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Updated: Jun 24, 2026
Lighting Up the Pathways to Caspase Activation Using Bimolecular Fluorescence Complementation
Published on: March 5, 2018
Establishment of a high-throughput screening system for caspase-3 inhibitors
Abstract:
In most tissues, apoptosis plays a pivotal role in normal development and for regulating cell number, thus inappropriate apoptosis underlies a variety of diseases. Caspase-3 is one of a family of caspases that are mainly involved in the apoptotic signal transduction pathway, where caspase-3 acts as an effect molecule to proteolytically cleave intracellular substrates that are necessary for maintaining cell survival. Recent evidences show that apoptotic cell death can be blocked by inhibiting caspase-3, suggesting its inhibitors have potential to be therapeutic drugs for the diseases related with inappropriate apoptosis. We have established a screening system to search caspase-3 inhibitors from chemical libraries stocked in our institute. The enzyme assay is configured entirely in 96-well format, which is easily adapted for high throughput screening. Before performing mass screening, 80 in-house compounds were screened as a preliminary experiment, and we found that morin hydrate inhibited caspase-3 by 66.4% at the final concentration of 20 microM.
Insights
Researchers screened for caspase-3 inhibitors, crucial for controlling cell death and related diseases. Morin hydrate emerged as a promising compound, inhibiting caspase-3 activity by 66.4% in preliminary tests.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Apoptosis, or programmed cell death, is vital for development and tissue homeostasis.
- Dysregulated apoptosis is implicated in various human diseases.
- Caspase-3 is a key executioner enzyme in the apoptotic pathway, making it a therapeutic target.
Purpose of the Study:
- To establish a high-throughput screening system for identifying caspase-3 inhibitors.
- To discover novel chemical compounds that can inhibit caspase-3 activity.
- To explore potential therapeutic agents for diseases associated with inappropriate apoptosis.
Main Methods:
- Development of a 96-well format enzyme assay for caspase-3 inhibition screening.
- Screening of 80 in-house compounds using the established assay.
- Quantification of caspase-3 inhibition by candidate compounds.
Main Results:
- A robust, high-throughput screening system for caspase-3 inhibitors was successfully established.
- Morin hydrate demonstrated significant inhibition of caspase-3 activity.
- Morin hydrate inhibited caspase-3 by 66.4% at a concentration of 20 microM in preliminary screening.
Conclusions:
- Inhibitors of caspase-3 hold therapeutic potential for diseases linked to aberrant apoptosis.
- Morin hydrate is a promising lead compound for further investigation as a caspase-3 inhibitor.
- The developed screening platform is suitable for large-scale identification of novel caspase-3 inhibitors.
