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)

METHODS: Erythrocytes were treated with Bioymifi (10-100 µM) for 24 h. The hemolytic and bactericidal properties and potential molecular mechanisms of Bioymifi were characterized by flow cytometry.
RESULTS: Bioymifi exerted a dose-responsive, calcium-independent hemolytic effect, reduced erythrocyte hemoglobin, significantly increased membrane-bound protein V, Fluo4, and DCF-positive cells, as well as a dual effect on forward and lateral light scatter. [1]
METHODS: Neonatal rat ventricular myocytes (nrvm) were treated with Bioymifi (10 µM) for 24 h, and the expression levels of target proteins were detected by Western Blot.
RESULTS: The expression of DR5 in Bioymifi-treated nrvm was higher than that of NRVF.[2]

METHODS: To investigate the role of DR5 activation in the heart, Bioymifi (5 mg/kg/d) micro-osmotic pump was administered to C57BL/6 mice once daily for two weeks.
RESULTS: Administration of Bioymifi resulted in hypertrophy at the level of cardiac organs and cardiomyocytes, but did not affect cardiomyocyte death or cardiac fibrosis. [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)