Application of mannoglucuronic acid oligosaccharides and derivatives thereof in the preparation of medicines for treating and/or preventing hcmv
A technology of glucuronic acid and uronic acid oligosaccharides, which is applied in the application field of preparing medicines for treating and/or preventing HCMV, and achieves the effect of less toxic and side effects
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Embodiment 1
[0116] The preparation of embodiment 1 mannoglucuronic acid oligosaccharides
[0117] Extract the dried kelp with 30 times the mass of distilled water at 100°C for 3 hours, filter the extract, concentrate, add ethanol to a final concentration of 75% to precipitate, let it stand for 12 hours, collect the precipitate, sink to the point of vacuum drying to obtain laminarin . Dissolve the laminarin sample in a sulfuric acid solution with a mass concentration of 4% (the ratio of solid to liquid is 60mg / mL), heat and reflux for 5 hours, neutralize with barium hydroxide to PH=6-7, centrifuge, and concentrate the supernatant to the original One-fifth of the volume, the concentrated solution is subjected to active carbon column chromatography, firstly equilibrated with distilled water, and then eluted with a gradient of 50%-90% ethanol, and the 50%-90% ethanol eluate is concentrated to one-fifth of the original volume 1. Ethanol was evaporated, and directly applied to Bio-gel P4 colum...
Embodiment 2
[0124] The preparation of embodiment 2 sulfated mannoglucuronide oligosaccharides
[0125] The mannoglucuronide oligosaccharide monomers GM2, GM4, GM6 (mannose glucuronide disaccharide, tetrasaccharide, hexasaccharide) obtained in Example 1 were dissolved in DMF (0.05g / ml) under nitrogen protection conditions, Then add sulfur trioxide pyridinium salts of different quality, and stir at room temperature for 24 hours. The reaction solution was poured into 4 times ice-distilled water, neutralized, and desalted with Sephadex G10. The eluate was concentrated, separated and purified on Bio-gel P4 to obtain GlcAMan (SO 3 h) 1-3 , GlcAMan (SO 3 h) 3-5 , GlcA 2 man 2 (SO 3 h) 8-11 , GlcA 2 man 2 (SO 3 h) 5-9 , GlcA 2 man 2 (SO 3 h) 1-5 , GlcA 3 man 3 (SO 3 h) 4-10 , GlcA 3 man 3 (SO 3 h) 1-6 , GlcA 3 man 3 (SO 3 h) 8-15 .
Embodiment 3
[0126]Example 3 Effects of Mannoglucuronide Oligosaccharides and Derivatives Treated Alone on the Proliferation Activity of WI-38 Cells
[0127] The cell viability was detected by the MTT method as follows: PD30 human embryonic lung fibroblast WI-38 cells were seeded in 96-well cell culture plates at 3000 cells per well, and after 24 hours of culture, different concentrations of mannoglucuronide oligosaccharides were added. Three parallel wells were set up for each concentration, and a blank control group without drug and a cell-free lysis control group were set up. After culturing for 48 hours, add 20 μl MTT (5 mg / ml, prepared with serum-free DMEM culture solution) to each well, at 37°C, 5% CO 2 Incubate under the conditions for 4 hours, carefully suck up the liquid in the wells, add 150 μl DMSO to each well, and shake gently on the shaker for 10 minutes to fully dissolve the crystals, then measure the light absorbance at 570nm, and calculate the relative cell viability. The...
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