Saururus chinensis extract as well as preparation method and application thereof
A kind of Sanbaicao extract and extract technology, applied in the direction of medical formula, plant raw materials, active ingredients of heterocyclic compounds, etc., can solve the problem that there is no compound to inhibit virus cleavage and replication, and there is no anti-EBV virus cleavage and replication of Sanbaicao extract Activity and other issues, to achieve the effect of low toxicity and good antiviral effect
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Embodiment 1
[0033] The method for preparing compound formula (I) and formula (II) from three white grass (Saururus chinensis):
[0034] The whole plant of Saururus chinensis (5kg), sun-dried, crushed, extracted three times with 95% ethanol under reflux, each time for 2 hours, combined the concentrated solution to obtain the crude extract (296g), and extracted with petroleum ether and ethyl acetate respectively Ethyl acetate and n-butanol extraction, ethyl acetate (68g) part was directly subjected to silica gel column chromatography (1500g, 200-300 mesh, 7.0×100cm), and petroleum ether-ethyl acetate (100:0, 95:5, 90: 10, 80:20, 70:30, 60:40, V / V) gradient elution, each 500mL was collected, and the same fraction was combined for TLC detection to obtain a total of nine fractions Fr. A-I. Part of Fr. C (15g) was eluted with silica gel column chromatography (250g, 200-300 mesh, 4.5×60cm) with petroleum ether-ethyl acetate (90:10, V / V) to obtain four fractions Fr. .C1-4, Fr.C1 (2g) was elute...
Embodiment 2
[0065] Example 2 Pharmacodynamic related experiments of the 10 compounds obtained in Example 1
[0066] Determination of inhibitory activity of compounds 1-10 of the present invention on EBV cleavage and replication.
[0067] (1) Cell culture. P3HR-1 cells (primary effusion lymphoma cell line containing latently infected EBV) were cultured in vitro. Use RPMI1640 medium containing 10% fetal bovine serum, streptomycin (100 μg / ml) and penicillin (100 units / ml) for routine maintenance and passage at 37°C and 5% carbon dioxide concentration.
[0068] (2) Drug intervention. Adjust the density of P3HR-1 cells in the logarithmic growth phase to 3×10 5 cells / ml, use 20ng / ml Tetradecanoylphorbol acetate (TPA) and sodium butyrate (0.3 mM) to induce P3HR-1 cells to enter the lytic replication phase. The compounds to be tested were formulated into drug solutions with different concentrations using DMSO. After P3HR-1 cells were treated with TPA for 3 hours, the cells were treated with ...
Embodiment 3
[0073] Embodiment 3 Compounds 1 to 10 of the present invention are tested for host cell toxicity
[0074] (1) Cell culture. P3HR-1 cells (primary effusion lymphoma cell line containing latently infected EBV) were cultured in vitro. Use RPMI1640 medium containing 10% fetal bovine serum, 400ug / ml G418, 100ng / ml doxycycline, at 37 ℃, 5% carbon dioxide concentration conditions for routine maintenance and passage.
[0075] (2) Test method. Adjust the density of P3HR-1 cells in the logarithmic growth phase to 3×10 5 cells / ml, the cells were treated with different concentrations of compounds, and three parallel wells were set up for each concentration. After 2 days, trypan blue staining was used to count the number of living cells under a light microscope.
[0076] (3) Result processing. According to the formula: relative toxicity = 1-live cell compound+ / live cell compound_ Calculate the relative toxicity and median lethal dose (CC) of each compound at different concentrations ...
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