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Sheep-sourced low molecular heparins as well as preparation method and application thereof

A technology of low molecular weight heparin and sheep heparin, which is applied in the field of medicine and biology

Active Publication Date: 2018-03-06
SUZHOU RONGXI BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Currently on the market and clinically, there is no low molecular weight heparin prepared from sheep-derived heparin

Method used

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  • Sheep-sourced low molecular heparins as well as preparation method and application thereof
  • Sheep-sourced low molecular heparins as well as preparation method and application thereof
  • Sheep-sourced low molecular heparins as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Preparation and pretreatment of raw material sheep heparin sodium

[0108] Crude sheep heparin sodium (manufacturer: Shandong Shenlian Biotechnology Co., Ltd., batch number: 20150326), the sample was sent for RT-PCR to detect the DNA content of pigs and sheep. The results showed that the DNA content of pigs was less than 0.1 picogram per microliter, and the DNA content of sheep Greater than 1×10 5 picogram per microliter, and according to the source of the original manufacturer's initial intestinal mucosa, it is determined to be crude sheep heparin sodium, and basically does not contain porcine heparin. In addition, the anticoagulant activity of the sample submitted to the whole sheep plasma method was 67.0 units per mg.

[0109] The purification of crude sheep heparin sodium adopts the well-known process in this industry, that is, it is dissolved in water, followed by salt and alkali hydrolysis, and then hydrolyzed with protease. Sheep heparin, the raw material for p...

Embodiment 2

[0112] Preparation of sheep daparin sodium

[0113] Weigh 50.0 g of the pretreated sheep heparin in Example 1, add 500 ml of purified water, and stir to dissolve the heparin. Control the room temperature in a water bath, and continue to stir and keep warm for more than 30 minutes. Adjust the pH of the heparin solution to 3.1 with hydrochloric acid, add 1.43 g of sodium nitrite, and continue to insulate and stir. Then adjust the pH to neutral, add 4.0 g of sodium borohydride, and continue stirring. Then adjust the pH to neutral with dilute hydrochloric acid, add 51.5 g of sodium chloride, and continue stirring at room temperature for more than 10 minutes. Slowly add 1250 ml of methanol and stir for 15 minutes to precipitate the crude dalteparin obtained in the reaction. The reaction solution was transferred to a centrifuge bottle, and the crude dalteparin precipitate was collected by centrifugation. The precipitate was vacuum-dried and weighed to obtain 43.5 g of crude dalte...

Embodiment 3

[0116] Preparation of Sheep Nadroparin Calcium

[0117] Weigh 50.0 g of the pretreated sheep heparin in Example 1, add 500 ml of purified water, and stir to dissolve the heparin. Control the room temperature in a water bath, and continue to stir and keep warm for more than 30 minutes. Adjust the pH of the heparin solution to 3.0 with hydrochloric acid, add 2.5 g of sodium nitrite, and continue to insulate and stir. Then adjust the pH to neutral, add 4.5 grams of sodium borohydride, and continue stirring. Then adjust the pH to neutral with dilute hydrochloric acid, add 52.0 g of sodium chloride, and continue stirring at room temperature for more than 10 minutes. Slowly add 1250 ml of methanol and stir for 15 minutes to precipitate the crude nadrixarin obtained in the reaction. Transfer the reaction solution to a centrifuge bottle, collect the crude nadrixarin precipitate by centrifugation, and weigh the precipitate after vacuum drying to obtain 39.5 g of the crude nadrixarin...

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Abstract

The invention discloses sheep-sourced low molecular heparins which comprise ovine dalteparin sodium, ovine nadroparin calcium, ovine tinzaparin sodium, ovine parnaparin sodium and ovine bemiparin sodium. All the sheep-sourced low molecular heparins have common provenance characteristics; from chemical structures, the content of main disaccharide deltaUA2S-GlcNS6S(deltaIS) is 66 to 74%. The physical and chemical properties of the sheep-sourced low molecular heparins are similar to the physical and chemical properties of pig-sourced low molecular heparins, and the biological activities are close, so that sources of low molecular heparin type drugs are expanded. A preparation method of the sheep-sourced low molecular heparins disclosed by the invention is simple, feasible and stable in process; obtained products are refined, and can completely meet requirements of each pharmacopeia on conventional low molecular heparins. The sheep-sourced low molecular heparins also have the muslim whichis absent in the pig-sourced low molecular heparins, so that the sheep-sourced low molecular heparins have a giant market in general muslim people, countries and districts, and can be applied to anticoagulation, antithrombotic, anti-inflammatory, anti-cancer and muslim drugs.

Description

technical field [0001] The invention relates to low-molecular-weight heparin derived from goats, including goat-daparin sodium, goat-nadroparin calcium, goat-tingzaparin sodium, goat-paparin sodium and goat-bemiheparin sodium, which are halal and belong to the field of medical and biotechnology. Background technique [0002] Low Molecular Weight Heparin (LWMH) is currently the most widely used and most important anticoagulant and antithrombotic drug in clinical practice. Some low molecular weight heparins are also widely used in anti-inflammatory and anti-cancer clinical treatments. Low-molecular-weight heparin is prepared by depolymerization of natural macromolecular-weight heparin. At present, the source of medical low-molecular-weight heparin is almost all porcine intestinal mucosal heparin. [0003] There are certain differences in the chemical structure of heparin derived from different animals or organs. The molecular structure of heparin consists of glucosamine and o...

Claims

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Application Information

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IPC IPC(8): C08B37/10A61K31/727A61P7/02
CPCA61K31/727C08B37/0075
Inventor 金永生靳彩娟姚亦明
Owner SUZHOU RONGXI BIOTECH CO LTD
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