβ Targeting

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Solution Overview

Problem

There is a need to improve the stability and targeting of antisense oligonucleotides (ASOs) for in vivo use, particularly in extracellular vesicles (EVs) like exosomes, to effectively deliver therapeutic agents that reduce gene expression associated with diseases such as cancer, as existing methods face challenges in immunogenicity and delivery efficiency.

Innovation Solution

Development of extracellular vesicles loaded with antisense oligonucleotides (ASOs) that are complementary to specific nucleic acid sequences within the CEBP/β transcript, incorporating scaffold proteins and modified nucleosides, which can target immune cells and reduce CEBP/β protein expression, using a contiguous nucleotide sequence of 10 to 30 nucleotides in length, and include sugar-modified nucleosides and phosphorothioate linkages for enhanced stability and specificity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional drug delivery methods are used to deliver ASOs, then delivery efficiency is limited, but immunogenicity is reduced

Engineering Contradiction:

Improvedelivery efficiencyVSAvoidimmunogenicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

Extracellular vesicles (EVs) are used as intermediary carriers to deliver ASOs into target cells. The EVs naturally fuse with cell membranes or are endocytosed, enabling efficient intracellular delivery of ASOs while their natural origin (e.g., exosomes from human cells) minimizes immunogenic responses. This resolves the contradiction by providing a biocompatible delivery vehicle that achieves both high delivery efficiency and low immunogenicity.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention creates a composite delivery system combining ASOs (nucleic acid therapeutics) with extracellular vesicles (biological carriers). This composite structure leverages the therapeutic functionality of ASOs while utilizing the EVs' natural cell-penetrating abilities and biocompatibility, achieving efficient delivery without triggering strong immune responses that would occur with synthetic delivery vehicles.

Inventive Principle:

Principle #40Composite materials

2Stability of the object's composition

If ASO sequence length is increased to improve binding affinity, then stability improves, but delivery efficiency decreases

Engineering Contradiction:

ImproveASO stabilityVSAvoiddelivery efficiency

Core Design Contradiction:

Stability of the object's compositionVSReliability

Solution Approach 1:

The extracellular vesicle membrane acts as a flexible protective shell that encapsulates ASOs of various lengths. This natural membrane structure protects even longer ASO sequences from degradation and steric hindrance during delivery, allowing stable, high-affinity ASOs to be delivered efficiently without the trade-off that would exist in conventional delivery systems.

Inventive Principle:

Principle #30Flexible shells and thin films

3Stability of the object's composition

If modified nucleosides and phosphorothioate linkages are incorporated to enhance stability, then ASO stability improves, but manufacturing complexity increases

Engineering Contradiction:

ImproveASO stabilityVSAvoidmanufacturing complexity

Core Design Contradiction:

Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The extracellular vesicle production system naturally incorporates modified nucleosides and phosphorothioate linkages when ASOs are introduced into producer cells. The cellular machinery processes and integrates these modifications during ASO incorporation into EVs, reducing the need for complex post-synthesis modification steps and simplifying the overall manufacturing process while maintaining high ASO stability.

Inventive Principle:

Principle #25Self-service

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 ASO-loaded EVs effectively reduce CEBP/β protein and mRNA levels in human immune cells by up to 100%, demonstrating improved stability and targeting capabilities, potentially offering a new therapeutic approach for cancer and other diseases by modulating gene expression.

Implementation Method 1

the ASO comprises a contiguous nucleotide sequence of 10 to 30 nucleotides in length that is complementary to a nucleic acid sequence within a CEBP/β transcript

Methodology Applied

Scientific EffectBase pairing: Chemical Bonding

Implementation Method 2

Exosomes comprise a membrane that encloses an internal space (i.e., lumen)

Methodology Applied

Scientific EffectPhysical containment: Physical Containment

Data Source

PatentUS20230002764A1Extracellular vesicle-ASO constructs targeting CEBP/beta

Publication Date: 2023.01.05 LONZA SALES AG

AI summary

The present disclosure relates to extracellular vesicles, e.g., exosomes, comprising an antisense oligonucleotide (ASO), wherein the ASO comprises a contiguous nucleotide sequence of 10 to 30 nucleotides in length that is complementary to a nucleic acid sequence within a CEBP/transcript. Also provided herein are methods for producing the exosomes and methods for using the exosomes to treat and/or prevent diseases or disorders.