Application of diterpenoid compound
A compound and diterpenoid technology, applied in the application field of diterpenoids, can solve the problems of aggravation of the patient's condition, functional decline, drug failure, etc., and achieve a significant anti-islet cell apoptosis effect.
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Embodiment 1
[0027] The preparation of embodiment 1 compound of the present invention
[0028] Take 10 kg of wild licorice, extract with ethanol reflux, dissolve the extract in water after concentrating the extract, then extract with petroleum ether, recover the upper organic phase, and extract the remaining water layer with dichloromethane, recover the lower organic phase, and concentrate under reduced pressure , to obtain dichloromethane extract (180g).
[0029] Get the obtained dichloromethane extract, use silica gel dry method to mix the sample, add organic solvent to dissolve the dichloromethane extract, mix well with silica gel particles, after the organic solvent evaporates to dryness, grind finely, use silica gel column chromatography (silica gel 200 -300 mesh, particle size is 200-300μM), gradient elution is carried out with a mixed solution of petroleum ether and acetone, and the fraction obtained by using a volume ratio of petroleum ether and acetone of 100:16 is applied to a si...
Embodiment 2
[0035] Embodiment 2: Anti-apoptotic activity experiment of compounds of the present invention in MIN-6 cells
[0036] Methods: (1) Resurrection and culture of MIN6 cells: Take them out from the refrigerator at -80°C, centrifuge to discard the upper cryopreservation solution, add 1640 medium containing 10% fetal bovine serum and 1% double antibody to resuspend the cells, and the relative humidity is Culture in an incubator at 95%, 37°C, 5% CO2; depending on the growth of the cells, replace the fresh medium every 24-36 hours, and pass passage or freeze when the cells grow to 75%-85%. (2) Diterpenoid intervention model group (PA) culture: MIN6 cell proliferation grouping: well-grown MIN6 cells were evenly inoculated into 96-well plates, and divided into 5×10 5 100 μLC complete medium was added per mL. The cells were divided into the following groups: blank control group (CK), 300 μg / mL LC model group (PA), compound Ⅰ 25 μg / mL dosing group, 50 μg / mL dosing group. The original cu...
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