A kind of pharmaceutical composition, its preparation method and application
A composition and immune drug technology, applied in the field of pharmacy, can solve the problems of no immunosuppressive activity, etc., and achieve the effect of remarkable drug effect, simple preparation process, and less toxic and side effects
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Embodiment 1
[0033] The preparation of embodiment 1 pharmaceutical composition of the present invention
[0034] The extraction method of the southern yew extract comprises the following steps:
[0035] Take the southern yew root, add 10 times the amount of water and decoct three times, 2 hours each time, combine the decoction liquid, filter, and concentrate the filtrate under reduced pressure to 1-2g of the original drug per 1ml, and precipitate with 95% ethanol to the alcohol content up to 60%, refrigerated for 24 hours, filtered, the filtrate was taken to recover ethanol, refrigerated for 24 hours, filtered, the filtrate was passed through a macroporous resin column, washed with water until nearly colorless, eluted with 75% ethanol, and the eluate was collected. Filter and concentrate.
[0036] The extracted southern yew root extract is mixed with polyethylene glycol to make capsule medicine.
experiment example 2
[0037] Experimental example 2 in vitro verification
[0038] The experiment was divided into blank control group (only adding cell fluid), Con A group, LPS group, administration A group (only adding test drug), administration B group (adding Con A and test drug), administration C group (plus LPS and test drug).
[0039] (1) The mouse spleen was taken out, crushed, sieved, and washed 3 times with RPMI 1640 culture solution.
[0040] (2) Red blood cells were lysed with erythrocyte lysate, and centrifuged at 1000 rpm for 5 minutes. After centrifugation, resuspend in PBS and count the cells.
[0041] (3) Inoculate cells into 96-well cell culture plate, 1×10 6 cells / well.
[0042](4) For blank control group, Con A group, and LPS group, only 100 μL of culture solution was added. 100 μL of the RPMI1640 culture solution of the test drug was added to groups A, B, and C administered. After 1h, ConA (final concentration: 5 μg / mL) was added to Con A group and administration B group,...
experiment example 3
[0049] Experimental example 3 in vivo verification
[0050] Dinitrofluorobenzene is a hapten. Apply it to the abdominal skin and combine with skin protein to form a complete antigen to sensitize the skin. It will be strengthened once after 24 hours, and it will be applied to the ear on the 5th day of sensitization, causing delay. Hair allergy. The purpose of the experiment is to observe the inhibitory effect of the compound on allergic dermatitis in mice, and preliminarily evaluate its immunosuppressive activity.
[0051] Experimental method: ICR mice were selected and randomly divided into normal control group, model control group and drug treatment group, with 10 mice in each group. The test extracts were administered daily. Abdominal depilation, apply 1% DNFB evenly on the abdomen for sensitization, strengthen once 24 hours later in the same way, on the 5th day of sensitization, apply 1% DNFB evenly to the right ear of the mouse to attack, kill the mouse 24 hours later, c...
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