In vitro activity: Succinobucol (formerly known as AGI-1067, AGZ-1067, Probucol monosuccinate, succinyl ester of probucol) is a a lipid-lowering compound and a phenolic antioxidant with anti-inflammatory and antiplatelet effects. Succinobucol inhibits TLR4 ligand (LPS)-induced activation of ASK1 and the downstream p38 and JNK MAP kinases, which could inhibit ASK1-JNK/p38-dependent gene expression of proinflamamtory molecules. Inhibition of both p38 and JNK by AGI-1067 occurred in a concentration-dependent manner with an IC50 value under 5 μM. AtheroGenics reports positive results from ANDES Phase 3 Clinical Trial (NCT00066898) of Succinobucol.

Kinase Assay: Succinobucol (10, 50, 100 μM) causes a dose-dependent reduction in collagen-induced platelet aggregation in rabbit whole blood. Succinobucol also causes a significant reduction in whole blood aggregation in response to ADP. Succinobucol (10, 100 μM) significantly lowers the relaxation to X/XO. Succinobucol significantly prevents 3-NP-induced loss of SH-SY5Y cell viability, generation of reactive oxygen species, and decrease of ΔΨm. Succinobucol does not protect against 3-NP-induced inhibition of mitochondrial complex II activity, pointing to the mitigation of secondary events resultant from mitochondrial complex II inhibition. Succinobucol significantly increases (50 %) the levels of GSH in SH-SY5Y cells, which is paralleled by significant increases in glutamate cysteine ligase messenger RNA (mRNA) expression and activity. Succinobucol effectively exhibits superior inhibitory effects on cell migration and invasion activities, VCAM-1 expression and cell-cell binding of RAW 264.7 to 4T1 cells. Succinobucol also shows inhibitory effect on VCAM-1 expression in 4T1 cells and cell-cell binding of RAW 264.7 to 4T1 cancer cells.

Cell Assay: The cytotoxicity of Succinobucol is determined in the metastatic 4T1 breast cancer cells. Cells are added to 96-well plates at 6×103 cells/well and cultured overnight. Then, Succinobucol, SCB and the PCD polymer (equivalent concentration to SCB) are respectively added to each well with SCB concentrations ranging from 4 ng/mL to 40 μg/mL. Cells without any treatment are performed as negative control. Thereafter, cells are incubated for further 48 h, and the cell viability is measured by MTT assay method.