Binary Enzyme System for Selective Tumor Cell Apoptosis
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Solution Overview
Problem
Current cancer treatments, such as chemotherapy and targeted therapies, often harm fast-dividing normal cells alongside cancer cells, leading to significant side effects and an immunological burden, while gene therapy and genetic vaccination methods face limitations in specificity and efficiency in inducing cell death in tumor cells.
Innovation Solution
A binary enzyme system comprising an inactive form of granzyme B and tobacco etch virus protease is introduced into tumor cells, where the protease recognizes and cleaves a specific sequence to activate granzyme B, inducing apoptosis in cancer cells through a specific and efficient mechanism.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chemotherapy is used to kill tumor cells, then cell division disruption is achieved, but normal rapidly dividing cells are also damaged causing side effects
Solution Approach 1:
The invention divides the therapeutic system into two separate components: an inactive proenzyme (granzyme B precursor) and an activating enzyme (TEV protease). The proenzyme is delivered to tumor cells but remains inactive until specifically activated by TEV protease, which is also delivered to the same cells. This segmentation allows the therapeutic agent to be present in the target cells without immediately exerting its cytotoxic effect, thereby avoiding damage to normal cells while maintaining the ability to selectively induce apoptosis in tumor cells when activated.
Solution Approach 2:
The invention introduces TEV protease as an intermediary activating agent. The proenzyme is designed with a specific recognition sequence that TEV protease can cleave. This intermediary mechanism ensures that the active form of granzyme B is only generated within tumor cells that have received both the proenzyme and the activating protease, providing selective activation and minimizing off-target effects on normal cells.
2Measurement precision
If targeted therapies are used to eliminate cancer cells, then specificity is improved, but immunological burden increases and treatment frequency is limited
Solution Approach 1:
The invention employs a disposable binary enzyme system where the proenzyme and activating protease are delivered as separate entities that act locally within tumor cells. The system does not rely on long-term immune system activation or regeneration, but rather on direct enzymatic action that can be repeated without imposing cumulative immunological burden. The enzymes perform their function and are metabolized, allowing for more frequent treatments compared to immunotherapies that require immune system recovery time.
3Productivity
If granzyme B is activated in tumor cells, then apoptosis is induced efficiently, but non-specific activation would harm normal cells
Solution Approach 1:
The invention implements preliminary action by delivering the inactive proenzyme to tumor cells in advance, where it accumulates but remains dormant. The proenzyme is designed with a built-in recognition sequence that awaits the activating protease. This preliminary positioning of the inactive form ensures high concentration in target cells without immediate cytotoxic effect, and activation only occurs when the specific protease is present, preventing premature or non-specific activation that would damage normal cells.
Solution Approach 2:
The invention applies local quality by creating a unique microenvironment within tumor cells that contains both the proenzyme and the activating TEV protease. The proenzyme is engineered with a specific recognition sequence (ENLYFQ) that only TEV protease can cleave. This localized specificity ensures that granzyme B activation occurs only in cells that have received both components, creating a locally controlled therapeutic effect that spares normal cells lacking the activating protease or the specific recognition sequence.
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 allows for targeted and efficient induction of cell death in tumor cells, minimizing harm to normal cells and reducing the immunological burden, with the potential for improved therapeutic outcomes in cancer treatment.
Implementation Method 1
TEV recognizes the recognition site (recognition sequence) ENLYFQ (SEQ ID No. 6) or ENLYFQG (SEQ ID No. 7) in the inactive form, which is linked (ligated) to the N-terminus of SEQ ID No. 2
Implementation Method 2
TEV (SEQ ID No. 4 or SEQ ID No. 5) or a coding nucleic acid thereof or a functional variant thereof, wherein SEQ ID No. 2 is obtained or released by cleavage by TEV (SEQ ID No. 4 or SEQ ID No. 5)
Implementation Method 3
Once the inactive form of Granzyme B and TEV are introduced into a tumor cell, either together or separately, and possibly expressed, TEV releases the active form of Granzyme B according to SEQ ID No. 2, and consequently, induced cell death occurs via apoptosis or programmed cell death
Implementation Method 4
These mechanisms and proteins have in common is that they activate a proteolytic cellular cascade series of cysteinyl proteases, called caspases. The initially activated caspases, such as caspase 8 and caspase 9, then activate effector cascades, such as caspases 3 and 6.
Data Source
AI summary
The invention relates to a combination preparation containing a selective cell-death-inducing binary enzyme system for use in the therapy and/or treatment of cancer and tumours in humans and animals, a method and the use thereof.