Protease-Cleavable Polypeptides for Selective Disease Tissue Activation
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Solution Overview
Problem
There is a need to identify new substrates for proteases that can be used in therapeutic, diagnostic, and prophylactic applications, as existing substrates are resistant to cleavage by specific proteases and lack efficient activation mechanisms.
Innovation Solution
Development of isolated polypeptides with specific cleavable moieties (CM1 and CM2) that are cleavable by proteases such as MMP2, MMP9, MMP14, MT-SP1, and uPA, with enhanced cleavability and in vivo stability, allowing activation of an active moiety (AM) for therapeutic, diagnostic, or prophylactic use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing substrates are used for protease targeting, then protease activity can be detected or modulated, but the substrates are not effectively targeted to dysregulated protease activity in disease tissues, resulting in lack of selectivity
Solution Approach 1:
The substrate is engineered with disease-specific peptide sequences that are recognized preferentially by proteases in diseased tissues. This local adaptation of substrate structure allows selective targeting to dysregulated protease activity in specific disease contexts while maintaining the ability to be applied across different disease types by changing the peptide sequence.
2Adaptability or versatility
If protease substrates are designed for broad protease recognition, then they can be applied to multiple disease types, but they lack selectivity for dysregulated protease activity in specific disease tissues
Solution Approach 1:
The substrate design utilizes variable peptide sequences where specific amino acid compositions and arrangements can be changed to match the substrate specificity profiles of different proteases. By adjusting these sequence parameters, the same substrate framework can be adapted to target different proteases in different disease contexts while maintaining high selectivity for each specific protease-disease pairing.
3Reliability
If new protease substrates are developed for specific disease targeting, then selectivity and therapeutic index are improved, but the complexity of substrate design and validation increases
Solution Approach 1:
The substrate is divided into distinct functional segments: a disease-specific peptide recognition sequence, a linker region, and a reporter or therapeutic payload. This segmentation allows independent optimization of each component - the peptide sequence for protease specificity, the linker for stability and cleavage efficiency, and the payload for therapeutic effect - thereby reducing overall design complexity while improving therapeutic index.
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 polypeptides demonstrate high cleavability and in vivo stability, enabling efficient activation of AMs with enhanced masking efficiency and cleavability, suitable for therapeutic, diagnostic, or prophylactic applications.
Implementation Method 1
Proteases are enzymes that catalyze the hydrolysis of peptide bonds between amino acid residues
Implementation Method 2
Proteases are enzymes that catalyze the hydrolysis of peptide bonds between amino acid residues
Data Source
AI summary
Isolated polypeptides that include a cleavable moiety that is a substrate for at least one protease (e.g., MT-SP1 and/or an MMP) and isolated polypeptides that include a substrate that has a first cleavable moiety cleavable by a first protease and a second cleavable moiety cleavable by a second protease are disclosed. Activatable molecules including the isolated polypeptides are disclosed. Methods of making and using the isolated polypeptides and activatable molecules including the isolated polypeptides in a variety of therapeutic, diagnostic, and prophylactic applications are disclosed.