CN112569910B

CN112569910B - Preparation method of adsorbent for removing LDL (low density lipoprotein) by blood extracorporeal circulation and perfusion device

Info

Publication number: CN112569910B
Application number: CN202011254458.1A
Authority: CN
Inventors: 董凡; 谢煜
Original assignee: Jafron Biomedical Co Ltd
Current assignee: Jafron Biomedical Co Ltd
Priority date: 2020-11-11
Filing date: 2020-11-11
Publication date: 2023-07-04
Anticipated expiration: 2040-11-11
Also published as: CN112569910A

Abstract

The invention relates to a preparation method of an adsorbent for removing LDL (low density lipoprotein) by blood in vitro circulation and a perfusion device, wherein the preparation method comprises the following steps: step one: carrying out ring opening reaction on the carrier containing amino and halogenated epoxy compound to obtain intermediate product containing halogenated hydrocarbon group; step two: and (3) carrying out acidolysis reaction on the intermediate product and polyanion to obtain the adsorbent. The preparation method provided by the invention can improve the production efficiency and the safety of products.

Description

Preparation method of adsorbent for removing LDL (low density lipoprotein) by blood extracorporeal circulation and perfusion device

Technical Field

The invention relates to the technical field of extracorporeal blood purification, in particular to an adsorbent for removing low-density lipoprotein, total cholesterol and triglyceride by extracorporeal blood circulation, a preparation method thereof and a perfusion device.

Background

Lipid metabolism disorders are important causative agents of atherosclerosis, which in turn leads to cardiovascular and cerebrovascular diseases, the correlation of which has been confirmed by a number of animal experiments, histopathological and histochemical studies of human atheromatous plaques, and numerous epidemiological surveys.

Active lipid-lowering therapy plays an important role in cardiovascular and cerebrovascular diseases. Most patients with lipid metabolism disorder can well control diseases through active diet control, proper physical activity and proper lipid regulating drugs, but congenital homozygous familial hypercholesterolemia patients need timely and timed lipid-lowering treatment, otherwise coronary heart disease occurs in early years, and the service life is obviously shortened. For patients with severe lipid metabolism disorders and microcirculation disorders combined with partial acute ischemic vascular diseases, which are in need of lipid lowering, and for patients with particularly high blood cholesterol, undesirable blood lipid lowering after conventional treatment or intolerance to drug treatment and serious side effects, this population requires more effective in vitro blood lipid purification treatment, and in such a background, many in vitro lipid lowering therapies have developed.

The treatment of patients with hyperlipidemia by blood purification therapy can rapidly and effectively remove Low Density Lipoprotein (LDL), the therapeutic effect of which has been approved, and the technology of which has been very mature abroad. At present, the domestic blood purification therapy for removing LDL is still in the popularization and development stage.

The current commercial and reported preparation method of LDL adsorbents in the world mainly comprises the steps of immobilizing polyanion, polyamino acid, phosphate groups and the like on carriers such as synthetic resin, porous composite fiber and the like, and compared with the method, the adsorbent prepared by immobilizing the polyanion on the synthetic resin has better comprehensive performance, such as DALI products (polyacrylic acid immobilized on porous polyacrylamide carriers) of Fresenius company in Germany. The synthetic adsorbent disclosed in patent US5476715a of Fresenius, germany, has the process route: mixing the aminated carrier with polyacrylic acid and EEDQ, controlling pH, reacting for 12h at normal temperature, cleaning and purifying. EEDQ is also disclosed as a condensing agent in patent application CN109248668A by Jianfan Biotechnology group Co., ltd. The condensing agent EEDQ is very unstable, the condensing agent EEDQ needs to be stored and transported at the temperature of 2-8 ℃, the temperature is increased to easily generate decomposition reaction to fail, in the reaction process, in order to maintain the activity of the EEDQ, the reaction temperature is set to be room temperature, the reaction temperature is relatively low, the reaction time is prolonged to be more than 12 hours, and the reaction speed is slow. EEDQ and its decomposed products are insoluble in water and most organic solvents, are not easily removed from LDL adsorbents, remain in the product to affect the safety of the product, and byproducts such as quinoline have a pungent smell. Moreover, the reaction typically requires an excess of EEDQ to promote the reaction, and excess condensing agent may remain directly. In addition, in the course of blood purification treatment, the patient's blood must be directly contacted with an LDL adsorbent and returned to the patient directly after removal of LDL, and in order to reduce adverse effects of treatment, it is necessary to introduce as little foreign substances as possible into the blood in addition to a high LDL clearance rate, and therefore it is necessary to reduce various kinds and amounts of residual substances in the LDL adsorbent.

