Method for connecting hyaluronic acid and sepharose

A technology of agarose gel and hyaluronic acid, which is applied in the field of organic chemical synthesis, can solve the problems of complex reaction and poor effect, and achieves the effects of simple steps, low cost and mild reaction conditions.

Inactive Publication Date: 2009-12-02
天津市协和医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hyaluronic acid-agarose gel required by the latter needs to connect hyaluronic acid to the agarose gel, the reaction is more complicated, and the general method is not effective

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1. Preparation of hyaluronic acid-adipic acid dihydrazide derivatives: weigh 100 mg of hyaluronic acid, dissolve in 50 ml of double distilled water, add 1 g of adipic acid dihydrazide, stir, and adjust with 0.1M HCl When the pH value was 5.0, 0.5 g of EDC was added, and the pH value was adjusted with 0.1 M HCl to maintain it at 5. After reacting for 2 hours, the pH was adjusted to 7.0 with 1M NaOH. The reacted product was dialyzed with double distilled water for 4 times, each time 1 liter of double distilled water was dialyzed for 18 hours, and then freeze-dried after dialysis.

[0015] 2. Preparation of agarose gel Sepharose 4B containing aldehyde groups: measure 50 ml of agarose gel Sepharose 4B, wash with distilled water, add 10 ml of newly prepared 0.2M NaIO 4 , stirred at room temperature for 5 hours, and the gel after the reaction was washed with distilled water.

[0016] 3. Preparation of derivatives connected between hyaluronic acid and Sepharose 4B: Weigh 20 ...

Embodiment 2

[0018] 1. Preparation of hyaluronic acid-adipic acid dihydrazide derivatives: Weigh 1 g of hyaluronic acid, dissolve it in 500 ml of double distilled water, add 10 g of adipic acid dihydrazide, stir, and adjust the pH with 0.1M HCl When the pH value was 5.0, 5 g of EDC was added, and the pH value was adjusted with 0.1 M HCl to maintain it at 5. After reacting for 2 hours, the pH was adjusted to 7.0 with 1M NaOH. The reacted product was dialyzed with double distilled water for 4 times, each time 1 liter of double distilled water was dialyzed for 18 hours, and then freeze-dried after dialysis.

[0019] 2. Preparation of agarose gel Sepharose 4B containing aldehyde groups: measure 500 ml of agarose gel Sepharose 4B, wash it with distilled water, add 100 ml of newly prepared 0.2M NaIO 4 , stirred at room temperature for 5 hours, and the gel after the reaction was washed with distilled water.

[0020] 3. Preparation of hyaluronic acid and Sepharose 4B linker: Weigh 200 mg of free...

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PUM

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Abstract

The invention belongs to the field of organic chemical synthesis, and relates to a method for connecting hyaluronic acid and sepharose. The method comprises the following steps: synthesizing hyaluronic acid-adipic acid dihydrazide derivatives by using the hyaluronic acid and adipic acid dihydrazide, activating the sepharose 4B by sodium periodate to obtain sepharose containing aldehyde group, and reducing the hyaluronic acid-adipic acid dihydrazide and the sepharose 4B containing the aldehyde group by sodium borohydride to obtain a connection substance of the hyaluronic acid and the sepharose 4B. The method has the advantages of mild reaction condition, simple steps and low cost.

Description

technical field [0001] The invention relates to a method for linking hyaluronic acid and Sepharose 4B, belonging to the field of organic chemical synthesis. Background technique [0002] Hyaluronic acid is widely found in connective tissues of humans and animals. With the continuous emergence of various determination methods, the determination of hyaluronic acid content has reached a precise level, which clearly reflects that the hyaluronic acid in the samples of patients with rheumatoid arthritis, liver disease, certain tumors and scleroderma is significantly increased, especially hyaluronic acid. The determination of hyaluronic acid is superior to all conventional liver function tests in the diagnosis of liver cirrhosis. The hyaluronic acid in serum of patients with liver cirrhosis is 10-100 times higher than that of normal people. It has been reported in China that in chronic liver disease and cirrhosis, the increase of hyaluronic acid is greater than that of type III pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G81/00C08B37/08C08B37/12
Inventor 董全文
Owner 天津市协和医药科技集团有限公司
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