Establishment of a high-throughput screening system for caspase-3 inhibitors.
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.
Park
S Y
SY
Department of Veterinary Medicine, Konkuk University, Seoul, Korea.
Park
S H
SH
Lee
I S
IS
Kong
J Y
JY
D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D061945:Caspase Inhibitors; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli; D006801:Humans
D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; C505407:CASP3 protein, human; D053148:Caspase 3