Disclosure of Invention

In view of the shortcomings of the prior art, a first object of the present invention is to provide a method for preparing an adsorbent for removing LDL by extracorporeal circulation of blood, which has simple steps, easy reaction, mild and adjustable reaction conditions, easy reduction of reaction time, and easy removal and separation of residues. A second object of the present invention is to provide a perfusion apparatus comprising the above adsorbent.

To achieve the first object of the present invention, the present invention provides a method for preparing an adsorbent for extracorporeal circulation of blood for removing LDL, comprising the steps of: step one: carrying out ring opening reaction on the carrier containing amino and halogenated epoxy compound to obtain intermediate product containing halogenated hydrocarbon group; step two: and (3) carrying out acidolysis reaction on the intermediate product and polyanion to obtain the adsorbent.

From the above, the invention is characterized in that halogenated hydrocarbon groups are introduced into the carrier through the rapid ring-opening reaction of halogenated epoxy compounds and the carrier containing amino groups, and then the acid hydrolysis reaction is carried out on halogen atoms and polyanions, so that the polyanions are covalently connected to the carrier, and the adsorbent grafted with the polyanions on the carrier is obtained, and the adsorbent can adsorb positively charged low-density lipoprotein under the normal physiological pH environment through the negative charges of the polyanions, and has good adsorption effect on total cholesterol and triglyceride. Compared with the method for realizing grafting by promoting the condensation of carboxyl and amino by a condensing agent such as EEDQ in the prior art, the preparation method has various advantages: the method has the advantages of simple reaction steps, easy reaction, no need of catalyst, high reaction rate, short reaction period, mild reaction conditions, adjustable reaction conditions at normal temperature, for example, the reaction temperature can be increased, thereby being convenient for shortening the reaction time and improving the production efficiency; moreover, the residues are easy to separate and clean, for example, the residues can be cleaned by excessive or water washing, the residues are convenient to collect or recycle, and almost no byproducts are produced or the byproducts have no safety influence in the reaction process, so that the problem of byproduct residues is avoided.

In the second step, the intermediate product is mixed with the aqueous solution of the polyanion and then subjected to acidolysis reaction.

As can be seen from the foregoing, the polyanion used in the present invention undergoes acidolysis reaction with the intermediate product in the form of an aqueous solution of the polyanion. The water is used as a solvent, so that the polyanion can be dispersed to better react with the intermediate product containing halogenated hydrocarbon groups, the method is environment-friendly and pollution-free, and the solvent and the excessive polyanion can be easily removed by adopting methods such as filtration, water washing and the like after the reaction is finished.

The further technical proposal is that the mass ratio of the polyanion to water in the aqueous solution of the polyanion is (1.5-3): 5, preferably 1.5:5.

from the above, the invention further limits the mass ratio of the polyanion to water in the polyanion aqueous solution, and when the mass ratio is adopted, the concentration of the polyanion is proper, so that the carboxyl participating in the reaction on each polyacrylic acid molecular chain is moderate, and excessive consumption of the functional group carboxyl can not be caused, thereby ensuring better adsorption performance.

Further technical proposal is that the pH value of the aqueous solution of the polyanion is 7-8.

As is apparent from the above, the polyacrylic acid solution of the present invention can be adjusted to pH 7 to 8 with an alkaline substance such as sodium hydroxide in advance so that the carboxyl group of the polyacrylic acid is COO ^－ In the form of (2) is readily reacted with the halogenated hydrocarbon groups on the support of the intermediate product.

In the second step, the acidolysis reaction temperature is 35-60 ℃.

Therefore, the reaction process of the invention can be properly heated as required, the temperature rise can not affect the property and performance of materials, and the problems of low reaction temperature and limited reaction rate caused by unstable condensing agent in the prior art are overcome.

Further technical proposal is that the polyanion is at least one selected from polyacrylic acid, carbomer and dextran sulfate.

