Stable Exosome Formulations via Lyophilization

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Solution Overview

Problem

Current exosome formulations for clinical use lack stability, particularly in maintaining the levels of miR-210 and miR-146A, which are crucial for their therapeutic benefits in cardiac tissue repair, and face challenges in long-term storage and bioactivity retention.

Innovation Solution

Development of stable lyophilized and liquid exosome formulations generated from cardiosphere-derived cells using ultrafiltration and diafiltration, which maintain at least 90% of miR-210 and miR-146A levels and in vitro/in vivo biological activity for extended periods, including after lyophilization and rehydration.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Duration of action of stationary object

If exosomes are stored in liquid form at refrigerated or frozen temperatures, then their storage duration is extended, but their stability and bioactivity are compromised over time

Engineering Contradiction:
Improvestorage durationVSAvoidbioactivity retention
Core Design Contradiction:
Duration of action of stationary objectVSReliability

Solution Approach 1:

The patent applies freeze-drying (lyophilization) technology to transform exosomes from liquid state to solid dried state, and then rehydrates them back to liquid form for storage. This phase transition process removes water completely during freezing and drying, then restores it controllably, achieving both extended storage duration and maintained bioactivity without requiring continuous refrigeration or freezing.

Inventive Principle:
Principle #36Phase transitions

Solution Approach 2:

The invention changes the physical state parameters of exosomes from liquid to dried powder form through controlled freeze-drying processes. By adjusting parameters such as freezing temperature, drying time, and rehydration conditions, the patent achieves stable storage at ambient temperatures while preserving miR-210 and miR-146A levels and biological activity.

Inventive Principle:
Principle #35Parameter changes

2Stability of the object's composition

If exosomes are lyophilized and rehydrated, then storage stability is improved, but manufacturing complexity increases

Engineering Contradiction:
Improveformulation stabilityVSAvoidmanufacturing process complexity
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent performs preliminary freeze-drying of exosome formulations before final storage preparation. By pre-freezing and lyophilizing the exosomes in controlled manufacturing conditions, the formulation is stabilized in advance, allowing simple storage and transport without complex temperature control systems during distribution and use.

Inventive Principle:
Principle #10Preliminary action

3Quantity of substance

If conventional isolation methods are used, then exosome yield is achieved, but miR-210 and miR-146A levels deteriorate during storage

Engineering Contradiction:
Improveexosome yieldVSAvoidmiR-210 and miR-146A degradation
Core Design Contradiction:
Quantity of substanceVSLoss of substance

Solution Approach 1:

The patent uses freeze-drying to remove water from exosome formulations, transforming them from liquid to solid state. This phase transition prevents degradation of miR-210 and miR-146A by eliminating the aqueous environment that facilitates enzymatic degradation and chemical breakdown, thereby preserving these critical molecules during long-term storage.

Inventive Principle:
Principle #36Phase transitions

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 stable formulations retain high levels of miR-210 and miR-146A, along with bioactivity, even after storage and lyophilization, demonstrating improved stability and effectiveness in cardioprotective and angiogenic functions.

Implementation Method 1

stable liquid cardiosphere-derived cell (CDC) exosome formulation suitable for administration to a human, wherein the exosome formulation is generated from CDCs by ultrafiltration and diafiltration

Methodology Applied
Scientific EffectUltrafiltration: Semipermeable Membrane

Implementation Method 2

stable liquid cardiosphere-derived cell (CDC) exosome formulation suitable for administration to a human, wherein the exosome formulation is generated from CDCs by ultrafiltration and diafiltration

Methodology Applied
Scientific EffectDiafiltration: Semipermeable Membrane

Implementation Method 3

stable lyophilized exosome formulation suitable for administration to a human comprising at least 10 6 particles/mL or 10 8 particles/mL

Methodology Applied
Scientific EffectFreeze-drying: Freeze Drying

Data Source

PatentEP3226875B1Processes for producing stable exosome formulations
Publication Date: 2020.05.27 CAPRICOR INC
  • EP3226875B1 patent drawingFigure 1A~1B
  • EP3226875B1 patent drawingFigure 1C~1D
  • EP3226875B1 patent drawingFigure 2

AI summary

The invention encompasses methods for generating stable exosome formulations and encompasses stable exosome formulations. The exosome formulations encompass stable liquid exosome formulations and stable lyophilized exosome formulations. In some embodiments, the exosome formulations can be generated by ultrafiltration and diafiltration. The exosome formulations can be suitable for administration to a human.