Novel compounds and their use in treatment of autoimmune diseases
A compound and free technology, applied in the direction of allergic diseases, metabolic diseases, skin diseases, etc., can solve the problems of destroying normal tissues, and the immune system cannot distinguish harmful antigens in body organs, so as to restore immune balance and damaged tissues, prevent and treat diseases The effect of treating autoimmune diseases
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[0183] 1. Synthesis of N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide (compound 8)
[0184] [Process 1]
[0185]
[0186] Stir 2-(5-chloro-1H-indol-3-yl)acetic acid (1.00 g, 4.77 mmol) in CH at room temperature 2 Cl 2 (30 mL), 5-bromo-6-methylpyridin-2-amine (892 mg, 4.77 mmol), 1-[bis(dimethylamino)methylene]-1H-1 were added dropwise in this order. , 2,3-triazo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 2.18 g, 5.72 mmol) and trimethylamine (1.33 mL, 9.54 mmol). The reaction mixture was stirred at room temperature for 3 days, and distilled water (10 mL) was added to the mixture to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4 Dry and filter. After the filtrate was concentrated under reduced pressure, the concentrate was subjected to column chromatography (SiO 2 , Hexane:EtOAc = 4:1-2:1) was purified to give the light grey compound (970 mg, 54%)....
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[0338] Example: Measurement of the activity of compounds - experimental protocol
[0339] 1. Compound Search and Preparation
[0340] In order to confirm the target specificity of the prepared compound, evaluation was performed by the following method.
[0341] After recovering HepG2 cultured in DMEM-fetal bovine serum (FBS) 10% medium and confirming the viability of 97% or more by trypan blue staining, the recovered product was centrifuged at 1200 rpm for 5 minutes at room temperature, and by dividing the cells by 3 × 10 5 Cells / ml were prepared by resuspending in DMEM-fetal bovine serum 10% medium. Thereafter, cells were aliquoted at 3 mL into 60 mm dishes, and each dish was treated with 50 μl of compound at a concentration of 5 μM diluted in DMEM medium, and then incubated in a cell incubator (5% CO 2 incubator) for 24 hours. As a control, 50 μl of 0.05% dimethyl sulfoxide (DMSO) / DMEM medium was used for treatment.
[0342] The cultured cells were recovered to prepar...
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