In vitro activity: T0901317 acts through LXR and in concert with its RXR heterodimerization partner induces the expression of the ABCA1 reverse cholesterol transporter. T0901317 upregulates expression of the ABCA1 gene associated with cholesterol efflux regulation and HDL metabolism. T0901317 displays an EC50 of ~ 5 μM for activation of bile acid farnesoid X receptors (FXRs), 10-fold more potent than natural FXR ligand chenodeoxycholic acid. T0901317, is also a high-affinity ligand for the xenobiotic receptor pregnane X receptor (PXR). T0901317 binds and activates PXR with the same nanomolar potency with which it stimulates LXR activity. T0901317 induces expression not only of LXR target genes, but also of PXR target genes in cells and animals, including the scavenger receptor CD36. T0901317 decreases amyloid-β production in primary neurons in vitro. T0901317 is found to directly bind to RORα and RORγ with high affinity (Ki = 132 and 51 nM, respectively), resulting in the modulation of the receptor's ability to interact with transcriptional cofactor proteins. T0901317 represses RORα/γ-dependent transactivation of ROR-responsive reporter genes and in HepG2 cells reduces recruitment of steroid receptor coactivator-2 by RORα at an endogenous ROR target gene (G6Pase).

Kinase Assay: The LXR ligand-binding domain was fused to the C-terminus of glutathione S-transferase (GST) and the resultant GST-LXR protein was expressed in Escherichia coli and purified on glutathione beads. Rhodamine-labeled peptide (10 nM; with amino acid sequence ILRKLLQE) was incubated on a shaker for 1 h with 400 nM GST-LXR and T0901317 in 100 μL of buffer (10 mM Hepes, 150 mM NaCl, 2 mM MgCl2, 5 mM DTT at pH 7.9) in a 96-well plate. Fluorescence polarization (mP) was measured on an LJL analyst (LJL Biosystems).

Cell Assay: T0901317 is a highly potent and selective nonsteroidal LXR ligand. T0901317 induces transcriptional activity of LXRα nearly 8-fold with EC50 value of 20 nM. T0901317 also transactivates chimeric Gal4-LXRα and Gal4-PXR (pregnane X receptor).

Science. 2000 Sep 1;289(5484):1524-9; J Biol Chem. 2005 Feb 11;280(6):4079-88.