Methylene butyrolactone diterpene compound with hypoglycemic activity and preparation method and application thereof
A technology of methylene butyrolactone and compounds, which is applied in the field of medicine, can solve the problems of few research reports on chemical components, and achieve the effect of strong inhibitory activity
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Embodiment 1
[0029] The preparation of embodiment 1 methylene butyrolactone diterpene compound
[0030] (1) Get 8kg of Sanbaicao coarse powder, adopt supercritical CO 2 Extraction, extraction pressure 30MPa, extraction temperature 50°C, extraction time 2h, 95% ethanol as entrainer, the amount of entrainer added is 1 times the amount of medicinal materials, and the extract (about 110g) is obtained after vacuum concentration;
[0031] (2) Take the extract from step (1) and go to silica gel column chromatography (wet column loading, dry loading), and then use petroleum ether-ethyl acetate (volume ratio 50:1~0:100), ethyl acetate - Methanol (volume ratio 50: 1 ~ 0: 100) gradient elution to obtain 10 components (Fr.1 ~ Fr.10), according to thin-layer chromatography detection, take the Fr.4 part through the MCI column to remove the pigment (90 % ethanol elution), ODS medium pressure preparation, methanol-water (1:1 ~ 0:1) gradient elution to obtain Fr.4-1 ~ Fr.4-5, Fr.4-3 by semi-preparative HP...
Embodiment 2
[0035] Example 2 In vitro hypoglycemic test
[0036] 1. α-amylase inhibitory activity
[0037] Add 70 μL of PBS salt buffer solution (pH=6.9), 5 μL of 30 μg / mL α-amylase, and 2 μL of a certain concentration of methylenebutyrolactone diterpene sample to a 96-well plate sieve, and add 20 μL after incubating at 37°C for 10 minutes 10% starch substrate, react at 37°C for 20min, add 3μL I 2 -KI solution to stop the reaction, measure the absorbance value (A value) at 550nm; simultaneously set blank control (buffer solution only), negative control (buffer solution and enzyme solution only) and positive control (acarbose). All the above samples and blank reactions were repeated 3 times, and the average value was taken as the experimental data.
[0038] Inhibition rate (%) = [(A 酶 -A 酶空 )-(A 样 -A 样空 )] / (A 酶 -A 酶空 )×100%
[0039] Experimental results: The inhibitory rates of α-amylase at 6 concentrations of 1 μg / mL, 2 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL and 50 μg / mL of the compo...
Embodiment 3
[0044] The preparation of embodiment 3 capsules
[0045] Take the methylenebutyrolactone diterpene prepared in the above-mentioned Example 1 and add an appropriate amount of starch as a pharmaceutical excipient, mix thoroughly, put into capsules, and make capsules for oral use.
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