Steroid compound agonist for PPARgamma and its purpose
A use and composition technology, applied in the direction of medical preparations containing active ingredients, drug combinations, metabolic diseases, etc., can solve problems such as increased heart attack rate, weight gain, side effects and limited development
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0061] Example 1, demonstrates that RU-486 is a potent PPARγ ligand.
[0062] To demonstrate that the steroid ligand RU-486 can activate nuclear receptors, we co-transfected Cos7 cells with a Gal-4-driven reporter gene and a plasmid containing the gene of interest. In comparison with previous observations, RU-486 exhibited low glucocorticoid receptor (GR) activating activity in the absence of endogenous and exogenous glucocorticoids. It was evident that the transcriptional activity of PPARγ was significantly induced in RU-486-treated cells (Fig. 1A). RU-486 induced PPARγ activity was verified by full-length sequence protein and upstream protein experiments (Fig. 1B). The curve results of the above experiments show that the activation effect of RU-486 on PPARγ is related to the concentration of its own drug, and it is found that the EC50 (half effective dose) of RU-486 is similar to that of rosiglitazone, which means that RU-486 is a A potent PPARγ ligand.
[0063] The prese...
Embodiment 2
[0064] Example 2, Molecular Structure Verification Experiment of the Complex of PPARγ Ligand Binding Region Fragment and RU-486
[0065] In order to detect the high binding affinity between RU-486 and PPARγ at the molecular level, we solved the resolution up to 2.5 Protein crystal structure of the PPARγ / RU-486 complex with the coactivator SRC1-2LXXLL motif. The structure shows that RU-486 binds to the PPARγ ligand binding domain in a classical conformation, as rosiglitazone has a similar structure to PPARγ (Fig. 3A).
[0066] The structures of PPARγ / RU-486 and PPARγ / rosiglitazone showed similarities, and their ligands stayed at the same binding site in the ligand pocket of PPARγ (Fig. 6A & 6B). Overlapping the two structures shows that the steroid core of RU-486 overlaps with the pattern of rosiglitazone, which indicates that the steroid core fully occupies the ligand binding site of PPARγ (Fig. 3B). A distinct dimethylaniline side chain of RU-486 was seen from the high res...
Embodiment 3
[0067] Example 3, Unique Binding Model of RU-486 in the PPARγ Ligand Pocket
[0068] In order to verify the role of ligand pocket residues in binding to RU-486 and the activity of PPARγ, we mutated several key residues in contact with RU-486, and then detected the transcriptional activity of these mutated PPARγ. R288, A292, and I236 are three important residues on the ligand pocket that bind to the dimethylaniline side chain of RU-486 (Fig. 4A). The results indicated that the interaction between the dimethylaniline side chain of RU-486 and the ligand pocket of PPARγ is the prerequisite for the stable binding of RU-486 to PPARγ.
[0069] The 17β-hydroxyl group of the steroid core forms hydrogen bonds with some amino acid residues surrounding PPARγ, including residue Y327 in helix 5 and residue H449 in helix 10 (Fig. 3D). Both RU-486 and rosiglitazone bind to these binding sites on the PPARγ ligand pocket, thereby supporting a key conserved mechanism required for ligand-mediate...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More