Extractive of compound formula of ophiopogon decoction and caper euphorbia seed and reed stem soup and application thereof in preparing medicament for inhibiting H460 cell proliferation
A technology of Ophiopogon japonicus soup and extracts, applied in the direction of drug combinations, medical preparations containing active ingredients, antineoplastic drugs, etc.
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Embodiment 1
[0049] Example 1: Materials and methods.
[0050] 1. Main reagents
[0051] Rotary evaporator (Nanjing Jinzheng Teaching Instrument Co., Ltd., 1002 lifting constant temperature water bath), water circulation vacuum pump (Yingyu Yuhua Instrument Factory, Gongyi City, Henan Province), digital display thermostat heating mantle (Zhangzhishan Town, Tongzhou City), HH-S Digital display constant temperature water bath (Jintan Medical Instrument Factory), round bottom flask (20L), condenser tube, iron stand, separatory funnel (5000mL) (Nanjing Jinzheng Teaching Instrument Co., Ltd.).
[0052] Ophiopogon japonicus (Sichuan), French pinellia (Guizhou), sun-dried ginseng (Jilin), licorice (Inner Mongolia), reed stem (Anhui), coix seed (Guizhou), winter melon seeds (Anhui), peach kernel (Shandong).
[0053] Medicinal ethanol (analytical grade, Nanjing Chemical Reagent Co., Ltd.), cyclohexane (analytical grade, Shanghai Shisi Hewei Chemical Co., Ltd.), ethyl acetate (analytical grade, Nan...
Embodiment 2
[0057] Example 2: Preparation of the extract of Maimendong decoction combined with Qianjinweijing decoction.
[0058]Weigh 200g of Ophiopogon japonicus, 100g of Pinellia chinensis, 200g of raw sun-dried ginseng, 80g of licorice, 200g of reed stem, 200g of coix seed, 200g of winter melon seed, 100g of peach kernel, and make up a total of 1280g of the whole formula, crush it and put it in a 20,000mL round bottom flask , add 10 times the amount of medicinal materials in pure water, shake and mix, soak for 30 minutes; connect the reflux condensing device, put it in the electric heating mantle and heat slowly to 110°C to boil, and keep boiling slightly for 2 hours, stop heating, and place it to the solution temperature Cool down to room temperature, remove the reflux condenser, pour out the solution in the flask, and filter it with gauze. Add purified water to the dregs to the same scale as the first time, extract once more in the same way, combine the extracts, and concentrate und...
Embodiment 3
[0059] Example 3: Experiment of the inhibitory effect of the ethyl acetate extraction fraction (MW-06 fraction) on H460 cells.
[0060] 1. Experimental method
[0061] 1.1 Cell proliferation inhibition test (MTT) method
[0062] Take the NCI-H460 cells and HFL-1 cells in the logarithmic growth phase, and adjust the cell concentration to 1×10 4 / mL, inoculated in a 96-well culture plate, 200 μL per well, cultured for 24 hours, aspirated the supernatant, and replaced each well with 200 μL of serum-free medium. After 12 hours, aspirated the culture solution. Add 200 uL of 0.1 μg / mL, 0.5 μg / mL, 1 μg / mL, 5 μg / mL, 10 μg / mL, 50 μg / mL, 100 μg / mL drug-containing culture solution into each well, and add an equal volume of DMSO (final concentration 1‰) culture medium, the positive control group was added with a final concentration of 1 μg / mL equal volume of cisplatin injection (DDP). Set 6 parallel wells in each group. After 72 hours of treatment, add 5mg / mL MTT20uL to each well, and ...
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