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Process for refining low-molecular dermatan sulfate by enzyme-ultrafiltration method and application

A low-molecular-weight dermatan sulfate technology is applied to organic active ingredients, medical preparations containing active ingredients, blood diseases, etc. It can solve the problems of slow hydrolysis reaction, low bioavailability, excessive hydrolysis, etc. High product purity, high process repeatability, and low ultrafiltration loss

Inactive Publication Date: 2020-04-21
DONGYING TIANDONG PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As for the source of dermatan sulfate, natural dermatan sulfate can be used directly, which is mainly derived from animal tissue extraction. Since it is mixed with other glycosaminoglycans, proteins, nucleic acids and other components in animals, it is difficult to extract and purify
In addition, the relative molecular mass of natural DS is generally between 15,000 and 45,000 Da, which is too large to be absorbed by the human body. If it is directly used in the human body, there are two main pharmacokinetic problems: one is the short half-life, and the other is the subcutaneous and intramuscular When injected internally, its bioavailability is low; and the higher the relative molecular weight of DS, the lower its bioavailability
Dermatan sulfate is hydrolyzed in hydrochloric acid solution, but the amount of hydrochloric acid used is large, the hydrolysis reaction is slow, and the hydrolysis reaction is not easy to control, which may cause excessive hydrolysis or insufficient hydrolysis
The use of hydrogen peroxide to oxidize and degrade dermatan sulfate also has problems such as difficult control of degradation parameters, poor reaction stability and repeatability, resulting in low yield, uneven molecular weight distribution of degradation products, and uncontrollable curative effect.

Method used

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  • Process for refining low-molecular dermatan sulfate by enzyme-ultrafiltration method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0155] Weigh 1Kg of heparin sodium by-product and add 10L of water to dissolve it into a feed liquid with a concentration of about 10%, raise the temperature of the feed liquid to 60°C, and slowly add about 10L of resin to the resin column for resin adsorption; after the adsorption is completed, use 25L concentration of 3.5% sodium chloride solution, adjust the temperature to 60°C for elution, discard the eluent; then use 30L concentration of 8% sodium chloride solution (adjust temperature to 60°C) for elution, Collect this part of the eluate to the 1# tank; after the elution is completed, accurately measure the volume of the collected eluate to be 32L, add 12.8L of ethanol, stir evenly, let it stand for 3 hours, and transfer the supernatant suspension to the 2# tank (the sediment at the bottom of the 1# tank is heparinoid), add 64L of ethanol to the 2# tank again, stir evenly, let it stand for 6 hours, discard the supernatant, collect the sediment at the bottom of the 2# tank,...

Embodiment 2

[0164] Weigh 50Kg of the heparin sodium by-product and add 500L water to dissolve it, and dissolve it into a feed liquid with a concentration of about 10%, raise the temperature of the feed liquid to 60°C, slowly add about 500L resin to the resin column, and perform resin adsorption; after the adsorption is completed, use 1250L concentration of 3.5% sodium chloride solution, adjust the temperature to 60°C for elution, discard the eluent; then use 1500L concentration of 8% sodium chloride solution (adjust temperature to 60°C) for elution, Collect this part of the eluate to the 5# tank; after the elution is completed, accurately measure the volume of the collected eluate to be 1600L, add 640L of ethanol, stir evenly, let stand for 3h, and transfer the supernatant suspension to the 6# tank (The sediment at the bottom of the 5# tank is heparinoid), add 3000L ethanol to the 6# tank again, stir evenly, let it stand for 8 hours, discard the supernatant, collect the sediment at the bot...

Embodiment 3

[0170] Weigh 100Kg of the heparin sodium by-product and add 1000L water to dissolve it, and dissolve it into a feed liquid with a concentration of about 10%, raise the temperature of the feed liquid to 60°C, and slowly add about 1000L of resin to the resin column for resin adsorption; after the adsorption is completed, use 2500L concentration of 3.5% sodium chloride solution, adjust the temperature to 60°C for elution, discard the eluent; then use 3000L concentration of 8% sodium chloride solution (adjust temperature to 60°C) for elution, Collect this part of the eluate to the 5# tank; after the elution is completed, accurately measure the volume of the collected eluate to be 3200L, add 1280L of ethanol, stir evenly, let stand for 3h, and transfer the supernatant suspension to the 6# tank (The sediment at the bottom of the 5# tank is heparinoid), add 6000L ethanol to the 6# tank again, stir evenly, let it stand for 8 hours, discard the supernatant, collect the sediment at the b...

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Abstract

The invention provides a low-molecular dermatan sulfate. The number-average molecular weight of the low-molecular dermatan sulfate is 2,000 to 5,000 Da, and the weight-average molecular weight of thelow-molecular dermatan sulfate is 2500 to 6500 Da. The refined low-molecular dermatan sulfate obtained by the invention has narrower weight-average molecular weight distribution and data molecular weight distribution, is more suitable for human body absorption, and has a wide application prospect in antithrombotic drugs. The invention provides a process for refining the low-molecular dermatan sulfate. The process is a process for refining the low-molecular dermatan sulfate by an enzyme-ultrafiltration method. Particularly, the heparin sodium byproduct rich in dermatan sulfate is used as a rawmaterial, a bi ological enzyme-chondroitin sulfate B enzymecapable of degrading dermatan sulfate is utilized, an ultrafiltration method is combined, the comprehensive refining process is obtained, thereaction is mild, stable and controllable, the process repeatability is high, certain economic benefits and environmental protection effects are achieved, and the comprehensive refining process is suitable for industrial application and promotion.

Description

technical field [0001] The invention belongs to the technical field of dermatan sulfate, and relates to a low-molecular-weight dermatan sulfate and its refining process and application, in particular to a low-molecular-weight dermatan sulfate and the process and application of refining the low-molecular-weight dermatan sulfate by an enzyme-ultrafiltration method. Background technique [0002] Dermatansulfate (DS for short) is the most widely distributed extracellular matrix glycosaminoglycan in animals, and it is the main component of proteoglycan in blood vessel walls. As a glycosaminoglycan, dermatan sulfate is widely distributed in animal tissues. As an important part of connective tissue, DS has a variety of pharmacological effects and physiological functions, and can be used as medicine and health food, mainly for osteoarthritis (oa) and coronary atherosclerotic heart disease (coronary heart disease). The antithrombotic effect is equivalent to or even stronger than tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08C12P19/26A61K31/737A61P7/02
CPCA61K31/737A61P7/02C08B37/0069C12P19/26
Inventor 董磊李玮涛艾自明孙守政
Owner DONGYING TIANDONG PHARM CO LTD
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