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Process for preparing high-purity low-molecular heparin sodium

A low-molecular-weight heparin sodium and high-purity technology is applied in the preparation and purification of high-purity low-molecular-weight heparin sodium and biopharmaceutical raw materials. The effect of recovery

Active Publication Date: 2014-06-25
NANJING XINBAI PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above method is a typical industrial process for preparing and purifying heparin sodium, the production process is complex and the cost is high, and there is no mention of the removal method of the main impurity, the structural analogue of heparin sodium chondroitin polysulfate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Dissolve 50 g of crude heparin sodium in 100 mL of 1.2% NaCl solution by mass percentage, mechanically stir until the heparin sodium is completely dissolved, add 1 mol / L NaOH to the solution until precipitation occurs, let stand for 1 hour, filter to remove insoluble matter, and obtain the filtrate; Adjust the pH to 9.5 with 1mol / L NaOH, add 3mL of 1mol / L H 2 o 2 , 10°C, reacted for 2.5 hours. The temperature of the reaction solution was raised to 30°C, and 3 mL of 1mol / L H was added every 3.5 hours thereafter. 2 o 2 , maintain pH 6.8, and react for 21 hours after feeding. Add 5g of active carbon to the reaction solution, magnetically stir, filter to remove insoluble matter, and obtain the filtrate; the Sephadex G-25 column is balanced with a mass percent concentration of 4% NaCl solution, the sample layer is 15% of the column bed volume, and the column diameter / height The ratio is 1:5, the filtrate is passed through the column, and the mass percent concentration is...

Embodiment 2

[0025] Dissolve 50 g of crude heparin sodium in 120 mL of 1.8% NaCl solution by mass percentage, mechanically stir until the heparin sodium is completely dissolved, add 1.5 mol / L NaOH to the solution until precipitation occurs, let stand for 2 hours, filter to remove insoluble matter, and obtain the filtrate ; Adjust the pH to 10.0 with 1.5mol / L NaOH, add 5mL of 3mol / L H 2 o 2 , 20°C, reacted for 3.0 hours. The temperature of the reaction solution was raised to 40°C, and 5 mL of 2mol / L H was added every 2.5 hours thereafter. 2 o 2 , maintain pH 7.0, and react for a total of 18 hours after feeding. Add 5g bentonite to the reaction solution, stir magnetically, remove insoluble matter by filtration, and obtain the filtrate; balance the SephadexG-25 column with a mass percent concentration of 5%NaCl solution, the sample layer is 20% of the column bed volume, and the column diameter / height is 1 : 6, pass the filtrate through the column, elute with a mass percentage concentratio...

Embodiment 3

[0027] Dissolve 50 g of crude heparin sodium in 150 mL of 1.6% NaCl solution by mass percentage, stir mechanically until the heparin sodium is completely dissolved, add 1.2 mol / L KOH to the solution until precipitation occurs, let stand for 1.5 hours, filter to remove insoluble matter, and obtain the filtrate ; Adjust the pH to 10.5 with 1.2mol / L KOH, add 7.5mL of 5mol / L H 2 o 2, 30 ℃, reaction 3.5 hours. The reaction solution was heated to 50°C, after which 7.5mL of 3mol / L H was added every 3 hours 2 O 2 , maintained at pH 7.5, and reacted for a total of 15h after feeding. Add 4.5g bentonite to the reaction solution, stir magnetically, remove insolubles by filtration to obtain a filtrate; equilibrate Sephadex G-25 column with 6% NaCl solution by mass, the sample layer is 25% column bed volume, column diameter / height 1:7, pass the filtrate through the column, elute with a mass percentage concentration of 6% NaCl, the flow rate is one column bed volume / 1.8 hours, collect th...

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Abstract

The invention discloses a process for preparing high-purity low-molecular heparin sodium and belongs to the field of bioengineering. The process mainly comprises the following steps of: salting-out and impurity removal, supplementary material warming and oxidization, gel elution, ion exchange, ethanol precipitation, vacuum drying and the like. The problems that the existing heparin sodium structure analog is hard to remove, the purity can not achieve the requirements for medicinal use, the preparation process is complicated, and the like are solved. The process has the beneficial effects that by adopting a supplementary material warming and oxidization method, the preparation and the decolorization of the low-molecular heparin sodium are realized, and the heparin sodium structure analogue is effectively removed by using weakly-basic anion exchange resin; and the process has the advantages of shortened production period, saved cost, simple process, easiness in operation and is suitable for industrial production.

Description

technical field [0001] The invention relates to a method for preparing and purifying biomedicine raw materials, which belongs to the field of bioengineering, in particular to a process for preparing high-purity low-molecular-weight heparin sodium. Background technique [0002] Heparin sodium is an anion-rich mucopolysaccharide sulfate, which is produced by mast cells and basophils in animals. It is originally extracted from the liver, so it is named heparin. Clinically used heparin is sodium (lithium) salt extracted from animal intestines, lung mucosa, and blood vessel walls. Heparin sodium (lithium) has various biological functions such as anticoagulation, anti-inflammation, and anti-cancer, but long-term use There will be side effects such as bleeding and fractures. In the early 1980s, low molecular weight heparin sodium (LMWH, <8000Da) was found to have stronger activity and less side effects, and became the main clinical anticoagulant drug. [0003] At present, more...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/10
Inventor 胡永红沈飞于辉李栋芸冯旌李佼佼杨文革
Owner NANJING XINBAI PHARMA
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