Diterpene compound, pharmaceutical composition containing same and preparation method and usage thereof
A compound and composition technology, applied in the field of medicine, can solve the problems of no compound activity report, no literature report, etc.
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Embodiment 1
[0020] Kunming mountain crabapple (10kg) with peeled roots was pulverized, 95% ethanol was reflux extracted three times (3×30L), each time for 5h, and concentrated under reduced pressure to obtain extract (575g). The extract was suspended in water (3L), extracted three times with ethyl acetate (3×3L), and concentrated under reduced pressure to obtain ethyl acetate extract (230g). The ethyl acetate extract was separated by silica gel column chromatography, petroleum ether-ethyl acetate-methanol gradient elution, the gradient was 20:9:1, 20:8:2, 20:7:3, 20:6:4 , 20:5:5, 20:4:6, get 6 components. Fraction 5 (85g) was separated by silica gel column chromatography, dichloromethane-acetone gradient elution, the gradient was 10:1, 8:1, 6:1, 4:1, 1:1, and 5 fractions were obtained . Fraction 4 (6 g) was purified by preparative HPLC with methanol-water isocratic elution with a gradient of 2:1 to obtain pure compound Ⅰ (45 mg).
[0021] Compound Ⅰ structure confirmation: amorphous ye...
Embodiment 2
[0025] The MTT method is simple, economical, fast, and free of radioactive contamination, and its detection results are in good agreement with the isotope incorporation method, which can be effectively applied to lymphocyte transformation. Therefore, the present invention adopts the MTT method, uses Balb / c mouse spleen lymphocytes, and targets at inhibiting the proliferation of mouse T and B lymphocytes, and uses different concentrations to study the activity of inhibiting cell proliferation of the compounds of the present invention.
[0026] 1. Animals and reagents
[0027] (1) Experimental animals: BALB / C mice, male, 18-20 g, purchased from Shanghai Experimental Animal Center.
[0028] (2) Experimental reagents: trypan blue solution, purchased from Invitrogen, USA; erythrocyte lysate, purchased from Beyontien Biological Reagent Company, Con A, purchased from Sigma, LPS, Sigma, USA.
[0029] 2. Experiment preparation
[0030] (1) Preparation of main reagents:
[0031] The ...
Embodiment 3
[0057] Preparation of injection preparations:
[0058] According to the method of Example 1, the compound I and the salt prepared by tartaric acid were firstly prepared, and water for injection was added as usual, finely filtered, potted and sterilized to make an injection.
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