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.

Cell lines

3T3-L1 fibroblasts, AML-12 mouse hepatocytes

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

0.1, 1, 10, or 50 μM for 1-3 h

Applications

Atglistatin showed highest adipose triglyceride lipase (ATGL) inhibition potential with IC50 value of 0.7 μM. Atglistatin was highly effective in inhibiting lipolysis in 3T3-L1 adipocytes and white adipose tissue (WAT) organ cultures of wild-type mice by targeting ATGL.

Animal models

Mice (C57Bl/6J) model

Dosage form

200 μmol/kg; oral gavage or intraperitoneal administration, for 8 h;

Applications

Atglistatin showed the potential of inhibiting lipolysis in fasted wild-type C57Bl/6J mice. Atglistatin also caused a strong reduction in plasma triacylglycerol (TG) levels (-43%).

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

Atglistatin is a potent and selective inhibitor of ATGL with IC50 value of 0.7 μM [1].

Adipose triglyceride lipase (ATGL) is a rate-limiting enzyme in the mobilization of fatty acids from cellular triglyceride stores. Deregulated fatty acid metabolism may lead to metabolic disorders and dyslipidemic [1].

Atglistatin is a potent and selective ATGL inhibitor. In lysates from E. coli overexpressing ATGL, atglistatin inhibited ATGL activity with IC50 value of 0.7 μM. Atglistatin at concentrations up to 50 μM had no cytotoxicity. Also, atglistatin inhibited ATGL with Ki value of 355 nM in a competitive way. In white adipose tissue (WAT) lysates of wild-type mice, atglistatin inhibited triacylglycerol (TG) hydrolase activity by 78%. The combination of Atglistatin and the HSL inhibitor Hi 76-007917 inhibited TG hydrolase activity by 95%. In WAT cultures of wild-type mice, atglistatin reduced FA and glycerol release up to 72% and 62% respectively in the presence of forskolin [1].

In fasted wild-type C57Bl/6J mice, atglistatin inhibited lipolysis in a dose- and time-dependent way. Atglistatin reduced plasma TG levels by 43% and decreased glycerol and FA levels up to 62% and 50% respectively in a dose dependent way [1].

Reference:
[1].  Mayer N, Schweiger M, Romauch M, et al. Development of small-molecule inhibitors targeting adipose triglyceride lipase. Nat Chem Biol, 2013, 9(12): 785-787.