GW9662 (TIMTEC-BB SBB006523) is a PPARγ antagonist (IC50=3.3 nM) with selectivity. GW9662 can be used to study the pathogenesis of metabolic diseases, such as obesity and diabetes, by inhibiting the activity of PPARγ. GW9662 can be used to study the pathogenesis of inflammatory diseases, such as atherosclerosis and rheumatoid arthritis. GW9662 has anti-tumor effect.

PPARα:32 nM, MDA-MB-231 cells:20-30 µM, MCF7 cells:20-30 µM, PPARδ:2000 nM, PPARγ:3.3 nM, MDA-MB-468 cells:20-30 µM

METHODS: Human breast cancer cell lines (MCF7, MDA-MB-468, and MDA-MB-231) were treated with GW9662 (100 nM-50 mM) for 72 hours, and MTT assay was used to detect the inhibition of cell growth.
RESULTS: GW9662 significantly inhibited the proliferation of MCF7, MDA-MB-468, and MDA-MB-231 cells (IC50= 20-30 μM). [1]

METHODS: To study the blocking effect of GW9662 on the protective effect of lipopolysaccharide, first, rats were pretreated with lipopolysaccharide (1 mg/kg,i.p.), which could significantly weaken all ischemia/reperfusion injury characteristics caused by renal injury and dysfunction. Then, GW9662 (1 mg/kg) was intraperitoneally injected into the rats.
RESULTS: GW9662 can block the protective effect of lipopolysaccharide.[2]

Binding assay: The human PPARα, PPARγ, and PPARδ ligand binding domains (LBDs) are expressed in E. coli as polyhistidine-tagged fusion proteins. Receptors are immobilized on SPA beads by addition of the desired receptor (15 nM) to a slurry of streptavidin-modifed SPA beads (0.5 mg/mL) in assay buffer. The mixture is allowed to equilibrate for at least 1 hour at room temperature, and the beads are pelleted by centrifugation at 1×103 g. The supernate is discarded, and the beads are resuspended in the original volume of fresh assay buffer with gentle mixing. The centrifugation/resuspension procedure is repeated, and the resulting slurry of receptor-coated beads is used immediately or stored at 4 ℃ for up to 1 week before use. [3H]GW2443 are used as radioligands for determination of competition binding to PPARα, PPARγ, and PPARδ, respectively. Unless otherwise indicated, the buffer used for all assays is 50 mM HEPES (pH 7), 50 mM NaCl, 5 mM CHAPS, 0.1 mg/mL BSA, and 10 mM DTT. For some experiments, the HEPES (pH 7) is replaced with 50 mM Tris (pH 8).

MDA-MB-231 cells are seeded at a density of 1 × 105 cells per 25 cm3 tissue culture flask. After 24 h (day 0), the growth medium is replaced with fresh medium containing rosiglitazone (50 μM), GW9662 (10 μM) or both together. Control flasks receives 0.1% DMSO. Cells are harvested on days 0, 3, 5, 7, 10 for each treatment condition by trypsinisation, stained using trypan blue, and the total and viable number of cells per flask calculates using a haemocytometer.(Only for Reference)