Low-molecular sea cucumber glycosaminoglycan and application thereof
A glycosaminoglycan and low-molecular-weight technology, which can be used in medical preparations containing active ingredients, organic active ingredients, drug combinations, etc., can solve problems such as unsatisfactory pharmacological activity, and achieve anti-tumor or anti-tumor metastasis learning and memory capabilities. , regulate platelet activity, improve the effect of learning and memory ability
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Embodiment 1
[0029] Embodiment 1: Preparation of low molecular weight sea cucumber glycosaminoglycan (LHG)
[0030] This embodiment is the preparation of LHG of the present invention, and the preparation process includes the following steps:
[0031] Step 1. Extraction and purification of sea cucumber glycosaminoglycan (HG):
[0032] Weigh 10Kg of commercially available dried sea cucumbers, soak them in 20L of water overnight, and mince them. Transfer the sea cucumber homogenate into a 100L reactor, add 50L of water, 0.8Kg of sodium carbonate and 1.6Kg of sodium chloride, heat to 50°C, and keep warm. Add 0.2Kg of alkaline protease 2709 for 6 hours of enzymatic hydrolysis. After the enzymatic hydrolysis, remove the gravel by gauze filtration, and transfer the filtrate to a LANXESS S5428 resin kettle with a wet weight of 5Kg, and continue to stir and adsorb for 3 hours. Collect the resin by filtration, rinse the resin three times with water, rinse the resin with 15 L of 4% sodium chloride...
Embodiment 2
[0041] Embodiment 2: Physicochemical properties and biological characteristic analysis of LHG
[0042] This example is an analysis experiment of the physicochemical properties and biological characteristics of different batches of LHG prepared in Example 1.
[0043] 1. Experimental materials:
[0044] Compounds to be tested: each batch of LHG from Example 1 above;
[0045] 2. Experimental method:
[0046] (1) Weight-average molecular weight detection: adopt GPC-HPLC method, the specific conditions are: a TSKgel G2000SW (7.8mm×30cm, 5μm) and a TSKgel G3000SW (7.8mm×30cm, 5μm) are connected in series for the chromatographic column, and the detector is Refractive index detector (RID), the weight average molecular weight calculation standard is the weight average molecular weight calibrator RS of USP commercial low molecular weight heparin, the analysis software includes HPLC workstation and Agilent's GPC software; SEC-HPLC analysis and detection combined with GPC software Calc...
Embodiment 3
[0067] Embodiment 3: Structural characteristic analysis of LHG main chain disaccharide unit
[0068] This example is an analysis experiment on the structural characteristics of the main chain disaccharide unit of the LHG prepared in Example 1.
[0069] According to the literature, natural HG is composed of glucuronic acid (GlcA) and N-acetylgalactosamine (GalNAc) as the main chain of chondroitin sulfate (CS) repeating disaccharide units, and sulfated polyfucose (Fuc ) is a type of glycosaminoglycan (GAG) composed of branched chains. LHG is derived from natural HG, and its basic characteristics are similar, but there are various forms on the disaccharide repeating unit (dp2) of the main chain due to factors such as the degree and position of sulfate group or acetyl group substitution. Due to the existence of fucose branched chains, chondroitin sulfate enzyme cannot degrade the main chain of chondroitin sulfate of LHG, and thus cannot complete the disaccharide analysis of the m...
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