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Sheep-derived low-molecular-weight heparin and its preparation method and application

A technology of sheep heparin and sheep paparin sodium, applied in the field of medicine and biology

Active Publication Date: 2020-03-24
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-derived low-molecular-weight heparin and its preparation method and application
  • Sheep-derived low-molecular-weight heparin and its preparation method and application
  • Sheep-derived low-molecular-weight heparin and its preparation method and application

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 low-molecular-weight heparin derived from sheep, including sheep dateparin sodium, sheep nadroparin calcium, sheep tinzaparin sodium, sheep betaparin sodium and sheep bemiparin sodium. The low molecular weight heparins of sheep origin above all have common provenance characteristics. In terms of chemical structure, the content of the main disaccharide ΔUA2S‑GlcNS6S (ΔIS) is between 66%‑74%. Sheep low-molecular-weight heparin and pig-derived low-molecular-weight heparin have similar physical and chemical properties and biological activities, which expands the source of low-molecular-weight heparin drugs. The preparation method of sheep low-molecular-weight heparin introduced by the invention is simple and easy, and the process is stable. The obtained product is refined and can fully meet the requirements of various pharmacopoeias for the current low-molecular-weight heparin. Sheep low-molecular-weight heparin also has halal properties that porcine-derived low-molecular-weight heparin does not have. It has a huge market in the majority of Muslim populations, countries and regions, and can be used in anticoagulant, antithrombotic, anti-inflammatory, anticancer and halal 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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Patent Type & Authority Patents(China)
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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