BCI-121 is a substrate-competitive SMYD3 inhibitor that inhibits the proliferation of the cancer cell.

BCI-121 significantly reduced the proliferation of HT29 (by 46%) and HCT116 (by 54%) cells at 72 h and decreased the expression levels of SMYD3 target genes. BCI-121 treatment only affected proliferation of cancer cell lines expressing high levels of SMYD3. The high levels of SMYD3 expression detected in OVCAR-3 cells, along with the strong reduction in proliferation rate observed upon BCI-121 treatment.

Cell proliferation is determined using the cell proliferation reagent WST-1. Cells are seeded into 96-well plates one day before treatment. After 48 h, 72 h, or 96 h of BCI-121 or DMSO exposure, 10 μL of the Cell Proliferation Reagent WST-1 are added to each well and incubated at 37 °C in a humidified incubator for up to 1 h. Absorbance is measured on a microplate reader at 450/655 nm. The proliferation index is calculated as the ratio of WST-1 absorbance of treated cells to WST-1 absorbance of control cells.