In vitro activity: Thiomyristoyl, a thiomyristoyl lysine compound, is a novel, potent and specific SIRT2 inhibitor with IC50 of 28 nM. Thiomyristoyl showed broad-spectrum of anticancer activity in various human cancer cells and mouse models of breast cancer but has little effect on non-cancerous cells. Targeting sirtuins for cancer treatment has been a topic of debate due to conflicting reports and lack of potent and specific inhibitors. The anticancer effect of Thiomyristoyl correlates with its ability to decrease c-Myc level. Thiomyristoyl had limited effects on non-cancerous cells and tumor-free mice, suggesting that cancer cells have an increased dependency on SIRT2 that can be exploited for therapeutic benefit. Thiomyristoyl is a promising anticancer agent and may represent a general strategy to target certain c-Myc-driven cancers. Thiomyristoyl(TM) is a highly selective SIRT2 inhibitor. It cannot efficiently inhibit SIRT3, SIRT5, SIRT6, or SIRT7. In vitro, it shows great inhibition of cell viability and its cytotoxicity is relatively selective toward cancer cells. TM decreases c-Myc oncoprotein level in cancer cells, the ability of TM to decrease c-Myc abundance in different cell lines correlates with the sensitivity of the cell lines to TM

Kinase Assay: Thiomyristoyl, a thiomyristoyl lysine compound, is a novel, potent and specific SIRT2 inhibitor with IC50 of 28 nM. It cannot efficiently inhibit SIRT3, SIRT5, SIRT6, or SIRT7.

Cell Assay: Cells are seeded into 96-well plates at 3,000–4,000 cells per well. After 24 hr, test compounds (Thiomyristoyl) are added to cells to final concentrations ranging from 1 to 50 μM. Cells are then incubated for 72 hr and cell viability is measured using the CellTiter-Blue viability assay. Relative cell viability in the presence of test compounds is normalized to the vehicle-treated controls after background subtraction. GraphPad Prism software is used to determine the IC50 values. Knockdown of SIRT1-7 in various cell lines is achieved by lentiviral infection.