GW9662 (TIMTEC-BB SBB006523) is a specific PPARγ antagonist (IC50: 3.3 nM, in a cell-free assay), with 100 to 1000-fold functional selectivity for PPARγ than PPARα/δ in cells.

PPARγ:3.3 nM

GW9662抑制PPARγ激活,并抑制人乳腺癌肿瘤细胞系（MCF7, MDA-MB-468, MDA-MB-231）生长（IC50：20 -30 μM）。GW9662结合PPARγ上的Cys(285),其是三种PPAR中较保守的。在MDA-MB-231细胞中,Rosiglitazone（50 μM）与GW9662（10 μM）联用7天后,其存活细胞数的减少程度具有统计学意义。在原代鼠骨髓和RAW264.7细胞中,PPARγ1配体对RANKL诱导的破骨细胞形成有抑制作用,而GW 9662（2 μM）可浓度依赖性地逆转这些配体的抑制作用。在RAW264.7细胞中,GW 9662（1 μM）抑制NF-κB的RANKL激活。 在BMs中,GW 9662（2 μM）阻断IL-4对破骨细胞形成的抑制。在甲状腺眼病患者的原代前脂肪细胞中,GW9662（10 μM）对激素和激动剂诱导的脂肪细胞分化有抑制作用。

大鼠经脂多糖（1 mg/kg,i.p.）预处理,可明显减弱肾损伤和功能障碍引起的所有缺血/再灌注损伤特征.而GW9662（1 mg/kg,i.p.）可阻断脂多糖的保护作用.

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)