A kind of sea cucumber alkaloid and its preparation method and application
An alkaloid and sea cucumber technology, which is applied in the directions of drug combination, organic chemistry, anti-tumor drugs, etc., can solve the problem that the anti-tumor activity of sea cucumber alkaloid compounds has not been reported, and achieve the effect of simple preparation method and low cost.
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Embodiment 1
[0032] (1) After taking out the viscera of 100kg of fresh sea cucumbers, put them in boiling water and cook for 20-30 minutes, and 100kg of sea cucumbers will produce about 50kg of sea cucumber boiling liquid;
[0033] (2) Get 20L of sea cucumber boiled liquid, adjust the pH value to 10, and centrifuge at 8000r / min for 40min to obtain the supernatant, and pass the obtained supernatant in batches through AB-8 type macroporous resin columns (5L each time), the flow rate 2BV / h, after the adsorption is saturated, rinse with 2L of deionized water, and then rinse with 2L of 20% ethanol solution;
[0034] (3) Elute the above-mentioned macroporous resin column with 6L of 70% ethanol solution, collect the eluent, concentrate under reduced pressure to extractum, and obtain sea cucumber extract;
[0035] (4) Redissolve the above-mentioned sea cucumber extract with 500mL of deionized water, centrifuge at 8000r / min for 40min to take the supernatant, extract and degrease the supernatant wit...
Embodiment 2
[0045] The in vitro antitumor test was carried out on the 6-OH erythracine prepared in Example 1, which showed that it had a moderately strong antitumor effect. The cell lines used in the test are internationally accepted tumor cell lines, namely: lung cancer cell A-549, NCI-H1299, liver cancer cell HepG2, prostate cancer cell PC-3 and breast cancer cell MCF-7.
[0046] 6-OH erythrine was formulated with DMSO at a standard concentration of 50 μg / mL, filtered through a 0.45 μL microporous membrane, and diluted sequentially to 25, 12.5, 6.25 and 3.125 μg / mL before use and added to the cells to be tested. The determination of antitumor activity adopts MTT method:
[0047]a) Collect tumor cells in the logarithmic growth phase at 1×10 5 / mL concentration inoculated in 96-well plate, 100 μl / well, at 37°C, 5% CO 2 Cultivate in the incubator for 24 hours to make the cells adhere to the wall;
[0048] b) Aspirate the supernatant, add the maintenance solution containing the sample so...
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