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Method for connecting hyaluronic acid and sepharose

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

Inactive Publication Date: 2010-12-08
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 derivative: Weigh 100 mg of hyaluronic acid, dissolve it in 50 ml of double-distilled water, add 1 g of adipic acid dihydrazide, stir, and adjust with 0.1 M HCl At pH 5.0, 0.5 g of EDC was added and the pH was adjusted to maintain at 5 with 0.1 M HCl. After 2 hours of reaction, the pH was adjusted to 7.0 with 1M NaOH. The reacted product was dialyzed against double distilled water for 4 times, each time with 1 liter of double distilled water for 18 hours, and freeze-dried after dialysis.

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

[0016] 3. Preparation of hyaluronic acid and Sepharose 4B linked derivatives: Weigh 20 mg of freeze-dried hyaluronic acid-adipic ...

Embodiment 2

[0018] 1. Preparation of hyaluronic acid-adipic acid dihydrazide derivative: 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 pH with 0.1 M HCl 5.0, 5 g of EDC was added, and the pH was adjusted to 5 with 0.1 M HCl. After 2 hours of reaction, the pH was adjusted to 7.0 with 1M NaOH. The reacted product was dialyzed against double distilled water for 4 times, each time with 1 liter of double distilled water for 18 hours, and freeze-dried after dialysis.

[0019] 2. Preparation of Sepharose 4B containing aldehyde groups: Measure 500 ml of Sepharose 4B agarose, wash with distilled water, add 100 ml of newly prepared 0.2M NaIO 4 , the reaction was 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 conjugate: Weigh 200 mg of freeze-dried hyaluronic acid-adipic acid dihydrazide derivati...

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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, andreducing 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 sepharose4B. 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 connecting hyaluronic acid and agarose gel Sepharose 4B, and belongs to the field of organic chemical synthesis. Background technique [0002] Hyaluronic acid is widely found in the connective tissue of humans and animals. The continuous emergence of various assay methods has made the determination of hyaluronic acid content to an accurate 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 the transparent The determination of hyaluronic acid is superior to all conventional liver function tests in the diagnosis of liver cirrhosis. The serum hyaluronic acid 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 II...

Claims

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Application Information

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Patent Type & Authority Patents(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