Low-molecular-weight fucosylated chondroitin sulfate, preparation method thereof and application of low-molecular-weight fucosylated chondroitin sulfate to preparation of medicine for resisting Trousseau syndrome
A chondroitin sulfate and fucosification technology, which is applied in the application field of medicine, can solve the problem that no relevant reports have been reported on the research and application of fucosylated chondroitin sulfate, the pH of the reaction system is unstable, and the content of fucose sulfate is reduced. To achieve the effect of inhibiting lung metastasis, relieving Trusseau syndrome, and easy to prepare in large quantities
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Embodiment 1
[0048] Example 1: Preparation of low molecular weight fucosylated chondroitin sulfate
[0049] 1. Take 100 g of North Atlantic dried sea cucumber, homogenize it, defat it, add 0.1 mol / L NaOH solution, stir and react at 50 ℃ for 3 h;
[0050] After cooling at room temperature, add hydrochloric acid to neutralize, add 500 mg papain, 5 mmol / L Na 2 EDTA solution and 5 mmol / L cysteine solution were stirred and reacted at 50 °C for 12 h;
[0051] Combine the extracts, add 3 times the volume of 95% ethanol to precipitate, centrifuge, wash the precipitate 3 times with 95% ethanol and absolute ethanol, and dry at 60 °C;
[0052] Take the dried sample, add appropriate amount of water to dissolve, add 0.1 mol / L KOH solution to adjust the pH to 9, add 30% H 2 o 2 , decolorize at 30°C for 2 h;
[0053] After the reaction solution was cooled, hydrochloric acid was added to adjust the pH to 1-2, and the precipitate was removed by centrifugation;
[0054] Add 95% ethanol until the alco...
Embodiment 2
[0062] Example 2: The effect of low molecular weight fucosylated chondroitin sulfate against Trusso syndrome
[0063] 1. Passage Lewis lung cancer cells in vivo: Take Lewis lung cancer cells in the logarithmic growth phase, digest with 0.25% trypsin, and adjust to 1×10 with normal saline 7 cells / ml of single-cell suspension, and inoculated 0.2 ml subcutaneously in the right axillary of mice. 10 days after inoculation, take the Lewis lung cancer solid tumor under sterile conditions, make a single cell suspension according to the ratio of tumor block (g): normal saline (ml) 1:3, take 0.2 ml (containing tumor cells 2×10 6 ) were inoculated subcutaneously in the right axilla of C57BL / 6J mice, and passaged ten times in vivo.
[0064] 2. Replication and experiment of animal model: take the Lewis lung cancer solid tumor that has been passaged ten times in vivo under sterile conditions, make a single-cell suspension according to the ratio of tumor mass (g): normal saline (ml) at 1:3...
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