Bioymifi (DR5 Activator) is an effective activator of the TRAIL receptor DR5, binding to the extracellular domain (ECD) of DR5 with a Kd of 1.2 μM. Bioymifi can serve as a novel mimetic of TNF-related apoptosis-induced ligand (TRAIL), inducing DR5 aggregation as a sole inducer, and stimulating cell apoptosis.

DR5 receptor:1.2 μM(Kd)

方法：红细胞用 Bioymifi (10-100 µM) 处理 24 h，通过 flow cytometry 鉴定 Bioymifi 的溶血和杀菌特性及潜在分子机制。
结果：Bioymifi 能发挥剂量反应性、钙非依赖性溶血作用，减少红细胞血红蛋白，显著增加膜联蛋白 V、Fluo4 和 DCF 阳性细胞，以及对前向和侧向光散射的双重影响。[1]
方法：新生大鼠心室肌细胞 (nrvm) 用 Bioymifi (10 µM) 处理 24 h，通过 Western Blot 检测靶点蛋白表达水平。
结果：Bioymifi 处理的 nrvm 中 DR5 的表达高于 NRVF。[2]

方法：为研究 DR5 激活在心脏中的作用，将 Bioymifi (5 mg/kg/d) 微型渗透泵给药给 C57BL/6 小鼠，每天一次，持续两周。
结果：Bioymifi 的给药导致心脏器官和心肌细胞水平的肥大，但并不影响心肌细胞的死亡或心脏纤维化。[2]

Solubilized membranes from 3T3 Flk-1 cells are added to polystyrene ELISA plates that has been precoated with a monoclonal antibody that recognizes Flk-1. After an overnight incubation with lysate at 4°C, serial dilutions of SU5416 are added to the immunolocalized receptor. To induce autophosphorylation of the receptor, various concentrations of ATP are added to the ELISA plate wells containing serially diluted solutions of SU5416. The autophosphorylation is allowed to proceed for 60 min at room temperature and then stopped with EDTA. The amount of phosphotyrosine present on the Flk-1 receptors in the individual wells is determined by incubating the immunolocalized receptor with a biotinylated monoclonal antibody directed against phosphotyrosine. After removal of the unbound anti-phosphotyrosine antibody, avidin-conjugated horseradish peroxidase H is added to the wells. A stabilized form of 3,3′,5,5′-tetramethyl benzidine dihydrochloride and Water2 is added to the wells. The color readout of the assay is allowed to develop for 30 min, and the reaction is stopped with H2SO4. Parallel biochemical kinase assays are performed to measure autophosphorylation on EGFR and fibroblast growth factor receptor[1].

Dose-response curves of bioymifi and A2C2 in T98 g cells are plotted as a function of A2C2 or bioymifi concentration. Human glioblastoma (T98 g) cells are treated with various concentrations of A2C2 or bioymifi alone or in combination with 1 μM Smac mimetic (SM) for 48 h. The corresponding cell survival is normalized to the treatment without A2C2 or bioymifi.(Only for Reference)