Dual-Protease Cleavable Polypeptides for Disease-Site Activation

The patent applies local quality by designing substrates with specific amino acid sequences (such as HQSRS for MMPs or AIALYAD for serine proteases) that are selectively recognized by dysregulated proteases in disease tissues. The substrate sequences are optimized to have high affinity for specific proteases while maintaining resistance to other proteases, thereby achieving localized activation only at disease sites and minimizing off-target effects in healthy tissues.

The patent utilizes parameter changes by modifying the amino acid sequence parameters of the substrate to optimize protease recognition. Specific sequences like HQSRS with particular amino acid compositions are designed to maximize binding affinity to target proteases while minimizing cross-reactivity. The cleavable moiety sequences are carefully selected to ensure efficient cleavage by disease-associated proteases but resistance to normal physiological proteases.

The patent applies segmentation by dividing the activation function into separate cleavable moieties, each targeting a specific protease pathway. The polypeptide contains distinct functional segments such as an MMP-targeting moiety (e.g., HQSRS) and a serine protease-targeting moiety (e.g., AIALYAD), which can be independently optimized and combined. This modular approach allows simultaneous targeting of multiple protease families while maintaining manageable structural complexity through standardized linkage regions.

The patent implements multi-functionality by creating a single polypeptide that performs multiple activation functions through different cleavable moieties. The same polypeptide can be activated by both MMPs and serine proteases depending on the disease context, allowing one molecule to address multiple protease-driven pathologies. The universal design framework uses consistent structural elements (such as the P1-P4 cleavage site architecture) across different protease targets to manage complexity.