Split-Intein Polypeptide Composition for Selective Tumor Toxicity
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Solution Overview
Problem
Immunotoxins exhibit non-specific toxicity to normal tissues due to the expression of tumor-associated antigens in both malignant tumors and normal cells, posing safety concerns and limiting their therapeutic efficacy.
Innovation Solution
A polypeptide composition comprising a first polypeptide with a first toxin fragment and a first intein fragment, and a second polypeptide with a second toxin fragment and a second intein fragment, where the toxin fragments are non-biotoxic and interact to form a biotoxic toxin through intein-mediated splicing, allowing targeted delivery to tumor cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If immunotoxin is used to target and kill tumor cells, then the specific killing effect is improved, but non-specific toxicity to normal tissue cells occurs
Solution Approach 1:
The toxin is divided into two separate fragments (first toxin fragment and second toxin fragment) that are individually non-biotoxic. These fragments are fused to split intein segments and expressed separately. Only when both fragments come together through intein trans-splicing do they form the complete biotoxic toxin, thereby achieving segmentation of the toxic function to reduce non-specific toxicity while maintaining specific killing effect.
Solution Approach 2:
The two toxin fragments are pre-expressed in fusion with split intein segments in host cells. The intein-mediated trans-splicing is triggered under specific conditions (such as oxidative environment) to assemble the complete toxin only when needed and at the target site, rather than having the complete toxin present from the beginning. This preliminary expression of inactive fragments followed by conditional activation reduces off-target effects.
2Object-affected harmful factors
If the toxin is split into two inactive fragments for separate expression, then the safety and non-specific toxicity are reduced, but the device complexity and manufacturing difficulty increase
Solution Approach 1:
The split intein system acts as an intermediary mechanism that facilitates the controlled assembly of the two toxin fragments. The intein segments are fused to the toxin fragments and mediate their specific pairing and splicing through a well-defined biochemical mechanism (intein trans-splicing). This intermediary system provides a reliable and specific method to reconstitute the active toxin only from the correct fragment pairing, reducing the need for complex external assembly protocols.
3Reliability
If the first polypeptide and second polypeptide are mixed to form the biotoxic toxin, then the therapeutic efficacy is enhanced, but the loss of time for assembly and activation occurs
Solution Approach 1:
The two toxin fragments are pre-expressed and prepared in advance in fusion with split intein segments. The fragments are synthesized and stabilized in their inactive state, ready for rapid assembly. When triggered by the appropriate stimulus (such as oxidative conditions in the tumor microenvironment), the intein-mediated splicing occurs quickly to form the active toxin, minimizing the time loss between administration and activation.
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 composition achieves selective toxicity to tumor cells by forming a biotoxic toxin only upon interaction, reducing off-target effects and enhancing therapeutic efficacy while minimizing harm to normal tissues.
Implementation Method 1
the first polypeptide and the second polypeptide may make the first toxin fragment and the second toxin fragment to form a biotoxic toxin by means of the interaction between the first intein fragment and the second intein fragment
Data Source
Figure 1A~2C
Figure 3~4
Figure 5~6
AI summary
Provided is a polypeptide composition, comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprise a first toxin fragment and a first intein fragment, the second polypeptide comprise a second toxin fragment and a second intein fragment, the first polypeptide is different from the second polypeptide; the first toxin fragment and the second toxin fragment are non-biotoxic; the first polypeptide and the second polypeptide may make the first toxin fragment and the second toxin fragment into biotoxic toxins by means of the interaction of the first intein fragment and the second intein fragment. Also provided a method of preparing the composition, a kit comprising the first polypeptide and the second polypeptide, and the use and method of the composition and kit in treating tumors.