Fuc3S4S substituted oligo-glycosaminoglycan and preparation method thereof
A technology of glycosaminoglycan and polysaccharide amine, which is applied in the field of Fuc3S4S substituted oligosaccharide aminoglycan and its preparation, and can solve the problems of reducing the bleeding tendency of anticoagulant active ingredients and the like
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Embodiment 1
[0159] Extraction and purification of polysaccharide fractions of sea cucumber containing Fuc3S4S substituted glycosaminoglycans (HSG-1, HSG-1, HSG-3)
[0160] 1.1 Materials
[0161] Raw materials: dry rough sea cucumber Holothuria scabra, purchased from the market.
[0162] Reagent: papain, 8×10 5 U / g, Nanning Pangbo Bioengineering Co., Ltd., Guangxi. NaOH, KCOCH 3 、H 2 o 2 , ethanol and other reagents are commercially available analytical reagents.
[0163] 1.2 Extraction and purification
[0164] Extraction: Take 1000g of dry rough sea cucumber, mechanically slice and crush it, put it in a round bottom flask, add 10L of pure water to soak overnight, after the water bath is heated to 50°C, add 5g of papain and mix well, stir and enzymatically hydrolyze at 50°C for 6h . Subsequently, solid sodium hydroxide was added to a concentration of about 0.5M, and alkaline hydrolysis treatment was carried out at 60° C. for 2 h. Thereafter, adjust the pH value to 6-7 with 6N hyd...
Embodiment 2
[0180] Preparation of Fuc3S4S-substituted oligoglycosaminoglycan mixture (dHSG-1) by depolymerizing the polysaccharide fraction of sea cucumber by peroxidative depolymerization
[0181] 2.1 Materials
[0182] HSG-2: Example 1 extracts and purifies the polysaccharide fraction containing Fuc3S4S substituted glycosaminoglycans from the dried rough sea cucumber Holothuria scabra. Mw, ~63.5kD; PDI, ~1.06.
[0183] Reagents: NaCl, CH 3 COONa, NaOH, H 2 o 2 , Cu(CH 3 COO) 2 All reagents are commercially available analytical grade reagents.
[0184] 2.2 Preparation
[0185]Take HSG-2100.3mg in Example 1, place in a 10mL reaction test tube, add 3mL water to dissolve, add 87.2mg NaCl to make the concentration 0.5M, add 204.1mg sodium acetate trihydrate to make the concentration of sodium acetate 0.5M, add Copper acetate 2.4mg to make the concentration 4.01nM, add 0.4mL10% H 2 o 2 . The pH of the reaction solution was adjusted to about 7.5 with 0.1M NaOH, and depolymerization ...
Embodiment 3
[0203] Preparation of Fuc3S4S-substituted oligoglycosaminoglycan mixture (dHSG-2) by depolymerization of sea cucumber polysaccharides by deacylation, deamination and depolymerization
[0204] 3.1 Materials
[0205] HSG-3: Extracted and purified polysaccharide components containing Fuc3S4S substituted glycosaminoglycans from the dried rough sea cucumber Holothuria scabra in Example 1. Mw, ~64.2kD; PDI, ~1.09.
[0206] Reagents: Hydrazine hydrate, hydrazine sulfate, absolute ethanol, sodium borohydride, etc. are commercially available analytical reagents.
[0207] 3.2 Preparation
[0208] (1) Preparation of Partially Deacetylated Products : Take the rough sea cucumber polysaccharide component HSG-360mg containing Fuc3S4S substituted glycosaminoglycans described in Example 1 and place it in a reaction tube, add 14.5mg of catalyst hydrazine sulfate, then add hydrazine hydrate 1.45mL, under nitrogen protection, under 250rpm rotating speed Stir and heat the reaction at 70°C for...
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