From the above, the invention further defines the species of polyanion. Preferably, the polyanion is a polyanion disclosed in chinese patent application CN109248668A, the polyacrylic acid is a polyacrylic acid having a molecular weight of 5000 to 450000 or a mixture of at least two polyacrylic acids having different molecular weights, the carbomer is a single-brand carbomer or a mixture of at least two different-brand carbomers, the dextran sulfate is subjected to hydroformylation with periodate before reacting with amino groups and reduction after reacting with amino groups, the polyanion is used to improve the adsorptivity of the adsorbent to LDL and the like, and the adsorbent is reduced to beneficial metal ions such as Ca in blood ²⁺ 、Mg ²⁺ And the like, or to improve the binding firmness of the polyanion and the carrier.

Further technical proposal is that the halogenated epoxy compound is at least one selected from epichlorohydrin, methyl epichlorohydrin and 1, 2-epoxy chlorobutane.

As is apparent from the above, the halogenated epoxy compound of the present invention is preferably of the above-mentioned type, and the raw materials are readily available and low-cost, and can be dissolved in ethanol, so that the residual excess halogenated epoxy compound can be removed by washing with ethanol and water after the reaction.

In the first step, the carrier containing amino, halogenated epoxy compound and water are mixed and then ring-opening reaction is carried out; the volume of water is 2 to 5 times the volume of the halogenated epoxy compound.

From the above, the first step of the present invention can be performed in the presence of an aqueous solvent, which is conducive to dispersing the halogenated epoxy compound by stirring to facilitate the reaction, and the aqueous solvent is environmentally friendly and pollution-free, so that most of the solvent and unreacted halogenated epoxy compound can be removed by filtration or the like after the reaction.

The further technical scheme is that the method further comprises the step a) before the step one: the carrier containing epoxy group and amine are subjected to ring opening reaction to prepare the carrier containing amino group in the first step.

From the above, the invention also provides a preparation step of the carrier containing the amino, wherein the epoxy group can be connected to the carrier, and the amination of the carrier can be realized through the reaction of the epoxy group and the amine compound.

In the step a, the ring-opening reaction is carried out after the carrier containing the epoxy group, the amine and the water are mixed.

From the above, it can be seen that when preparing an amino group-containing carrier from an epoxy group-containing carrier, the reaction can be carried out in the presence of an aqueous solvent, which facilitates the dispersion of the amine to facilitate the reaction, and the aqueous solvent is environmentally friendly and pollution-free, which facilitates the removal of the solvent and unreacted amine by filtration or the like after the reaction.

In the step a, the carrier containing the epoxy group is selected from at least one of macroporous styrene resin containing the epoxy group, macroporous acrylic resin and large Kong Qianwei element spheres.

As described above, the carrier of the present invention may be selected from the conventional resin or cellulose carriers, and the carrier may have an active group such as a carboxyl group or a hydroxyl group, for example, and may be capable of reacting with an epoxidizing agent such as epichlorohydrin or 1, 4-butanediol diglycidyl ether to introduce an epoxy group. The carrier preferably has a macroporous structure, which increases the specific surface area of the adsorbent, and increases the adsorption rate and adsorption capacity.

In the step a, the amine is at least one of ammonia water, ethylenediamine, 1, 2-propylenediamine and 1, 3-propylenediamine.

From the above, the amine used in the preparation of the carrier containing an amino group is preferably a lower amine compound, which has good water solubility, is easily removed by washing after the reaction, and has low raw material cost.

The further technical scheme is that the method further comprises the step b: and (3) purifying the adsorbent obtained in the step (II), wherein the purifying treatment comprises water washing.

From the above, the invention grafts the polyanion through acidolysis reaction, the residual polyanion after the reaction can be dissolved in water, the salt produced by acidolysis such as sodium chloride and the like can also be dissolved in water, the purification of the adsorbent can be realized through water washing, the treatment steps are simple, the effect is good, and the product safety is high.

To achieve the second object of the present invention, there is provided a perfusion apparatus including an adsorbent prepared by the method for preparing an adsorbent for extracorporeal circulation of blood to remove LDL according to any one of the above aspects. The adsorbent prepared by the invention can be used for medical equipment for extracorporeal circulation of blood, namely an absorber or a perfusion device and the like.

Detailed Description

The adsorbent of the present invention and the method for preparing the same are further described below with reference to examples.

