Compounds for modulating FXR activity and uses thereof
A compound and active technology, applied in the field of compounds that regulate FXR activity and their preparation
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Embodiment 1
[0067] Preparation of 6-((3-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)quinolin-7-yl)oxy)fume acid (compound 1)
[0068]
[0069] (a) Compound 1A-2 (1.0g, 4.46mmol, 1.0eq), compound 1A-1 (1.55g, 4.46mmol, 1.0eq) and cesium carbonate (2.90g, 8.92mmol, 2.0eq) were dissolved in DMF ( 10ml), stirred at 65°C for 2 hours. After cooling to room temperature, 10 ml of water and 10 ml of EA (ethyl acetate) were added, and the organic phase was concentrated to dryness to obtain compound 1A-3 (1.61 g, yield: 73.5%, which was used in the next step without further purification) , namely: 4-(((6-bromoquinolin-2-yl)oxy)methyl)-5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazole, LCMS (ESI ): Calculated as C 22 h 15 BrCl 2 N 2 o 2 ;[M+H] + : 489.0, measured value: 489.0.
[0070]
[0071] (b) Compound 1A-3 (200mg, 0.408mmol, 1.0eq) was dissolved in anhydrous THF (2ml), and then 2 Add KOAc (80mg, 0.816mmol, 2.0eq), Pd(dppf) 2 Cl 2 (30 mg, 0.408 mmol, 0.1 equiv) and ...
Embodiment 2
[0080] Preparation of 6-(((7-((5-cyclopropyl-3-(2,6-dichlorophenyl)isoxazol-4-yl)methoxy)quinolin-3-yl)oxy) Niacin (Compound 2)
[0081]
[0082] (a) Compound 2B-1 (1.0g, 4.46mmol, 1.0eq), compound 1A-6 (0.96g, 4.46mmol, 1.0eq) and cesium carbonate (2.90g, 8.92mmol, 2.0eq) were dissolved in DMF ( 10ml), stirred at 65°C for 2 hours. After being cooled to room temperature, add 10ml water and 10ml EA (ethyl acetate), the organic phase is concentrated to dryness, obtains compound 2B-2 (1.1g, productive rate: 68.7%), namely: 6-((7-bromoquinone Methyl nicotinate, LCMS (ESI): calculated as C16H11BrN2O3; [M+H] + : 359.0, measured value: 359.0.
[0083]
[0084] (b) Compound 2B-2 (200mg, 0.56mmol, 1.0eq) was dissolved in anhydrous THF (2ml), and then heated under N 2 Add KOAc (110 mg, 1.12 mmol, 2.0 equiv), Pd(dppf) 2 Cl 2 (41mg, 0.056mmol, 0.1eq) and bis(pinacolate)diboron (142mg, 0.56mmol, 1.0eq), the reaction mixture was heated to reflux for 2h. After cooling to room te...
Embodiment A
[0094] Example A: FXR Agonist Binding Capability
[0095] In this example, the analysis of the binding activity of compounds 1 and 2 prepared in Examples 1 and 2 to FXR was carried out using a time-resolved fluorescence resonance energy transfer coactivator peptide recruitment experiment. The assay uses LanthaScreen-labeled anti-GST antibody, fluorescein-labeled SRC2-2 coactivator peptide, and GST-tagged FXR-LBD. LanthaScreen-labeled anti-GST antibody indirectly labels FXR-LBD by binding to the GST tag. FXR-LBD can form heterodimer FXR-LBD / RXRα with RXRα, and the compound can bind to FXR-LBD / RXRα and cause a conformational change of FXR-LBD, thereby increasing the effect of heterodimer on SRC2-2 coactivation Peptide recruitment capacity. At the same time, this results in a tighter binding of FXR-LBD (labeled with GST) and SRC2-2 coactivator peptide, and then adding anti-GST antibody (labeled with LanthaScreen), LanthaScreen reacts with fluorescein to produce fluorescence. ...
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