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Preparation method of endotoxin absorbent for blood perfusion

A hemoperfusion and adsorbent technology, applied in the field of preparation of endotoxin adsorbent, can solve problems such as ineffective effects, achieve the effects of improving biocompatibility, increasing safety, and overcoming toxicity and instability

Inactive Publication Date: 2006-11-22
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestically, there are reports of animal experiments using activated carbon to adsorb endotoxin, but the effect is not obvious [Wang Jinda. China Critical Emergency Medicine, 10: 578 (1998)]; Nankai University uses agarose as a carrier, after epoxidation, amination, A series of adsorbents for removing endotoxin in plasma were prepared by bonding affinity ligands after dialdehyde cross-linking [Yuan Zhi et al., Chinese Invention Patent, publication number: CN1493368A, 2004]

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1) Preparation of epoxy matrix

[0025] Use a graduated cylinder to measure 50 mL of Sepharose CL-4B agarose gel, pour it into a sand core funnel, rinse it with water, and drain it. Transfer to a Erlenmeyer flask, add 50mL of 1mol / L NaOH and 25mL of epichlorohydrin, seal it, and react in a shaker at 40°C and 200r / min for 2 hours. After the reaction is completed, fully rinse with water, filter and dry for later use;

[0026] 2) Preparation of Amino Matrix

[0027] Transfer the above-mentioned epoxy base into the Erlenmeyer flask, add 0.1mol / L NaHCO of 13g / L hexamethylenediamine 3 Buffer 250mL. Shaker at 50-55°C, 150r / min, react for 2 hours. After the reaction is completed, pour it into a sand core funnel for suction filtration, fully rinse with water, and dry it for later use;

[0028] 3) Immobilization of affinity ligands

[0029] Take a certain volume of dioxane in a round-bottomed flask, add an appropriate amount of metal sodium, heat and reflux for 6-12 hours, ...

Embodiment 2

[0032] Embodiment 2 Adsorbent is used for the removal of endotoxin in aqueous solution

[0033] Take 50mg of polymyxin B-bound adsorbent, add 1mL 16Eu / mL endotoxin solution, shake at 25°C for 2 hours, centrifuge to get the supernatant, and use the azochromogenic method to measure the concentration of endotoxin. Experimental results show that the removal rate of endotoxin is greater than 90%.

Embodiment 3

[0034] Example 3 Comparison of the bonding amount of two endotoxin adsorbents polymyxin B and the removal results of endotoxin in aqueous solution

[0035] Adsorbent 1: Sepharose CL-4B agarose gel is activated by epichlorohydrin and directly bonded with polymyxin B to obtain an endotoxin adsorbent.

[0036] Adsorbent 2: Sepharose CL-4B agarose gel is activated by epichlorohydrin, hexamethylenediamine, and 1,1'-carbonyldiimidazole, and then bonded with polymyxin B to obtain an endotoxin adsorbent.

[0037] Polymyxin B binding amount (mg / g)

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PUM

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Abstract

The invention relates to an endotoxin adsorbent, which in detail relates to a method of preparing endotoxin adsorbent for blood perfusion. The method employs spherical agarose gel with good biocompatibility as base material, employs epichlorohydrin, hexamethylene diamine, 1, 1'-carbonyldiimidazole as activating agent, and bonds polymyxin B for endotoxin removal. The invention is characterized in that it overcomes the toxicity of cyanogen bromide and instability of aglycone, improves the safty and reliability of operation, reducesnon-specific adsorption through bonding polymyxin B with 1, 1'-carbonyldiimidazole, and improves biocompatibility of adsorbent and suits for treating endotoxemia. The invention is also characterized by simple experimental procedure and high clearance for endotoxin.

Description

technical field [0001] The invention relates to an endotoxin adsorbent, in particular to a preparation method of an endotoxin adsorbent for hemoperfusion. Background technique [0002] Endotoxin is a kind of lipopolysaccharide produced by cleavage of the cell wall when Gram-negative bacilli grow or die. A very small amount (nanogram level) of endotoxin entering the human body will cause high fever, diarrhea, vasodilation, and even fainting or die. Endotoxemia is one of the most common clinical fatal diseases, the mortality rate can reach 40%-90%, and there is no effective treatment method in clinical practice. How to timely and effectively remove the endotoxin in the patient's body is the key to the treatment of endotoxemia. In recent years, a variety of molecular biological agents to antagonize endotoxin have been developed abroad, but the production process is complicated, the drug sources are few, and the cost is high, and so far none of them has passed the phase III cl...

Claims

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Application Information

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IPC IPC(8): A61M1/34A61L33/04A61M1/36
Inventor 邹汉法唐守万孔亮
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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