Iridoid compound and its preparation method and application
A technology for iridoids and compounds, applied in the field of iridoids and their preparation and application, to achieve good anti-tumor activity, novel structure, and simple extraction and separation methods
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Embodiment 1
[0050] Using 0.8kg of dried baobab fruit as raw material, according to the solid-liquid ratio of 1:8 (kg / L), reflux extraction with 70% ethanol three times, each time for one hour, concentrate to obtain 75g of extract-like ethanol extract, and concentrate to obtain The ethanol extract is suspended in water, and after extraction with n-butanol (3 times, 750ml / time), the extract part extracted by n-butanol is separated by medium and low pressure preparative chromatography (C18 filler, particle diameter 50um, column length 50cm, diameter 8cm), using methanol-water (methanol volume content is 10%, 40%, 70%) sequential gradient elution, flow rate: 30mL / min, the amount of eluent for each gradient is 5 times the column volume, 40% of which is taken The eluted part of methanol is separated by C18 packing open column chromatography (C18 packing, particle diameter 150um, column length 120cm, diameter 10cm), using methanol-water (methanol volume content is 10%, 30%, 60%, 95%) sequentially...
Embodiment 2
[0067] Using 0.8kg of dried baobab fruit as raw material, according to the solid-liquid ratio of 1:8 (kg / L), reflux extraction with 70% ethanol three times, each time for one hour, concentrate to obtain 75g of extract-like ethanol extract, and concentrate to obtain The ethanol extract is suspended in water, and after extraction with n-butanol (3 times, 750ml / time), the extract part extracted by n-butanol is separated by medium and low pressure preparative chromatography (C18 filler, particle diameter 50um, column length 50cm, diameter 8cm), use methanol-water (methanol volume content is 10%, 40%, 70%) sequential gradient elution, flow rate: 40mL / min, the amount of eluent for each gradient is 4 times the column volume, take 10% of it The eluted part of methanol was separated by semi-preparative high-performance liquid chromatography (Agilent XDB-C18 reversed-phase column-5μm, 250×10mm), and eluted with acetonitrile-water (volume ratio: 10:90) to obtain the compound of formula 2 ...
Embodiment 3
[0084] In vitro growth inhibitory experiments of compounds of formula 1, 2, 3, and 4 on human breast cancer MDA-MB-231 cells: MDA-MB-231 cell monolayers were inoculated in a mixture containing 2% glutamine and 1.5% sodium bicarbonate , 10% fetal bovine serum in RPMI-1640 culture medium. And added 100 units / ml of penicillin and 100 μg / ml of streptomycin. At a temperature of 37°C, CO 2 Cultured in a cell culture incubator at a concentration of 5%. Cells in the logarithmic growth phase were prepared as 1 × 10 4 Inoculate the concentration of cell / mL on a 96-well plate, 0.1mL per well, and then add medium containing different concentrations into the wells, each concentration has 3 parallel groups, add the same amount of solvent to the control group, and place in a 37°C carbon dioxide incubator Culture in medium for 72h, then centrifuge (1000rpm, 20min), discard the supernatant, add 0.20mg / mL MTT serum-free medium to each well, continue to culture at 37°C for 3h, centrifuge, rem...
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