Application of diphenol compounds in preparation of anti-complement medicaments
A kind of compound, the technology of hydroquinone, applied in the new use field in the preparation of anti-complement medicine
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Embodiment 1
[0022] Example 1 Preparation of n-butanol extract to obtain hydroquinone compounds
[0023] Take 15kg of dried and mature Tsao Kuo fruit, cold-soak 50L×10 times with ethanol at room temperature, combine the extracts and concentrate until there is no alcohol smell, dilute the extracts with water to 2L, and extract 2L× each with petroleum ether, ethyl acetate, and n-butanol in sequence 3 times, the combined n-butanol extracts were concentrated to dryness to obtain 86.2 g of n-butanol extracts. The n-butanol part was treated with AB-8 macroporous adsorption resin, and the 50% ethanol elution part (21.3g) was subjected to silica gel column chromatography, and was eluted with petroleum ether (60-90°C), petroleum ether-acetone, and acetone gradient. Specific steps are as follows:
[0024] 1. The resulting fractions were eluted with petroleum ether-acetone (3:1), followed by repeated silica gel column chromatography with chloroform-acetone (5:1) to obtain compounds (1) (10 mg), (3) ...
Embodiment 2
[0026] Example 2 Anti-complement classical pathway test in vitro
[0027]Take 0.1ml of complement (guinea pig serum), add BBS to prepare a 1:5 solution, and dilute it to 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:320 with BBS 640 solution. Dissolve 1:1000 hemolysin, 0.1ml of each concentration of complement and 2% SRBC in 0.3ml BBS, mix well, put 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. A full hemolysis group (0.1ml 2% SRBC dissolved in 0.5ml triple distilled water) was also 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. Taking the dilution of complement as the X-axis, the percentage of hemolysis caused by each dilution of complement is plotted as the Y-axis. ...
Embodiment 3
[0028] Example 3 Anti-complement alternative pathway test in vitro
[0029] 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 30 minutes in a 37°C water bath, and centrifuge at 5000rpm and 4°C for 10 minutes. 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. Taking the dilution of complement as the X-axis, the percentage of hemolysis caused by each dilution of compl...
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