Atglistatin is a highly effective, specific and competitive ATGL inhibitor with an IC50 of ~0.7 μM for inhibition of lipolysis in vitro, and no toxicity when the concentration up to 50 μM.

ATGL:0.7 μM

Atglistatin inhibits ATGL, consequently suppressing lipid breakdown in both cellular and organ cultures without exhibiting cytotoxic effects at concentrations up to 50 µM.

Atglistatin, administered orally (p.o.), reduces fatty acids (FA) and glycerol by up to 50% and 62% respectively, in a dose-dependent manner, and also significantly reduces triglyceride (TG) levels in the plasma by 43%. When administered via intraperitoneal injection (i.p.), Atglistatin inhibits lipolysis in a dose- and time-dependent manner. Moreover, Atglistatin exhibits significant differences in tissue distribution, primarily accumulating in the liver and adipose tissue.

Determination of lipase activity: For determination of lipase activity, lysates are incubated with a substrate containing radiolabeled [9,10-3H(N)]-triolein. Subsequently, FA are extracted and quantitated by liquid scintilation counting. Data are presented as mean S.D. of triplicate determinations and representative for at least three independent experiments.

For MTT-based in vitro viability assays, cells are seeded in 96-well plates and cultured under standard conditions for 24 h. The next day, cells are pretreated with different concentrations of Atglistatin dissolved in DMSO or Cisplatin dissolved in DMF as positive control for 2 h. Medium is replaced by an identical fresh medium and incubated again for the indicated time points. Thereafter, cells are incubated for 3 h with 100 μL Thiazolyl Blue Tetrazolium Bromide (MTT). The resulting violet formazan crystals are dissolved by adding 100 μL of MTT solubilization solution (0.1% NP-40, 4 mM HCl and anhydrous isopropanol). After complete dissolution of the formazan product, absorbance is measured at 595 nm using 690 nm as reference wavelength.(Only for Reference)