Example 1

The adsorbent was prepared in this example using the following steps:

step 1: the carrier resin containing epoxy groups is subjected to a ring opening reaction with an amine. The method comprises the following steps: 20mL of the poly glycidyl methacrylate resin is taken and mixed with an amination agent, and stirred and reacted for 1.5 hours at 50 ℃ to lead the epoxy groups on the carrier resin to have ring opening reaction. Wherein, the volume ratio of the polymethyl glycidyl acrylate resin to the amination agent is 1: (1-5); the amination agent is a mixture of lower amine (comprising ethylenediamine and the like) and water, and the volume ratio of the lower amine to the water is 1: (1-3); after the ring-opening reaction is finished, the obtained resin is washed by water until the effluent liquid is neutral.

Step 2: the product of step 1 is reacted with epichlorohydrin to graft hydrocarbon chlorine. The method comprises the following steps: adding a mixture of 10mL of epichlorohydrin and 20mL of water into the resin obtained in the step 1, stirring and reacting for 3h at 45 ℃ to ensure that the carrier resin carries a large amount of hydrocarbon chlorine groups, filtering out residual epichlorohydrin after the reaction is finished, and washing the resin with a small amount of alcohol and water.

Step 3: and (3) carrying out acidolysis reaction on the product obtained in the step (2) and polyacrylic acid to obtain the adsorbent. The method comprises the following steps: and (3) adding 20mL of polyacrylic acid aqueous solution (the molecular weight of polyacrylic acid is 25000) into the resin obtained in the step (2), and stirring and reacting for 4 hours at 45 ℃ to obtain the adsorbent. Wherein the mass ratio of polyacrylic acid to water is 1.5:5, a step of; the pH of polyacrylic acid aqueous solution is adjusted to 7-8 by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

Plasma adsorption performance evaluation:

10.0mL of high-fat plasma is added into 1.0mL of the adsorbent prepared in the example 1, the adsorbent is subjected to shaking adsorption for 2 hours at the constant temperature of 37 ℃, the supernatant is taken to detect the concentration of low-density lipoprotein (LDL), total Cholesterol (TC) and Triglyceride (TG), and a blank control experiment is carried out, and the detection result is shown in Table 1.

TABLE 1 results of detection of adsorption Performance of adsorbents prepared in example 1

Example 2

The adsorbent was prepared in this example using the following steps:

step 1: the cellulose carrier is epoxidised and then undergoes a ring opening reaction with an amine. The method comprises the following steps: taking 20mL of cellulose spheres, performing epoxidation by using epichlorohydrin or 1, 4-butanediol diglycidyl ether, controlling the pH value to be 8-11, and stirring and reacting for 3 hours at 30 ℃ to obtain the carrier after epoxidation. After the carrier is cleaned, the carrier is mixed with an amination agent and stirred for reaction for 1.5 hours at 50 ℃ to lead the epoxy groups on the carrier to carry out ring opening reaction. Wherein, the volume ratio of the carrier to the amination agent is 1: (1-5); the amination agent is a mixture of lower amine (including ammonia water, ethylenediamine and the like) and water, and the volume ratio of the lower amine to the water is 1: (1-3); after the ring-opening reaction is finished, the obtained product is washed by water until the effluent liquid is neutral.

Step 2: the product of step 1 is reacted with epichlorohydrin to graft hydrocarbon chlorine. The method comprises the following steps: and (2) adding a mixture of 10mL of epichlorohydrin and 20mL of water into the product obtained in the step (1), stirring and reacting for 3h at 45 ℃ to ensure that the carrier is provided with a large amount of hydrocarbon chlorine groups, filtering out residual epichlorohydrin after the reaction is finished, and washing the carrier with a small amount of alcohol and water.

Step 3: and (3) carrying out acidolysis reaction on the product obtained in the step (2) and polyacrylic acid to obtain the adsorbent. The method comprises the following steps: and (3) adding 20mL of polyacrylic acid aqueous solution into the product obtained in the step (2), and stirring at 45 ℃ for reaction for 4 hours to obtain the adsorbent. Wherein the mass ratio of polyacrylic acid to water is 1.5:5, a step of; the pH of polyacrylic acid aqueous solution is adjusted to 7-8 by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

Plasma adsorption performance evaluation:

10.0mL of high-fat plasma is added into 1.0mL of the adsorbent prepared in the example 2, the adsorbent is subjected to shaking adsorption for 2 hours at the constant temperature of 37 ℃, the supernatant is taken to detect the concentration of low-density lipoprotein (LDL), total Cholesterol (TC) and Triglyceride (TG), and a blank control experiment is carried out, and the detection results are shown in Table 2.

