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A purification method for reducing protein content in sodium hyaluronate and its derivatives

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, failure to meet the quality standards of sodium hyaluronate, waste of time and energy consumption, etc., to achieve easy operation of the process , reduce the risk of allergic reactions, low cost effect

Active Publication Date: 2022-06-28
HEFEI BOSIKC PHARMTECH CO LTD
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  • 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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  • A purification method for reducing protein content in sodium hyaluronate and its derivatives
  • A purification method for reducing protein content in sodium hyaluronate and its derivatives
  • A purification method for reducing protein content in sodium hyaluronate and its derivatives

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preparation example Construction

[0036]The preparation method of directly extracting sodium hyaluronate from chicken comb is prepared with reference to the method of patent CN 104610466A. The brief steps are as follows: the thickness of the cockscomb is finely washed again, the cockscomb is minced and then enzymatically hydrolyzed, and the enzymatic hydrolysis solution is obtained. The protein content of sodium hyaluronate obtained by direct alcohol precipitation of the cockscomb enzymolysis solution was 1.2% by the Folin phenol reagent method. The protein content was too high and did not meet the quality standard. Examples are experiments after treatment with protein complexing agents.

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

Embodiment 1

[0038] Example 1: Refinement of cross-linked sodium hyaluronate

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

Embodiment 2

[0040] Example 2: Refinement of cross-linked sodium hyaluronate

[0041] Take 1 L of cockscomb extract, add 1 g of CPPM (CAS No.2271351-38-1, 1,2,3,4,5-Cyclohexanepentol,6-[(phosphonooxy)methyl]) under stirring, stir for 0.5 hours and pass through 0.22 Filtration with a μm microporous membrane, adding 2L of ethanol to the filtrate, after the addition was completed, a white solid was precipitated after stirring, suction filtration, dehydration three times, and drying 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 its derivatives. The specific preparation process includes the following process steps: adding a specific type of protein complexing agent to the crude product solution of sodium hyaluronate and its derivatives, filtering The insoluble matter is removed, and pure sodium hyaluronate and its derivatives are obtained by alcohol precipitation. The present invention has a good effect on removing the protein in sodium hyaluronate and its derivative products through the function of protein complexing agent, and can reduce the protein content in sodium hyaluronate and its derivative products to below 0.015%, thus significantly It reduces the risk of allergic reactions caused by heterologous proteins to sodium hyaluronate and its derivative preparations, and has extremely high clinical application value; the method of the invention has simple process, low cost, easy operation, good economy, and is more suitable for industrialization Production; the sodium hyaluronate and its derivatives obtained in the present invention have high purity, good safety, less allergic reactions, 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 the 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. first isolated this substance from the bovine vitreous body. Its disaccharide unit is a polymer formed by repeated alternation of D-glucuronic acid and N-acetyl-D-glucose disaccharide units. The disaccharide units are connected by β-1,4 glycosidic bonds to form sugar chains, and the weight average molecular weight is generally 100,000 to 10 million Da. In nature, sodium hyaluronate mostly exists in the eye lens, joint cavity mucus, skin, arterial wall, umbilical cord and other connective tissues, cockscombs and the capsule of some bacteria. [0003] Sodium hyaluronate is a polymer compound, and its repeating ...

Claims

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Application Information

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