Different terpenoid intermolecular adducts in Pseudotsuga forrestii Craib and preparation method and pharmaceutical application thereof
An intermolecular and adduct technology, applied in the separation/purification of carboxylic acid compounds, medical preparations containing active ingredients, drug combinations, etc., to achieve significant ACL inhibitory activity
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Embodiment 1
[0024] Embodiment 1: prepare compound described in the present invention
[0025] (1) After drying and pulverizing the branches and leaves of Douglas chinensis collected from Dali, Yunnan (first batch: collected in October 2017, dry weight 13kg), they were extracted 7 times with 90% methanol (6L) solution at room temperature, each time 24 hours;
[0026] (2) Combine the extracts obtained in step (1), concentrate under reduced pressure to remove methanol to obtain 2.46kg (semi-dry) extract, disperse the extract with 2L of water, and use equal volumes of petroleum ether, ethyl acetate and n-butyl Alcohol was extracted three times respectively, and each extract was concentrated under reduced pressure to obtain petroleum ether part, ethyl acetate part, n-butanol part and water 4 components respectively, wherein, ethyl acetate part crude extract (409g) was passed through silica gel column Chromatography (washing with petroleum ether-ethyl acetate gradient, 30:1→0:1, v / v) separated...
example 2
[0042] Example 2: ACL inhibitory activity test of the compound prepared by the present invention
[0043] Experimental method: In this experiment, ACL can catalyze the conversion of citric acid into acetyl-CoA, and then produce the precursor molecule of fatty acid synthesis - malonyl-CoA. This reaction is accompanied by the consumption of ATP, so ADP-Glo and kinase can be used The detection kit detects the change of ATP, and the inhibitory effect of the indirect reaction compound on the ACL enzyme activity; in this embodiment, the percentage inhibition rate of the ACL enzyme activity was investigated when the concentration of the compound selected for the primary screening was 20 μg / ml, and the test results It shows that the above compounds all have enzyme activity inhibitory effect;
[0044] Further determination of IC 50 Value: the sample is dissolved in DMSO to make a suitable concentration before use, 3-fold dilution, 7 dilutions, triplicate wells, take 2μL sample solut...
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