TABLE 2 detection results of adsorption Performance of adsorbents prepared in example 2

Example 3

This example uses a different chlorination reaction route than example 1 to prepare an adsorbent, comprising the steps of:

step 1: and adding 20mL of poly glycidyl methacrylate resin into a mixed solution of 2mL of concentrated hydrochloric acid and 80mL of acetone, stirring at 45 ℃ for reaction for 3h, and enabling epoxy groups on the carrier resin to carry out ring-opening reaction to obtain a carrier with hydrocarbon chloride groups, and washing the obtained resin with water until effluent liquid is neutral.

Step 2: and (3) carrying out acidolysis reaction on the product obtained in the step (1) and polyacrylic acid to obtain the adsorbent. Parameters such as reaction conditions refer to step 3 of example 1, specifically, 20mL of an aqueous solution of polyacrylic acid was added to the product obtained in step 1, and the mixture was stirred at 45℃for reaction for 4 hours to obtain an adsorbent. Wherein the mass ratio of polyacrylic acid to water is 1.5:5, a step of; the pH value of the solution is adjusted to 7-8 by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

Plasma adsorption performance evaluation:

10.0mL of high-fat plasma is added into 1.0mL of the adsorbent prepared in the example 3, the adsorbent is subjected to shaking adsorption for 2 hours at the constant temperature of 37 ℃, the supernatant is taken to detect the concentration of low-density lipoprotein (LDL), total Cholesterol (TC) and Triglyceride (TG), and a blank control experiment is carried out, and the detection result is shown in Table 3.

TABLE 3 detection results of adsorption Performance of adsorbents prepared in example 3

Comparing the adsorption performance test results of example 1 and example 3, it can be seen that the adsorption performance of the adsorbent of example 1 is superior to that of the adsorbent of example 3. The test was conducted in the same manner as in step 1 and step 2 of example 3 except that the polyglycidyl methacrylate resin was used for epoxidation of a carrier such as cellulose, and it was found that the adsorption performance of the adsorbent of example 1 was superior to that of the adsorbent obtained by the test in the same manner as described above. The reason for this is that the epoxy groups on the carrier resin react with hydrochloric acid to a low degree under conventional conditions, resulting in poor grafting effect in the post acidolysis reaction.

Example 4

The adsorbent was prepared in this example using the following steps:

Step 3: and (3) carrying out acidolysis reaction on the product obtained in the step (2) and polyacrylic acid to obtain the adsorbent. The method comprises the following steps: and (3) adding 20mL of polyacrylic acid aqueous solution into the resin obtained in the step (2), and stirring and reacting for a certain time at a certain temperature to obtain the adsorbent. Wherein the mass ratio of polyacrylic acid to water is 1.5:5, a step of; the pH of polyacrylic acid aqueous solution is adjusted to 7-8 by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

The adsorbents of examples 4-1 to 4-9 were prepared by adjusting the reaction temperature and reaction time in step 3 according to the reaction temperatures and reaction times shown in Table 4 below, and the results of the measured adsorption properties are shown in Table 4 below.

TABLE 4 detection results of adsorption Performance of adsorbents prepared in example 4

As can be seen from Table 4, the preparation method of the adsorbent for extracorporeal circulation removal LDL, TC, TG of blood according to the present invention can obtain an adsorbent with comparable adsorption performance by adjusting the reaction time when the acidolysis reaction temperature is between 35 ℃ and 55 ℃, and overcomes the defect of the prior art that the reaction time is long due to the fact that the reaction temperature is low in order to maintain the activity of EEDQ. From the convenience of production operation and production efficiency, the reaction temperature of 45 ℃ and the reaction time of 4 hours are selected as the optimal scheme of the scheme.

Example 5

The adsorbent was prepared in this example using the following steps:

step 1: the carrier resin containing epoxy groups is subjected to a ring opening reaction with an amine. The method comprises the following steps: mixing 20mL of poly (glycidyl methacrylate) resin with an amination agent, stirring at 50 ℃ for reaction for 1.5h, and enabling epoxy groups on the carrier resin to undergo ring opening reaction, wherein the volume ratio of the poly (glycidyl methacrylate) resin to the amination agent is 1: (1-5); the amination agent is a mixture of lower amine (comprising ethylenediamine and the like) and water, and the volume ratio of the lower amine to the water is 1: (1-3); after the ring-opening reaction is finished, the obtained resin is washed by water until the effluent liquid is neutral.

