Purification method for reducing protein content in sodium hyaluronate and derivatives thereof

A protein content, sodium hyaluronate technology, applied in the field of pharmaceutical production, can solve the problems of low protein content sodium hyaluronate and its derivatives, unable to meet the sodium hyaluronate quality standards, waste of time and energy consumption, etc., and the process is easy to operate. , Reduce the risk of allergic reactions, low cost effect

Active Publication Date: 2021-05-28
HEFEI BOSIKC PHARMTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] There are great drawbacks in these technological methods, because the technological time of sodium hyaluronate redissolution, complexation and dissociation is too long, generally 8~10 hours, cause the waste of time and energy consumption, multiple alcohol precipitation processes have increased solvent Dosage, at the same time, because the processing time is too long, it may cause the degradation of the sodium hyaluronate macromolecule, and the molecular weight will decrease. A large amount of protein remains, unable to meet the quality standard of sodium hyaluronate, thus affecting the performance of the product
[0016] There are many disadvantages in the purification methods in the prior art. Therefore, there is an urgent need for a purification method that reduces the protein content in sodium hyaluronate and its derivatives, so as to achieve sodium hyaluronate and its derivatives with lower protein content.

Method used

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  • Purification method for reducing protein content in sodium hyaluronate and derivatives thereof
  • Purification method for reducing protein content in sodium hyaluronate and derivatives thereof
  • Purification method for reducing protein content in sodium hyaluronate and derivatives thereof

Examples

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preparation example Construction

[0036]The preparation method of directly extracting sodium hyaluronate from cockscomb refers to the method of patent CN 104610466A. The brief steps are as follows: finely wash the cockscombs after they are thick and fine, mince the cockscombs and perform enzymatic hydrolysis to obtain the enzymatic hydrolysis solution. The protein content of sodium hyaluronate obtained by direct alcohol precipitation of cockscomb enzymatic hydrolysis solution is 1.2% by Folin's phenol reagent method, which is too high and does not meet the quality standard. The examples are experiments after treatment with protein complexing agents.

[0037] Sodium hyaluronate prepared by microbial fermentation is a commercially available product, and the protein content determined by Folin's phenol reagent method is 0.08%.

Embodiment 1

[0038] Embodiment 1: the refining of cross-linked sodium hyaluronate

[0039] Take 1L of cockscomb extract, add 0.2g of inositol hexaphosphate (50% aqueous solution) under stirring, and filter through a 0.22 μm microporous membrane after stirring for 24 hours, add 2L of ethanol to the filtrate, after adding, a white solid appears after stirring Precipitation, suction filtration, dehydration three times, and drying to obtain cross-linked sodium hyaluronate with a protein content of 0.011%.

Embodiment 2

[0040] Embodiment 2: Refining of cross-linked sodium hyaluronate

[0041] Take 1L of cockscomb extract, add 1 g of CPPM (CAS No.2271351-38-1, 1,2,3,4,5-Cyclohexanepentol, 6-[(phosphonooxy)methyl]) under stirring, and pass through 0.22 Filter through a μm microporous membrane, add 2L of ethanol to the filtrate, after the addition is complete, a white solid precipitates out after stirring, filter with suction, dehydrate three times and then dry to obtain cross-linked sodium hyaluronate with a protein content of 0.013%.

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Abstract

The invention discloses a purification method for reducing the protein content in sodium hyaluronate and derivatives thereof. The specific preparation process comprises the following process steps of adding a specific type of protein complexing agent into a sodium hyaluronate and derivative crude product solution, filtering to remove insoluble substances, and carrying out alcohol precipitation to obtain pure products of sodium hyaluronate and derivatives thereof. Under the action of the protein complexing agent, the method has a very good effect of removing proteins in sodium hyaluronate and derivative products thereof, and the content of the proteins in the sodium hyaluronate and the derivatives thereof can be reduced to 0.015% or below, so that the risk of anaphylactic reaction of sodium hyaluronate and derivative preparations thereof caused by heterologous proteins is remarkably reduced; the clinical application value is extremely high; the method disclosed by the invention is simple in technological process, low in cost, easy to operate, good in economical efficiency and more suitable for industrial production; the sodium hyaluronate and the derivative thereof obtained by the invention are high in purity, good in safety and less in anaphylactic reaction, and bring maximum benefits to patients.

Description

technical field [0001] The invention relates to the technical field of pharmaceutical production, in particular to a purification method for reducing protein content in sodium hyaluronate and its derivatives. Background technique [0002] Sodium hyaluronate, also known as sodium hyaluronate or hyaluronic acid, is an acidic mucopolysaccharide. In 1934, Meyer et al. in the United States first isolated the substance from the vitreous body of the bull's eye. Its disaccharide unit is a polymer formed by repeated alternation of D-glucuronic acid and N-acetyl-D-glucose disaccharide units. Repeat The disaccharide units of glycosides are connected into sugar chains by β-1,4 glycosidic bonds, and their weight-average molecular weight is generally 100,000 to 10 million Da. Sodium hyaluronate exists in the lens of the eyeball, mucus in the joint cavity, skin, arterial wall, umbilical cord and other connective tissues in nature, cockscombs and capsules of some bacteria. [0003] Sodium...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
CPCC08B37/0072C08B37/0003
Inventor 郭辉盛荣
Owner HEFEI BOSIKC PHARMTECH CO LTD
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