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Prodrug-inspired probes selective to cathepsin B over other cysteine cathepsins
Morshed A Chowdhury1, Ignace A Moya, Shardul Bhilocha
1Thunder Bay Regional Research Institute, 2321-290 Munro Street, Thunder Bay, Ontario, Canada , P7A 7T1.
Abstract:
Cathepsin B (CTB) is a cysteine protease believed to be an important therapeutic target or biomarker for several diseases including aggressive cancer, arthritis, and parasitic infections. The development of probes capable of assessing CTB activity in cell lysates, living cells, and animal models of disease are needed to understand its role in disease progression. However, discovering probes selective to cathepsin B over other cysteine cathepsins is a significant challenge due to overlap of preferred substrates and binding site homology in this family of proteases. Herein we report the synthesis and detailed evaluation of two prodrug-inspired fluorogenic peptides designed to be efficient and selective substrate-based probes for CTB. Through cell lysate and cell assays, a promising lead candidate was identified that is efficiently processed and has high specificity for CTB over other cysteine cathepsins. This work represents a key step toward the design of rapid release prodrugs or substrate-based molecular imaging probes specific to CTB.
Insights
Researchers developed selective fluorogenic peptide probes to measure Cathepsin B (CTB) activity. This breakthrough aids in understanding diseases like cancer and developing targeted therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Cathepsin B (CTB) is a cysteine protease implicated in aggressive cancers, arthritis, and parasitic infections.
- Accurate assessment of CTB activity is crucial for disease understanding and therapeutic development.
- Developing selective probes for CTB over other cysteine cathepsins presents a significant challenge due to conserved enzymatic properties.
Purpose of the Study:
- To design and synthesize novel, selective, substrate-based fluorogenic peptide probes for Cathepsin B (CTB).
- To evaluate the efficiency and specificity of these probes in various biological contexts, including cell lysates and living cells.
Main Methods:
- Synthesis of two prodrug-inspired fluorogenic peptide substrates.
- In vitro evaluation using cell lysates to assess probe processing and specificity.
- Cell-based assays to determine probe performance in a cellular environment.
Main Results:
- One lead probe candidate demonstrated efficient processing and high specificity for CTB.
- The probe effectively distinguished CTB activity from that of other cysteine cathepsins.
- Successful identification of a probe suitable for monitoring CTB activity.
Conclusions:
- The developed fluorogenic peptide probe is a promising tool for assessing Cathepsin B activity.
- This work facilitates the design of targeted prodrugs and molecular imaging agents for CTB.
- The findings represent a significant advancement in the field of protease-based diagnostics and therapeutics.
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