Depolymerization glycosaminoglycan extracted from sea cucumber composition and its preparation method and application

A technology of glycosaminoglycans and compositions, which is applied in the field of prevention and treatment of cardiovascular and cerebrovascular diseases, and can solve the problem that the safety, effectiveness and quality of low-depolymerized sea cucumber glycosaminoglycans cannot meet the pharmaceutical standards , Difficult to remove fucose from sea cucumbers, etc., to achieve anticoagulant mechanism and excellent safety, prevention and treatment of coagulation and thrombosis diseases, and low price

Active Publication Date: 2007-10-24
XIAN XINTONG PHARM RES CO LTD
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  • Description
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Problems solved by technology

Chinese patent application CN1440985A discloses a method for preparing polysaccharides that can be extracted from sea cucumbers. The polysaccharides in the prepared sea cucumber polysaccharides have a large molecular weight, and it is difficult to remove the sea cucumber fucose mixed in the glycosaminoglycans, and the quality is difficult to achieve. pharmaceutical standard
For this reason, Chinese patent application CN1579415A and people such as Hideki Nagase (can refer to Thromb Res; 2003,112 (1-2): 83-91; Kidney Int., 2003, 63 (4): 1548-55; Thromb Haemost., 1997, 77(6): 1148-53; Thromb Haemost., 1997, 77(2): 399-402; Thromb Res., 1996, 15; 84(2): 111-20; Thromb Res., 1996, 83 (3): 253-64; J Biochem (Tokyo)., 1996, 119(1): 63-9; Blood., 1995, 85(6): 1527-34, the full texts of which are incorporated herein by reference) respectively By further depolymerizing glycosaminoglycans from sea cucumbers to obtain a composition of depolymerized sea cucumber glycosaminoglycans with a molecular weight distribution in the range of 10,000 Da to 15,000 Da, such compositions can be used to replace drugs such as heparin in some in vivo or in vitro experiments However, there is no sufficient experimental evidence in these documents to show the medicinal safety of these compositions, and there is no suggestion that the depolymerized sea cucumber glycosaminoglycan composition with a lower average molecular weight can be used as an anticoagulant drug for prevention and treatment. safety and effectiveness
In addition, the sea cucumber glycosaminoglycan drugs with large molecular weight that are currently being worked on still have side effects such as easy platelet aggregation that need to be resolved

Method used

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  • Depolymerization glycosaminoglycan extracted from sea cucumber composition and its preparation method and application
  • Depolymerization glycosaminoglycan extracted from sea cucumber composition and its preparation method and application
  • Depolymerization glycosaminoglycan extracted from sea cucumber composition and its preparation method and application

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preparation example Construction

[0041] In a preferred embodiment of the preparation of the depolymerized sea cucumber glycosaminoglycan composition of the present invention, in order to obtain a high-purity depolymerized black sea cucumber glycosaminoglycan composition suitable for intravenous injection, the dried black sea cucumber body wall can be twisted successively crushed, centrifuged after enzymatic hydrolysis, the supernatant was precipitated with ethanol, and dried to obtain the precipitate ①; after the precipitate ① was dissolved in water, calcium chloride was added, centrifuged, the supernatant was precipitated with ethanol, and dried to obtain the precipitate ② Dissolve the precipitate ② in water, add potassium acetate, take the supernatant, add ethanol to precipitate, and dry to obtain the crude sea cucumber polysaccharide; pass the crude sea cucumber polysaccharide through a DEAE-cellulose column, use ethanol to precipitate, add hydrogen peroxide to decolorize the heat source , the sea cucumber ...

Embodiment 1

[0047] 1.1 Extraction of sea cucumber polysaccharides

[0048] Take 1000g of dried black sea cucumber (Holothuria atra) body wall, add water to soak, wash and drain, mince, add water to 8kg, adjust the pH value to 8.0 with sodium hydroxide, add alkaline protease (Wuxi Enzyme Preparation Factory) 20ml , at 60° C. for enzymolysis, after the enzymolysis, 2 g of trypsin (Sichuan Deyang Enzyme Preparation Factory) was added for a second enzymolysis, the enzymolysis product was centrifuged at room temperature, and the supernatant was taken. The pH value was adjusted to 2.5 with hydrochloric acid, and the acidic protein precipitate was removed by centrifugation. Add 0.8 times the volume of ethanol, centrifuge at 4°C, and obtain the supernatant to obtain a precipitate. Add 10 times the weight of the precipitate to dissolve it in water, heat to 90°C, add calcium chloride to make the concentration of calcium chloride 2%, keep it warm for 20 minutes, cool, centrifuge, collect the supern...

Embodiment 2

[0069] Example 2 Pharmacodynamic test of depolymerized sea cucumber glycosaminoglycan

[0070] 1. Experimental materials

[0071] 1.1 Animals: SD strain white rats, ♂ sex, weight 250-320g, quality certificate: Shanyidongzhengzi No. 08-005; New Zealand strain white rabbits, ♀♂ half and half, weight 1.8-2.2kg, quality certificate: Shaanxi Medical Dongzheng No. 08-18.

[0072] 1.2 Drug: depolymerized sea cucumber glycosaminoglycan for injection

[0073] Specifications: 10mg / bottle (based on the active ingredient - depolymerized sea cucumber glycosaminoglycan)

[0074] Positive control drug: Xuesaitong injection (specification: 2ml / 100mg / bottle)

[0075] (1) Effects of Depolymerized Sea Cucumber Glycosaminoglycans for Injection on Rats with Focal Cerebral Ischemia

[0076] (1) Experimental method

[0077] Take the SD strain, half male and half female, with enough rats weighing 280-320g, start weighing after 7 days of adaptation to the laboratory, number them, and randomly div...

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Abstract

The invention relates to a de-polymerizing sea cucumber glycosamino-glycan compound, comprising 90-99. 89% of de-polymerizing sea cucumber glycosamino-glycan with weight average molecular weight being between 8000Da to 120000Da. The preparation method comprises following steps: (1) extracting sea cucumber polysaccharides from sea cucumber; (2) degrading extracted polysaccharides; (3) collecting de-polymerizing sea cucumber glycosamino-glycan with weight average molecular weight being between 8000Da to 120000Da. The compound can prevent and treat cerebrovascular disease and cardiovascular disease, especially blood coagulation disease and thrombus disease. The raw material is sea creature, so the source is abundant and cost is low.

Description

technical field [0001] The present invention belongs to the technical field of medicine, specifically, the present invention relates to a depolymerized sea cucumber glycosaminoglycan composition with a molecular weight of 8000Da to 12000Da and a preparation method thereof, as well as a medicine containing the polysea cucumber glycosaminoglycan composition and its preparation method. Application in the prevention and treatment of cardiovascular and cerebrovascular diseases. Background technique [0002] With the increase of population aging, cardiovascular and cerebrovascular diseases have become the biggest threat to human health. According to reports, cardiovascular and cerebrovascular diseases are one of the three major causes of death for the elderly. At present, the deaths due to cardiovascular and cerebrovascular diseases account for 34% of the total death toll, and about 3 / 4 of the surviving patients have varying degrees of labor ability and self-care loss of ability....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/726A61K9/08A61K9/20A61K9/19A61P7/02A61P9/10
Inventor 张登科
Owner XIAN XINTONG PHARM RES CO LTD
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