New application of pregnane glucoside compound
A technology of pregnane glycosides and compounds, applied in the field of application in the preparation of immunosuppressive drugs and anti-inflammatory drugs
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Embodiment 1
[0014] Embodiment 1: Preparation of Epigynoide G
[0015] Get air-dried 15kg Laos florida Simao cane sample, pulverize and extract 3 times with methanol reflux, dissolve in water after concentrating under reduced pressure, then extract 3 times with ethyl acetate, concentrate the ethyl acetate layer, and weigh to obtain 1.3kg; The ethyl ester layer was roughly divided with macroporous adsorption resin D101, and eluted with aqueous methanol solutions of 40%, 60%, 80%, and 100% volume percentage respectively to obtain 4 parts (Fr.A, Fr.B, Fr. C, Fr. D). Fr.D (200g) was mixed with 1.5 times the amount of silica gel, mixed with silica gel for routine drying, packed into a column, and used chloroform-acetone (0:100-90:10) two-phase system as the mobile phase for gradient elution, according to TLC View and merge to get three parts Fr.D1-D3; Fr.D2 uses C 18 Reverse Phase C 18 Two fractions were obtained by column methanol-water gradient elution, Fr.D2a-D2b; Fr.D2a was eluted throug...
Embodiment 2
[0016] Embodiment 2: Immunosuppression detection test
[0017] (1) Preparation of spleen lymphocyte suspension
[0018] Take 18-22g healthy Balb / c mice to remove their eyeballs and bleed, and then kill them by taking off their necks, soak them in alcohol with a volume concentration of 75% for 3-5 minutes, take them out and place them in a sterile tray with the left side of the abdomen facing upwards, and put them in a safe place. In the cabinet, use sterilized tweezers to pick up the fur in the middle of the abdomen, make an incision from the lower end, use another set of instruments to cut open the layers of the abdominal wall, use the third set of instruments to remove the spleen, remove fat and connective tissue, and put it in PBS (Phosphate buffered saline) to wash away floating blood. Then move the spleen tissue to a plate filled with RPMI1640 incomplete culture medium, cut it into small pieces with scissors, use a sterile syringe core to gently grind the spleen in a 200...
Embodiment 3
[0046] Embodiment 3: in vitro anti-inflammatory activity test
[0047] (1) Preparation of test-related reagents
[0048] Preparation of sample dilution working solution: The concentrated sample dilution solution was taken out from the -4°C refrigerator, thawed slowly at room temperature, and diluted according to the amount required for the test with double distilled water at a ratio of 1:5.
[0049] Preparation of reagent dilution working solution: take the concentrated reagent solution out of the -4°C refrigerator, thaw slowly at room temperature, take part according to the amount required for the test, and dilute it with double distilled water at a ratio of 1:5.
[0050] Preparation of cell lysis working solution: The concentrated cell lysate was taken out from the refrigerator at -4°C, thawed slowly at room temperature, and diluted with double distilled water at a ratio of 1:2 according to the amount required for the test.
[0051] Preparation of washing working solution: ...
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