HJ-4 is a derivative of Piperine. It effectively inhibits the proliferation of colorectal cancer (CRC) cells by reducing colony formation and DNA synthesis in a dose-dependent manner. HJ-4 notably impairs the adhesion, migration, and invasion of CRC cells and induces apoptosis. In a chicken embryo chorioallantoic membrane (CAM) model with implanted HCT116/SW480 tumor spheroids, HJ-4 demonstrates antitumor efficacy. HJ-4 is applicable for colorectal cancer (CRC) research.

HJ-4 inhibits the viability of colorectal cancer (CRC) cells, with IC50 values of 15.82 μM for HCT116 cells and 20.46 μM for SW480 cells, when applied at concentrations of 3.125-100 μM over 24 hours. At a range of 8-32 μM for 48 hours, HJ-4 reduces both the number and size of colonies in HCT116 and SW480 cells. Additionally, in EdU incorporation assays, HJ-4 at 8-32 μM over 24 hours decreases the number of EdU-positive HCT116/SW480 cells, thereby inhibiting DNA synthesis in a dose-dependent manner. HJ-4, at concentrations of 8-32 μM for 0-48 hours, suppresses adhesion, migration, and invasion of these cells. It also upregulates the expression of p53, PUMA, BAX, cleaved caspase-3, cleaved PARP, and downregulates BCL-2, β-catenin, Cyclin D1 in HCT116/SW480 cells when administered at 8-32 μM for 24 hours. Furthermore, HJ-4 increases p53/BAX expression and promotes early (YO-PRO-1 positive) and late (PI positive) apoptosis in HCT116 cells through YO-PRO-1/PI staining when applied at 8-32 μM for 24 hours.

HJ-4 (8-32 μM, applied locally to the tumor site once daily for 7 days) inhibits angiogenesis in the chicken embryo chorioallantoic membrane (CAM) model implanted with HCT116/SW480 tumor spheroids.