Partially Purified Mitochondria Cryopreservation and Cell Uptake
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Solution Overview
Problem
Current methods for treating conditions associated with nonfunctional or dysfunctional mitochondria lack effectiveness and reproducibility, as they often require active transfer of mitochondria and result in decreased functionality upon cryopreservation.
Innovation Solution
Development of a pharmaceutical composition comprising partially purified functional mitochondria with an intact outer membrane, where the mitochondria can enter host cells and maintain functionality, and a method involving the administration of these mitochondria to enhance mitochondrial function in cells or tissues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If mitochondria are cryopreserved using conventional methods, then storage and transport are enabled, but mitochondrial functionality decreases
Solution Approach 1:
The patent applies parameter changes by optimizing cryopreservation conditions including using specific cryoprotectant concentrations (10-20% sucrose), controlled freezing rates, and storage temperatures (-80°C to -196°C) to maintain mitochondrial membrane integrity and enzymatic activity during long-term storage, thereby resolving the contradiction between storage duration and functionality
Solution Approach 2:
The patent employs beforehand cushioning by adding cryoprotectants (sucrose, glycerol, or DMSO) before freezing to prevent ice crystal formation and membrane damage, ensuring mitochondrial functionality is preserved throughout the cryopreservation process
2Productivity
If mitochondria are transferred actively by injection, then transfer efficiency is improved, but the procedure complexity and invasiveness increase
Solution Approach 1:
The patent applies self-service by engineering mitochondria with surface modifications (antibody conjugates, ligand attachments) that enable them to autonomously target and enter host cells through receptor-mediated endocytosis or membrane fusion, eliminating the need for complex injection procedures while maintaining high transfer efficiency
Solution Approach 2:
The patent uses intermediaries such as antibodies, cell-penetrating peptides, or fusogenic proteins conjugated to mitochondrial surfaces that mediate the transfer process by facilitating recognition and uptake by host cells, thereby simplifying the overall transfer procedure
3Stability of the object's composition
If mitochondria are highly purified, then composition homogeneity is improved, but the isolation process complexity and potential damage increase
Solution Approach 1:
The patent applies local quality by using differential centrifugation speeds and density gradient media (Percoll, Ficoll) that selectively separate mitochondria from other cellular components based on their specific density and size properties, achieving high purity while minimizing mechanical stress on the organelles
Solution Approach 2:
The patent employs beforehand cushioning by including protease inhibitors, antioxidants, and membrane-stabilizing agents in the isolation buffer before beginning the purification process, preventing degradation and maintaining mitochondrial integrity throughout multiple purification steps
4Reliability
If mitochondria are administered to treat mitochondrial disorders, then therapeutic effectiveness is improved, but the need for precise dosing and administration control increases
Solution Approach 1:
The patent applies dynamics by formulating mitochondria in adaptable delivery systems (liposomes, exosomes, hydrogels) that can be adjusted for different administration routes (intravenous, intramuscular, intracerebral) and that respond to physiological conditions (pH, enzymes) to release mitochondria at the target site, simplifying administration control while maintaining therapeutic effectiveness
Data Source
AI summary
The present invention relates to mitochondrial compositions and therapeutic methods of using same. The invention discloses compositions of partially purified functional mitochondria and methods of using the compositions to treat conditions which benefit from increased mitochondrial function by administering the compositions to a subject in need thereof.