Application of isopentenyl flavonoid compounds in preparation of antidiabetic drugs
A technology of prenyl flavonoids and compounds, applied in the field of medicine, to achieve the effect of strong inhibitory activity
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Embodiment 1
[0030] Embodiment 1 extracts the compound of the present invention from mulberry
[0031] (1) Extraction: after pulverizing 10 kilograms of stems and branches of the tree, extract by percolation with 200 liters of 95% aqueous ethanol, collect the percolation liquid, and concentrate under reduced pressure to obtain 380 grams of extract. The extract was dissolved in water and extracted with chloroform, and the recovered solvent was concentrated to dryness to obtain 140 g of chloroform extract;
[0032] (2) Separation: 140 grams of chloroform extract was subjected to MCI gel CHP-20P column chromatography, and methanol-water (50:50→60:40→70:30→80:20→90:10→100:0) Gradient elution, each gradient consumption 10 liters, collect fraction; Wherein methanol-water 70: 30 fraction (6.3 grams) and 80: 20 fraction (4.4 grams) carry out Sephadex LH-20 gel column chromatography separation respectively, Eluted with methanol, and combined by thin layer chromatography to obtain 17 fractions: 1-1...
Embodiment 2
[0038] Example 2 Determination of the inhibitory activity of the compound (I) of the present invention on protein tyrosine phosphatase 1B (protein tyrosinephosphatase 1B, PTP1B)
[0039] (1) Preparation of protein tyrosine phosphatase 1B protein
[0040] The recombinant plasmid containing PTP1B catalytic domain was transformed into Escherichia coli BL21-CodonPlus (DE3) for expression. Inoculate the BL21-CodonPlus (DE3) cells containing the recombinant plasmid on the Luria-Bertani (LB) medium containing 100 mg / l ampicillin, culture with shaking at 37 °C, and when the cell concentration reaches 600 nm, the absorbance value is 0.4 Between -0.6, add isopropylthiogalactoside (IPTG) to 500nM to induce the expression of protein; carry out the purification of protein sample with GSTrap FF column, carry out protein analysis with 10% polyacrylamide gel electrophoresis, protein The concentration was determined by the Coomassie brilliant blue method (Bradford), with bovine serum albumin ...
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