Step 3: and (3) carrying out acidolysis reaction on the product obtained in the step (2) and polyacrylic acid to obtain the adsorbent. The method comprises the following steps: and (3) adding 20mL of polyacrylic acid aqueous solution into the resin obtained in the step (2), and stirring and reacting for 4 hours at 45 ℃ to obtain the adsorbent. Wherein, the pH value of the polyacrylic acid solution is adjusted to 7-8 by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

The adsorbents of examples 5-1 to 5-6 were prepared by adjusting the concentration of the aqueous polyacrylic acid solution in step 3 according to the mass ratio of polyacrylic acid to water shown in Table 5, and the results of the measured adsorption properties are shown in Table 5 below.

TABLE 5 adsorption Performance test results of the adsorbents prepared in example 5

As can be seen from table 5, when the mass ratio of polyacrylic acid to water in the polyacrylic acid solution is lower than 1.5:5, the grafting amount of polyacrylic acid is low, excessive carboxyl genes on each polyacrylic acid molecular chain participate in the reaction and are consumed, and the adsorption performance of the adsorbent is poor; when the mass ratio of polyacrylic acid to water in the polyacrylic acid solution is higher than 1.5:5, the grafting amount of polyacrylic acid is higher, but the functional groups are too compact, so that the target substances adsorbed to the surface are not easy to diffuse inwards, and the adsorption performance is poor; when the mass ratio of polyacrylic acid to water in the polyacrylic acid solution is 1.5: and 5, the concentration is proper, so that the carboxyl groups participating in the reaction on each polyacrylic acid molecular chain are moderate, the excessive consumption of the carboxyl groups of the functional groups is avoided, and the functional groups are not excessively compact, thereby ensuring the adsorption performance of the polyacrylic acid molecular chain.

Example 6

The adsorbent was prepared in this example using the following steps:

Step 3: and (3) carrying out acidolysis reaction on the product obtained in the step (2) and polyacrylic acid to obtain the adsorbent. The method comprises the following steps: and (3) adding 20mL of polyacrylic acid aqueous solution into the resin obtained in the step (2), and stirring and reacting for 4 hours at 45 ℃ to obtain the adsorbent. Wherein the mass ratio of polyacrylic acid to water is 1.5:5, a step of; the pH of polyacrylic acid aqueous solution is adjusted by sodium hydride solution in advance; after the reaction, the adsorbent was washed with water.

The pH in step 3 was adjusted according to the pH values of the polyacrylic acid solutions listed in table 6, and adsorbents of examples 6-1 to 6-6 were prepared, and the measured adsorption performance results are shown in table 6 below.

TABLE 6 adsorption Performance test results of the adsorbents of example 6

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As can be seen from Table 6, when the pH of the polyacrylic acid solution was previously adjusted to 7 to 8 with sodium hydroxide, the carboxyl group in the polyacrylic acid was present in the form of COO ^－ The catalyst is easy to carry out acidolysis reaction with the hydrocarbon chlorine formed on the carrier in the step 2; when the pH is lower than 7, the carboxyl in the polyacrylic acid exists in a COOH form, and the hydroxyl in the form is not easy to undergo acidolysis reaction; when the pH is higher than 8, a certain amount of hydroxide ions exist in the solution, so that acidolysis reaction is easy to occur between the solution and the hydrocarbon chlorine, and the side reaction can be carried out between the solution and COO ^－ And (3) the acidolysis reaction competes, thereby affecting the grafting effect of the polyacrylic acid.

Example 7

And (3) taking 20mL of polyacrylic acid carrier resin, performing epoxidation by using propylene oxide or 1, 4-butanediol diglycidyl ether, controlling the pH value to be 8-11, and reacting for 3 hours at 30 ℃ to obtain the carrier after epoxidation. The epoxidized support was purified with purified water to pH neutral. Mixing the carrier after epoxidation with an amine agent according to the weight ratio of 1: (2-6) and reacting for 1.5h at 30 ℃ to aminate the carrier, wherein the aminating agent comprises ammonia water, ethylenediamine, 1, 2-propylenediamine or 1, 3-propylenediamine. The carrier was washed with purified water to a neutral pH. Mixing the aminated carrier with polyacrylic acid and EEDQ according to the mass ratio of 10:0.2:0.1, controlling the pH value to be 3-7, reacting for 12 hours at normal temperature, immobilizing polyacrylic acid (molecular weight is 5000-250000) ligand on a carrier, and cleaning and purifying the adsorbent to obtain the adsorbent. The LDL adsorption rate of this adsorbent was 62.5%. In the embodiment, EEDQ is used as a condensing agent, and the aminated carrier and polyacrylic acid are subjected to condensation reaction, so that the reaction is carried out at room temperature to avoid decomposition of the EEDQ, and the reaction speed is low and the reaction time is long.

