Reversible Thrombin Stabilization via Oligonucleotide Binding
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Solution Overview
Problem
Current methods for stabilizing thrombin in liquid form are inadequate, as they either require expensive freezing, non-specific protease inhibitors that can affect other proteases, or result in lyophilized preparations that are costly and time-consuming to reconstitute, and existing stabilizing compounds either inactivate thrombin or require higher concentrations.
Innovation Solution
The use of reversible thrombin binding oligonucleotides that can inhibit thrombin activity in an aqueous solution, allowing for stabilization without freezing or reconstitution, and can be reversed with an antisense oligonucleotide to restore thrombin activity when needed.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If liquid thrombin is stored without freezing or protease inhibitors, then ease of operation and clinical usability are improved, but thrombin activity is lost due to autolysis during storage
Solution Approach 1:
A reversible thrombin inhibitor is introduced as an intermediary substance that binds to thrombin during storage to prevent autolysis, and can be removed before clinical use to restore thrombin activity. This mediator allows the thrombin to remain stable in liquid form without freezing or complex protease inhibitor formulations, solving both the stability and usability problems.
2Reliability
If protease inhibitors are added to stabilize thrombin, then thrombin activity is preserved during storage, but other proteases in the hemostasis pathway are adversely affected
Solution Approach 1:
The reversible thrombin inhibitor is designed to be highly specific to thrombin, affecting only the target protein during storage. The inhibitor does not interact with other proteases in the hemostasis pathway, providing localized stabilization without the promiscuous effects of conventional protease inhibitors.
3Ease of operation
If thrombin is formulated at low concentration to reduce viscosity, then ease of administration is improved, but larger amounts must be administered achieving the same therapeutic effect
Solution Approach 1:
The reversible thrombin inhibitor is added during storage to prevent autolysis, allowing thrombin to be formulated at higher concentrations without loss of activity. This preliminary stabilization enables the thrombin to maintain its potency, so that smaller volumes can be administered to achieve the desired therapeutic effect.
4Duration of action of stationary object
If lyophilization is used to stabilize thrombin, then shelf-life is extended, but manufacturing complexity and cost increase, and yield is reduced
Solution Approach 1:
The reversible thrombin inhibitor allows thrombin to be stored in liquid form at controlled temperatures without freezing, changing the storage parameters from extreme cold or dry conditions to moderate refrigeration. This eliminates the need for lyophilization while maintaining stability, simplifying manufacturing and reducing costs.
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
This approach effectively extends the shelf-life of thrombin, maintaining its activity and allowing for easy use in clinical settings without the need for freezing or reconstitution, while avoiding the drawbacks of existing methods.
Implementation Method 1
the thrombin binding oligonucleotides are such that they can bind to thrombin in a thrombin aqueous solution and inhibit, at least partially, the activity of thrombin
Implementation Method 2
the inhibition of thrombin activity can be reversed by contacting the thrombin solution comprising the thrombin binding oligonucleotide with an antisense oligonucleotide
Data Source
Figure 1
Figure 2A~2B
Figure 2C~2D
AI summary
The present invention is directed to compounds, methods for stabilizing thrombin activity with a thrombin binding oligonucleotide and to stabilized thrombin. The thrombin binding oligonucleotide is capable of inhibiting thrombin activity whereby the inhibition can be reversed with an antisense oligonucleotide.