Norlignan compounds and method of separating and verifying norlignan compounds from pouzolzia zeylanica var. microphylla
A technology of norelignan and multi-branch fog water kudzu, which is applied in the fields of organic chemistry, drug combination, antibacterial drugs, etc.
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Embodiment 1
[0044] The method for isolating the compound shown in formula (I) from Pueraria multibranches:
[0045] (1) Extraction: Weigh 10kg of Pueraria japonica, cut it into pieces, put it into an extraction tank, add 15 times the volume of 75% ethanol, and reflux extraction three times, each time for two hours, the extracts are combined and concentrated to obtain fine and crude extracts. things, spare.
[0046] (2) Liquid-liquid extraction and separation: disperse the extract with an appropriate amount of water, extract with petroleum ether, chloroform, and ethyl acetate in sequence until the color of each extract is light, and concentrate the ethyl acetate extract under reduced pressure to obtain a liquid extract.
[0047] (3) Silica gel column chromatographic separation: 21g of ethyl acetate extract was subjected to silica gel column (100~200 mesh, 1500g) chromatographic separation, and gradient elution was carried out with dichloromethane and methanol. The volume ratio of dichlorom...
Embodiment 2
[0062] Embodiment 2 antibacterial activity experiment
[0063] 1. Experimental materials
[0064] 1.1 Strains: Staphylococcus aureus strains were isolated and preserved by the Microbiology and Immunity Experiment Preparation Room of the School of Basic Science, Guangdong Pharmaceutical University.
[0065] 1.2 Reagent: the compound represented by formula (I) obtained in Example 1; dimethyl sulfoxide (Guangzhou Chemical Reagent Factory); Coptidis herb (Guangzhou Zhixin Pharmaceutical Co., Ltd.);
[0066] 1.3 Culture medium: nutrient agar bacterial dry powder medium, nutrient broth bacterial dry powder medium (Guangdong Huankai Microbial Technology Co., Ltd.);
[0067] 2. Method
[0068] 2.1 Preparation of samples
[0069] Accurately weigh 2 mg of the powder of the compound represented by formula (I) prepared in Example 1 into a sterilized EP tube, add 500 μL of sterilized 1% DMSO to dissolve it, and the concentration is 4 mg / mL. Store at 4°C for later use.
[0070] 2.2 P...
Embodiment 3
[0079] Example 3 Tumor activity experiment
[0080] 1. Experimental materials
[0081] 1.1 Drugs: the compound represented by formula (I) prepared in Example 1
[0082] 1.2 Cells: osteosarcoma cell line MG-63, breast cancer cell line SKBR-3
[0083] 1.3 Reagents: PRMI1640, fetal bovine serum, trypsin, DMSO, MTT, penicillin, streptomycin (Sigma); 96-well plate (Coring)
[0084] 2. Method
[0085] Osteosarcoma MG-63 cells and breast cancer SKBR-3 cells were separately cultured in 10% fetal bovine serum, 1×10 5 U / L penicillin, streptomycin in PRMI1640 culture medium, 37°C, 5% CO 2 Cultured in an incubator. Take the cells in the logarithmic growth phase, digest with 0.25% trypsin, pipette into a uniform single-cell suspension, count the cells, and dilute them with medium to 5×10 4 Inoculate cell / mL in a 96-well plate, 200 μL per well, 37°C, 5% CO 2 Incubate in an incubator for 24 h. The dosage of the experimental group was 3.13 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM...
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