From the above, the preparation method of the present invention does not adopt the method of accelerating the condensation of carboxyl and amino by condensing agents such as EEDQ in the prior art, but adopts halogenated epoxy compounds to connect the carrier containing amino and the polyanion, and realizes the grafting of the polyanion on the carrier through ring opening reaction and acidolysis reaction which are easier to react. Compared with the prior art, the invention can improve the production efficiency and the safety of products. In addition, experimental data show that the adsorbent obtained by the invention has good adsorption rate on low density lipoprotein, total cholesterol and triglyceride.

Finally, it should be emphasized that the above-described embodiments are merely exemplary of the present invention, and are not intended to limit the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing an adsorbent for extracorporeal circulation of blood for removing LDL, comprising the steps of:

step one: carrying out ring opening reaction on the carrier containing amino and halogenated epoxy compound to obtain intermediate product containing halogenated hydrocarbon group;

step two: and (3) carrying out acidolysis reaction on the intermediate product and polyanion to obtain the adsorbent.

2. The method for producing an adsorbent for removing LDL in extracorporeal blood circulation according to claim 1, wherein:

in the second step, the intermediate product is mixed with the aqueous solution of the polyanion and then subjected to acidolysis reaction; the mass ratio of the polyanion to water in the aqueous solution of the polyanion is (1.5-3): 5.

3. the method for producing an adsorbent for removing LDL in extracorporeal blood circulation according to claim 2, wherein: the mass ratio of the polyanion to water in the aqueous solution of the polyanion is 1.5:5.

4. the method for producing an adsorbent for removing LDL in extracorporeal blood circulation according to claim 1, wherein:

in the second step, the intermediate product is mixed with the aqueous solution of the polyanion and then subjected to acidolysis reaction; the pH of the aqueous solution of the polyanion is 7-8.

5. The method for producing an adsorbent for removing LDL in extracorporeal blood circulation according to claim 1, wherein:

in the second step, the reaction temperature of the acidolysis reaction is 35-60 ℃.

6. The method for producing an adsorbent for removing LDL in vitro blood circulation according to any one of claims 1 to 5, wherein:

the polyanion is at least one selected from polyacrylic acid, carbomer and dextran sulfate;

the halogenated epoxy compound is at least one selected from epichlorohydrin, methyl epichlorohydrin and 1, 2-epoxychlorobutane.

7. The method for producing an adsorbent for removing LDL in vitro blood circulation according to any one of claims 1 to 5, wherein:

in the first step, the carrier containing amino group and the halogenated epoxy compound are mixed with water and then subjected to ring opening reaction; the volume of water is 2 to 5 times the volume of the halogenated epoxy compound.

8. The method for producing an adsorbent for removing LDL in vitro blood circulation according to any one of claims 1 to 5, wherein:

the method further comprises the following steps: the carrier containing epoxy groups is subjected to ring opening reaction with amine to prepare the carrier containing amino groups in the step one.

9. The method for producing an adsorbent for removing LDL from blood extracorporeal circulation according to claim 8, wherein:

in the step a, the carrier containing the epoxy group is selected from at least one of macroporous styrene resin containing the epoxy group, macroporous acrylic resin and large Kong Qianwei element spheres; the amine is at least one of ammonia water, ethylenediamine, 1, 2-propylene diamine and 1, 3-propylene diamine; the ring-opening reaction is carried out after the carrier containing epoxy groups, amine and water are mixed.

10. The method for producing an adsorbent for removing LDL in vitro blood circulation according to any one of claims 1 to 5, wherein:

the second step further comprises a step b: and (3) purifying the adsorbent obtained in the step (II), wherein the purifying treatment comprises water washing.

11. Perfusion apparatus, its characterized in that:

the perfusion apparatus includes the adsorbent prepared by the method for preparing the adsorbent for extracorporeal blood circulation to remove LDL according to any one of claims 1 to 10.