Cell experiment:

HepG2 cells are seeded at 5×105 cells/mL in a 4-chamber culture slide. The following day the media is removed and replaced with antibiotic-free media containing 10 μM of DMSO or SR9238 and cells are allowed to grow for 48 h. Cells are washed in TBS, fixed and fluorescently stained for SREBP2 according to the assay protocol[1].

Animal experiment:

Twenty-one-week-old male C57BL6 DIO mice are used. Animals are individually housed and fed a high fat diet (60% kcal/fat diet, 20% carbohydrate) for the duration of the experiment that includes SR9238 administration for 30 days (30 mg/kg, qd, ip). Prior to initiation of the experiment, animals are provided the high fat diet for 10-weeks. Animals are acclimated to the environment for one week and sham dosed with vehicle for 3 days prior to SR9238 administration. Body weight and food intake are monitored daily. Pre- and post-experiment body composition analysis is performed on all the mice. Blood is collected by cardiac puncture and used for plasma cholesterol and triglyceride measurements. Livers are weighed and immediately flash-frozen in liquid nitrogen for gene expression analysis or put in formalin on ice for histology[1].

SR9238 is a selective inverse agonist of LXRα and LXRβ with IC50 values of 214 and 43 nM, respectively [1].

The liver X receptors (LXRα and LXRβ) are members of the nuclear receptor superfamily that act as ligand-dependent transcription factors. LXRα is mainly expressed in the liver, kidneys, intestines, and adipose tissues while LXRβ is ubiquitously expressed. LXRs are well-characterized regulators of the expression of an array of lipogenic enzyme genes [1][2].

SR9238 is a potent and selective inverse agonist of LXRα and LXRβ. SR9238 increased interaction of corepressor NCoR ID1 with LXRα and LXRβ with EC50 values of 33 nM and 13 nM, respectively, and NCoR ID2 with LXRα and LXRβ with EC50 values of >10 μM and 93 nM. SR9238 also effectively inhibited transcription from a Fasn promoter driven luciferase reporter [1].

In mice, SR9238 was detected in the liver 2h after the injection and was not detectable in the plasma, skeletal muscle or brain. SR9238 displayed liver specific exposure. In hepatic steatosis mice, SR9238 (30 mg/kg/day, i.p.) significantly inhibited lipogenic gene expression and reduced lipid content in the liver [1]. In non-alcoholic steatohepatitis mice, SR9238 significantly reduced the severity of hepatic steatosis, reduced hepatic inflammation and ameliorated hepatic fibrosis [2].

References:
[1].  Griffett K, Solt LA, El-Gendy Bel-D, et al. A liver-selective LXR inverse agonist that suppresses hepatic steatosis. ACS Chem Biol. 2013 Mar 15;8(3):559-67.
[2].  Griffett K, Welch RD, Flaveny CA, et al. The LXR inverse agonist SR9238 suppresses fibrosis in a model of non-alcoholic steatohepatitis. Mol Metab. 2015 Feb 9;4(4):353-7.