Naphthoquinone dimer and its use in the preparation of anti-complement drugs
A naphthoquinone dimer tiger and anti-complement technology, which can be applied in the fields of drug combination, quinone separation/purification, organic chemistry, etc., and can solve problems such as no reports of anti-complement active compounds
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Embodiment 1
[0017] The preparation of embodiment 1 polydatin C
[0018] Polygonum cuspidatum root coarse powder 16kg, extract 3 times with 5 times of amount 95% ethanol 50 ℃, each 2 hours. The solvent was recovered under reduced pressure to obtain 950 g of extract, which was suspended in distilled water and extracted with petroleum ether, ethyl acetate and n-butanol to obtain 400 g of ethyl acetate extract. Take 200 g of the ethyl acetate extract and go through silica gel column chromatography, eluting with dichloromethane, dichloromethane-methanol, methanol gradient, and the obtained fractions are subjected to repeated silica gel column chromatography and SephadexLH-20 purification with different eluents, and the compound is isolated Polygonadin C.
[0019] The specific steps are as follows: the obtained fraction was eluted with dichloromethane-methanol (20:1), eluted with petroleum ether-acetone gradient, and purified by SephadexLH-20 to obtain the compound (2 mg), which was identified...
Embodiment 2
[0020] Example 2 Anti-complement classical pathway test in vitro
[0021] Take 0.1ml of complement (guinea pig serum), add BBS buffer to make a 1:5 solution, and double-dilute with BBS to 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:640 solution. Take 0.1ml each of 1:1000 hemolysin and 2% SRBC, and dissolve 0.2ml of each concentration of complement in 0.2ml BBS, mix well, put it in a low-temperature high-speed centrifuge after 30 minutes in a 37°C water bath, and centrifuge at 5000rpm and 4°C 10min. 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 (0.1ml 2% SRBC dissolved in 0.5ml triple distilled water) was set up. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Taking the dilution of complement as the X-axis, the percentage of hemolysis caused by each dilution of comple...
Embodiment 3
[0023] Example 3 Anti-complement alternative pathway test in vitro
[0024] Take 0.2ml of complement (human serum), add AP diluent to prepare a 1:5 dilution solution, and double-dilute to 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:640 solution. Take 0.15ml of complement of each concentration, 0.15ml of AP diluent and 0.20ml of 0.5% RE, mix well, place in a low-temperature high-speed centrifuge after 30min in a 37°C water bath, and centrifuge at 5000rpm and 4°C for 10min. 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. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Taking the dilution of complement as the X-axis, the percentage of hemolysis caused by each dilution of complement is plotted as the Y-ax...
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