Compound with anti-tumor activity and its preparing method
A compound and extract technology, applied in the field of novel triterpenoids and their preparation, can solve the problem of no anti-tumor activity and the like
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Embodiment 1
[0052] Example 1 Extraction of 7β, 24β, 25-trihydroxy-9,19-cycloartin-1-en-3-one 25-O-β-D-glucopyranoside
[0053] A. After crushing 1kg of bitter horse bean, put it into the extraction tank
[0054] B. 8 L of 95% ethanol was refluxed and extracted 3 times, each time for 2 hours, and the ethanol was recovered to obtain 103 g of ethanol extract.
[0055] C. Dissolve the ethanol extract in 1L of water, and extract 3 times with an equal volume of chloroform to obtain 33.7 g of chloroform extract.
[0056] D. The chloroform extract was subjected to silica gel column chromatography (wet packing, dry loading), the ratio of the sample volume to the total amount of silica gel was 1:5, and after elution with chloroform:methanol (100:2) for 3 retention volumes, Recrystallization gave 7β, 24β, 25-trihydroxy-9,19-cycloartin-1-en-3-one 25-O-β-D-glucopyranoside (147 mg).
Embodiment 2
[0057] Example 2 Extraction of 7β, 24β, 25-trihydroxy-9, 19-cycloartin-1-en-3-one 25-O-β-D-glucopyranoside
[0058] a After crushing 1kg of bitter horse bean, put it into the extraction tank
[0059] b Soak and extract in 10L methanol for 3 times, each time for 12 hours, recover methanol, and obtain 116g of methanol extract.
[0060] c Dissolve the methanol extract in 800 mL of water, and extract twice with an equal volume of dichloromethane to obtain 31.9 g of dichloromethane extract.
[0061] d The dichloromethane extract was subjected to alumina column chromatography (wet column loading, dry loading), the sample volume to alumina weight ratio was 1:10, and 4 samples were eluted with dichloromethane:methanol (100:3). After volume retention, recrystallization gave 7β,24β,25-trihydroxy-9,19-cycloaltin-1-en-3-one 25-O-β-D-glucopyranoside (132 mg).
Embodiment 3
[0062] Example 3 Anti-tumor effect of 7β, 24β, 25-trihydroxy-9, 19-cycloartin-1-en-3-one 25-O-β-D-glucopyranoside in vitro
[0063] Take the cells in the logarithmic phase 3×10 per well 4 Inoculated on a 96-well plate, discarded the supernatant after 12 hours, and administered according to the following groups: Tumor cells were divided into groups without treatment and groups with treatment (concentration 0-100 μM), each group had 6 replicate wells, cultured for 24 hours, discarded Add 50 μl of culture medium with MTT (tetrazolium salt, Sigma product) to the supernatant and incubate for 4 hours (0.5 mg / mL), add 100 μl DMSO (dimethyl sulfoxide), shake for 1 hour, and measure at 570 nm on a microplate reader OD value. The normal cell line L02 was used as a control. Repeat 3 times.
[0064] The results showed that as the drug concentration increased, the cell proliferation activity decreased respectively compared with the corresponding control group without drug addition, indi...
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