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Preparation method for hyaluronic acid, and anti-adhesive composition comprising hyaluronic acid prepared by same preparation method

A technology of hyaluronic acid and manufacturing method, which is applied in the field of manufacturing, can solve the problems of short residence time and achieve the effect of slow decomposition speed and excellent anti-adhesion effect

Active Publication Date: 2016-02-10
IL DONG PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in the prior art in Korea, there is no case of using hyaluronic acid alone without cross-linking or changing the structure; for the use of hyaluronic acid, it is generally believed in the prior art in Korea that Hyaluronic acid is easily decomposed in the body, so the residence time is short, and most people hold a negative view that only hyaluronic acid that has not been cross-linked is used

Method used

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  • Preparation method for hyaluronic acid, and anti-adhesive composition comprising hyaluronic acid prepared by same preparation method
  • Preparation method for hyaluronic acid, and anti-adhesive composition comprising hyaluronic acid prepared by same preparation method
  • Preparation method for hyaluronic acid, and anti-adhesive composition comprising hyaluronic acid prepared by same preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Anti-adhesion effect experiment brought by molecular weight

[0066] SD rats (female, SPF, Oriental Bio Co., Ltd.) were anesthetized by inhalation, and the anesthesia was continued during the operation. The limbs were fixed on the operating table, the hair on the abdomen was removed with a razor, and the abdomen was cut open with scissors after disinfection. The cecum was taken out, and wounds were made with a roving cloth to the extent that the blood seeped onto the gauze, and the condition that the cecum adhered to the abdominal wall where it was located was created, or the condition that the cecum adhered to the inner peritoneal wall was created by the same method. After the sample was processed, it was sutured, and after 2 weeks of feeding, the abdomen was opened to confirm the state of adhesion. The samples were used by dissolving hyaluronic acid having an average molecular weight of 3 million Da, 4 million Da, and 6 million Da in a sodium phosphate buffer solut...

Embodiment 2

[0069] Basic Culture Conditions for Production of High Molecular Hyaluronic Acid

[0070] Quickly thaw 4ml of the culture solution of Streptococcus dysgalactiae ID9103 strain stored in a freezer at -72°C, spread it on 5.2% brain heart infusion solid medium, cultivate it at 37°C for 24 hours, and cut out 1 cm in length and width. 2 Bacterial colonies grown in the area were inoculated into 3% Todd-Hewitt broth sterilized liquid medium (heart infusion 0.31%, neoptone peptone 2%, glucose 0.2%, disodium hydrogen phosphate 0.04%, sodium carbonate 0.25% ; BD Company, USA) in 40ml.

[0071]40 ml of the culture solution cultured with shaking at 37° C. and 150 rpm was used as the primary seed culture solution. In a 6-hour culturing state in the logarithmic growth stage, inoculate the primary inoculum solution into three 3% Todd-Hewitt broth sterilized liquid medium (40 ml, pH 7.8) in an aseptic manner. The culture conditions were aseptic culture at 37° C. and 150 rpm for 20 hours or...

Embodiment 3

[0076] High Molecular Hyaluronic Acid Productivity and Molecular Weight

[0077] Nitrogen sources are known to play an important role in microbial metabolism and also affect the production of hyaluronic acid. Therefore, it was judged that the production of hyaluronic acid of 4 million Da to 6 million Da can be contributed by changing the amino acids containing peptone as the nitrogen source used basically.

[0078] It was confirmed that the production amount and molecular weight of hyaluronic acid differed depending on the peptones, and among various peptones, casein (enzyme hydrolyzate) as a base medium source showed a production amount exceeding 8 g / L and Molecular weight of 4.32 million Da, and showed the best results, so in the tests of the following examples, casein was also used.

[0079] In the case of the group in which arginine was added instead of glutamine as the basic medium, the production amount and molecular weight of hyaluronic acid showed excellent results of...

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Abstract

The present invention relates to a preparation method for hyaluronic acid having a low degradation rate in the body, and an anti-adhesive composition comprising hyaluronic acid prepared by the preparation method. More specifically, the present invention provides a preparation method for hyaluronic acid having a low degradation rate in the body, comprising a step for culturing Streptococcus dysgalactiae strain 9103 (KCTC11818BP) in a medium comprising a carbon source and a nitrogen source, and an anti-adhesive composition comprising hyaluronic acid prepared by the preparation method. The hyaluronic acid prepared by the preparation method of the present invention is a high-molecular hyaluronic acid of 3.5 million Da to 10 million Da, and has an excellent anti-adhesive effect due to slow degradation rate in the body. The anti-adhesive composition of the present invention, comprising hyaluronic acid prepared by the preparation method of the present invention, is capable of improving the problems of conventional anti-adhesive compositions comprising crosslinking agents and compounds, and the problems of conventional hyaluronic acid, by using non-crosslinked hyaluronic acid.

Description

technical field [0001] This application claims the priority of Korean Patent Application No. 10-2013-0029876 (Application No.) filed on March 20, 2013, the entirety of which is hereby incorporated by reference. [0002] The present invention relates to a method for producing hyaluronic acid having a low decomposition rate in vivo and an anti-adhesion composition containing hyaluronic acid produced by the above method. A method for producing hyaluronic acid having a low decomposition rate in vivo by culturing a strain (KCTC11818BP) in a medium containing carbon and nitrogen sources, and an anti-adhesion composition containing hyaluronic acid produced by the method. Background technique [0003] Hyaluronic acid (Hyaluronicacid (HA), Hyaluronan, (C 14 h 20 NNaO 11 )n(n>1000)) are macromolecules present in living organisms and are polysaccharides of glycosaminoglycans. In its structure, D-glucuronic acid and N-acetylglucosamine are repeatedly linked by -1,3 bonds and -1,4...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04C08B37/08A61K31/728
CPCC12P19/26A61L27/20A61L27/58A61L31/042A61L31/148C08L5/08C08B37/0072A61K31/728A61P43/00C12P19/04C12N1/205C12R2001/46
Inventor 姜大中金兑允康在勋
Owner IL DONG PHARMA CO LTD
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