Matriptase and uPA-Cleavable Antibody Substrates for Targeted Activation
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Solution Overview
Problem
There is a need to identify new substrates for proteases such as matriptase and u-plasminogen activator (uPA) for therapeutic, diagnostic, and prophylactic indications, as deregulated expression and activity of these enzymes are associated with various pathological conditions.
Innovation Solution
Development of isolated polypeptides comprising a cleavable moiety (CM) that serves as a substrate for matriptase or uPA, linked to an antibody or its antigen-binding fragment (AB) via a masking moiety (MM), which inhibits binding to the antigen until cleaved by the protease, allowing release of linked agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a masking moiety is used to inhibit antibody binding to the antigen, then the antibody activity is suppressed in the uncleaved state, but the antibody cannot bind the target until protease cleavage occurs
Solution Approach 1:
The antibody is divided into functional segments: an antigen-binding fragment (AB) and a masking moiety (MM) connected by a cleavable moiety (CM). The MM contains a protease-specific substrate sequence that, when cleaved by the target protease, releases the AB to bind the antigen. This segmentation enables controlled activation while maintaining overall molecular integrity.
Solution Approach 2:
The cleavable moiety acts as an intermediary between the masking moiety and the antigen-binding fragment. It contains a peptide sequence recognized by the target protease, serving as a molecular switch that transitions the antibody from an inactive masked state to an active unmasked state upon protease-mediated cleavage.
2Adaptability or versatility
If new substrates for proteases are identified and used, then therapeutic and diagnostic applications are enabled, but the need to identify and characterize new substrate sequences increases research complexity
Solution Approach 1:
The patent provides a universal framework for designing protease-activatable antibodies by identifying conserved substrate sequence motifs for specific proteases (e.g., MMP-2, MMP-9, uPA). These standardized substrate sequences can be incorporated into different antibody constructs targeting various antigens, enabling broad application across multiple therapeutic and diagnostic indications without requiring de novo substrate identification for each application.
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
Enables targeted delivery and activation of therapeutic or diagnostic agents by protease-specific cleavage, enhancing therapeutic efficacy and diagnostic accuracy.
Implementation Method 1
a masking moiety (MM) that inhibits the ability of the AB to bind the antigen
Implementation Method 2
Proteases are enzymes that cleave the peptide bonds between amino acid residues
Data Source
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AI summary
The invention relates generally to polypeptides that include a cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator (uPA), to activatable antibodies and other larger molecules that include the cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator, and to methods of making and using these polypeptides that include a cleavable moiety that is a substrate for at least one protease selected from matriptase and uplasminogen activator in a variety of therapeutic, diagnostic and prophylactic indications.