Uses of lignan compound (7S,8R)-dihydrodehydrodiconiferyl alcohol in preparation of anti-complement drugs
A technology of lignans and compounds, applied in the field of medicine, can solve the problems that have not yet been seen
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Embodiment 1
[0013] Example 1 Preparation of (7S, 8R)-dihydrodehydrodisconiferyl alcohol
[0014] Take 25kg of dry roots of Sesame chinensis, crush them, and reflux extract with 95% ethanol for 3 times (50L×3), each time for 2h, combine the extracts and concentrate to obtain 0.95kg of extract, add water (4L) to suspend, and add an equal volume of petroleum Extracted 5 times with ether, ethyl acetate and n-butanol, combined the extracts and concentrated to dryness to obtain 200 g of ethyl acetate extract. The ethyl acetate extraction part was separated by silica gel (200-300 mesh) column chromatography, and sequentially eluted with dichloromethane-methanol (50:1-0:1) gradient to obtain 11 fractions (Fr.1-11) , wherein the fraction Fr.3 (32g) was subjected to silica gel column chromatography (dichloromethane-methanol, 30:1, 20:1, 10:1, 5:1, 3:1, 1:1), MPLC (methanol - water, 20:80-80:20 gradient elution) and sephadexLH-20 column chromatography and other means to purify, isolate and obtain c...
Embodiment 2
[0016] Example 2 Anti-complement classical pathway activation test in vitro
[0017] Take 0.04ml of complement (guinea pig serum), add barbiturate buffer solution (BBS) to prepare a 1:10 solution, and double-dilute with BBS to 1:20, 1:40, 1:80, 1:160, 1:10 320, 1:640 and 1:1280 solutions. Take 1:1000 hemolysin, 0.1ml of 2% sheep red blood cells (SRBC) and 0.2ml of complement of each concentration, dissolve them in 0.2ml of BBS, mix well, put them in a low-temperature high-speed centrifuge at 4000rpm and 4℃ Centrifuge for 5min. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure its absorbance at 405nm. At the same time, a complete hemolysis group (0.1ml 2% SRBC, 0.1ml hemolysin dissolved in 0.4ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Plot the dilution of complement on the X-axis...
Embodiment 3
[0018] Example 3 Anti-complement Alternative Pathway Activation Test in Vitro
[0019] Take 0.2ml of complement (human serum), add AP diluent (barbital buffer, pH=7.4, containing 5mMMg 2+ ,8mMEGTA) was prepared as a 1:5 solution, and double-diluted into 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:640 solutions. Take 0.15ml of complement of each concentration, 0.15ml of AP diluent and 0.20ml of 0.5% rabbit erythrocytes (RE), mix them evenly, put them in a low-temperature high-speed centrifuge at 37°C for 30min, and centrifuge at 4000rpm and 4°C for 5min. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405nm. At the same time, a complete hemolysis group (0.20ml 0.5% RE dissolved in 0.3ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Plot the dilution of complement on t...
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