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Hyaluronic acid oligosaccharide fractions and drugs containing the same

A technology of hyaluronic acid and hyaluronic acid, applied in the field of hyaluronic acid oligosaccharide and its new fractions, can solve the problem of no explanation or hint and so on

Inactive Publication Date: 2008-07-30
SEIKAGAKU KOGYO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this publication, it is described that the average molecular weight of HA is preferably 100,000 or more, but there is no description or hint about HA oligosaccharides and their excellent effects.

Method used

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  • Hyaluronic acid oligosaccharide fractions and drugs containing the same
  • Hyaluronic acid oligosaccharide fractions and drugs containing the same
  • Hyaluronic acid oligosaccharide fractions and drugs containing the same

Examples

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Effect test

Embodiment 1

[0213] Preparation and physicochemical properties of oligosaccharides of the present invention and fractions of the present invention

[0214] 1. The preparation of oligosaccharide of the present invention and fraction of the present invention

[0215] The oligosaccharide of the present invention and the fraction of the present invention were prepared using the sodium salt of HA isolated and purified from cockscomb as a raw material by the following method. In addition, the sodium salt of HA as a raw material showed a single band by electrophoresis using a cellulose acetate membrane (electrophoresis buffer: pyridine-formic acid buffer, current: 15 mA, electrophoresis time: 30 minutes), and HA was not detected Other mucopolysaccharides (chondroitin, 4-chondroitin sulfate, 6-chondroitin sulfate, chondroitin sulfate E, chondroitin sulfate D, heparin, heparan sulfate, dermatan sulfate).

preparation example 1

[0216] (Preparation Example 1) Decomposition by Hyaluronidase

[0217] Dissolve 25 g of sodium salt of HA in 1.IL of 0.1 M phosphate buffer (pH 5.3) containing 0.15 M NaCl. To the obtained solution, 200 mg of hyaluronidase (5.342 units / mg; manufactured by Seikagaku Kogyo Co., Ltd.) derived from bovine testis was added and reacted at 37° C. for 9 hours.

[0218] After the reaction, the supernatant was collected by centrifugation at 10,000 rpm for 30 minutes. The recovered supernatant was loaded onto a Dowex 1×2 (100-200 mesh) (manufactured by Dow chemical) ion exchange column (φ1. 5×123cm), eluted by a linear concentration gradient of NaCl (0.01M~0.50M). Fractions containing HA oligosaccharides were screened by detecting uronic acid in the obtained fractions by the carbazole method. Appropriate fractions were pooled and concentrated, desalted using Sephadex G-10 (manufactured by Pharmacia; φ3×124), and then lyophilized.

[0219] The lyophilized weights of the obtained fract...

preparation example 2

[0221] (Preparation Example 2) Decomposition of ACI by Chondroitinase

[0222] Dissolve 8 g of sodium salt of HA in 500 ml of 0.1 M acetic acid buffer (pH6.0) containing 0.1% bovine serum albumin.

[0223] To the obtained solution, 32 units of chondroitinase AC I (manufactured by Seikagaku Kogyo Co., Ltd.) was added and reacted at 37° C. for 6 hours.

[0224] After the reaction, the supernatant was collected by centrifugation at 10,000 rpm for 30 minutes. The recovered supernatant was loaded onto a Dowex 1×2 (100-200 mesh) (manufactured by Dow chemical) ion-exchange column (φ4.5×123 cm), washed by a linear concentration gradient (0.01M-0.50M) of NaCl take off. Fractions containing HA oligosaccharides were screened by detecting uronic acid in the obtained fractions by the carbazole method. Appropriate fractions were pooled, concentrated, desalted with Sephadex G-10 (manufactured by Pharmacia; φ3×124), and then lyophilized.

[0225] The lyophilized weights of the obtained fr...

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Abstract

The present invention provides hyaluronic acid oligosaccharides having a size selected from 4 saccharides to 60 saccharides; fractions containing the hyaluronic acid oligosaccharides and having specific physicochemical properties; and medicines containing these substances. The hyaluronic acid oligosaccharides of the present invention are used as cell death inhibitors, cell disorder inhibitors, and cell and tissue protection agents (such as organ preservation agents, ulcer treatment agents, liver disorder treatment agents, IL-10 production promoters, or IL-8 production inhibitor) is an active ingredient that exerts excellent pharmacological effects and has high safety, so it is extremely useful.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 01812432.1 with the filing date of July 6, 2001. technical field [0002] The present invention relates to hyaluronic acid oligosaccharides (hereinafter referred to as "HA oligosaccharides") and novel fractions thereof. [0003] And the present invention also relates to medicines (especially for heat shock protein expression enhancers, cell death inhibitors, cell barrier inhibitors, or cell and tissue protection agents (especially for organ preservation) using this HA oligosaccharide as an active ingredient. agent, ulcer treatment agent, and liver disorder treatment agent, IL-10 production accelerator, or IL-8 production inhibitor). Background technique [0004] As for hyaluronic acid (hereinafter referred to as "HA"), it is known that its activity or function varies with its molecular size. For example, HA with a molecular weight of 1.2 million can inactivate NF-κb and inhibit angiogenesi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/728C07H7/033C08B37/08A61K31/702A01N1/02A61P43/00A61P1/04A61P1/16
Inventor 浅利晃栗原仁柴田知己宫崎由佳山之口裕子多和田明政孝浩松崎祐二
Owner SEIKAGAKU KOGYO CO LTD
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