Cyclovirobuxine D (Bebuxine) is extracted from Buxus microphylla.

The LD50 of Cyclovirobuxine D in mice is found to be 8.9 mg/kg (i.v.), 9.2 mg/kg (i.p.), and 293 mg/kg (p.o.). This compound is observed to reduce the weight of venous thrombosis in rats. In anesthetized pigs, Cyclovirobuxine D initiates coronary vasodilation, a mechanism associated with the release of nitric oxide in endothelial cells. Furthermore, it also improves cardiac failure induced by myocardial infarction in rats.

Cyclovirobuxine D facilitates the utilization of intracellular Ca(2+) and prevents its efflux, thereby exerting protective effects against heart failure. It enhances the vitality of cardiomyocytes damaged by oxidation or hypoxia. Furthermore, it significantly reduces the infarct size caused by ligation of the coronary artery in rats. Cyclovirobuxine D also protects the endothelial cells of rat aortas from hypoxic damage and increases the release of NO within these cells.

MDA-MB-231 cells treated as indicated or tumor tissues are harvested and lysed in Mg2+ lysis buffer containing 50 mM Tris (pH 7.5), 10 mM MgCl2, 0.5 M NaCl, and protease inhibitor cocktail. Equal amounts of lysates are incubated with PAK-PBD beads at 4°C for 1 h. PAK-PBD beads are pelleted by centrifugation and washed with ish buffer containing 25 mM Tris (pH 7.5), 30 mM MgCl2, 40 mM NaCl. Active Rac1 is detected by western blotting.