Adsorption material for blood perfusion and preparation method thereof

Abstract

The invention relates to an adsorbing material for blood perfusion and a preparation method thereof. The preparation method comprises the following steps: providing a hydrophobic porous carrier which comprises styrene-divinyl benzene copolymer microspheres, and introducing a coordination group on the surface of the hydrophobic porous carrier through chemical modification. The average pore size of the hydrophobic porous carrier is 2-50 nm, and the specific surface area of the hydrophobic porous carrier is greater than or equal to 500 m < 2 > / g; the coordination group is selected from at least one of iminodiacetic acid (IDA), ethylenediaminetetraacetic acid (EDTA) and N, N, N-tricarboxymethyl ethylenediamine (TED). The prepared adsorption material is high in A beta removal capacity, has certain removal capacity on heavy metal ions, is good in blood compatibility and excellent in anticoagulation performance, and is an ideal material for preparing a blood perfusion device.

Description

Technical Field

[0001] The present invention relates to the technical field of adsorbents, in particular to an adsorption material for blood perfusion and a preparation method thereof. Background Art

[0002] Alzheimer's disease (AD) is a chronic neurodegenerative disorder characterized by progressive cognitive impairment, memory loss, and behavioral abnormalities. Its pathological hallmarks include senile plaques formed by the deposition of β-amyloid (Aβ), neurofibrillary tangles (NFTs) caused by hyperphosphorylated tau protein, neuronal loss, and chronic neuroinflammation. Recent studies have gradually revealed the important role of cytokines and Aβ in the pathogenesis of AD.

[0003] Aβ is a peptide produced by the cleavage of amyloid precursor protein (APP) by β-secretase and γ-secretase. In 1984, scientists first isolated Aβ from the brains of AD patients and found its deposition in senile plaques. It is reported that the concentration of iron ions in the cortex is proportional to the probability of plaque formation. As the amyloid content increases, the iron ion concentration increases, and plaque formation accelerates. Studies have shown that metal ions (especially Cu 2+ ) content is much higher than that of normal people. Copper ions may play an important role in the pathogenesis of AD. Excessive accumulation of copper ions may lead to neuronal loss and cell death, thereby further accelerating the progression of the disease. Therefore, researchers believe that reducing the accumulation of copper ions in the brain may become a new treatment strategy, which is expected to bring better therapeutic effects to Alzheimer's patients. The impaired function of amyloid precursor protein in AD can trigger intracellular and extracellular Fe 2+ and Cu 2+ Elevated levels and extracellular Cu 2+ Reduce, thereby promoting its accumulation. Excess Fe 2+ and Cu 2+ Increased oxidative stress occurs by generating ROS. Furthermore, iron, copper, and zinc have high binding affinity for Aβ, promoting its aggregation. Aβ oligomers are neurotoxic, disrupting synaptic function, inducing oxidative stress, and leading to neuronal death. Normally, Aβ is cleared through microglial phagocytosis, enzymatic degradation (such as neprilysin), and efflux transport (such as that mediated by LRP1). However, these clearance mechanisms are impaired in AD, leading to Aβ accumulation.

[0004] In the 1990s, studies discovered the presence of activated microglia and astrocytes in the brains of patients with Alzheimer's disease, suggesting a role for neuroinflammation in AD. Upon activation, microglia and astrocytes release large amounts of cytokines (such as IL-1β, IL-6, and TNF-α) and chemokines, triggering chronic neuroinflammation. While this inflammatory response may have protective effects in the early stages (such as clearing Aβ), it can exacerbate neuronal damage and synaptic dysfunction in the chronic stage.

[0005] Aβ deposition can activate microglia and astrocytes, prompting them to release proinflammatory cytokines, triggering neuroinflammation. Proinflammatory cytokines (such as IL-1β and TNF-α) promote abnormal processing of APP, increasing Aβ production while inhibiting Aβ clearance, forming a positive feedback loop. Chronic inflammation leads to neuronal damage, synaptic dysfunction, and cognitive decline.

[0006] Most existing blood perfusion products use polystyrene divinylbenzene as an adsorbent, which has poor blood compatibility and a low adsorption rate for cytokines. It cannot be used to adsorb and remove cytokines, Aβ and metal ions, and therefore cannot meet the needs of AD patients. Summary of the Invention

[0007] The purpose of the present invention is to disclose an adsorption material for blood perfusion and a preparation method thereof, so as to solve one or more technical problems existing in the prior art and provide at least one beneficial option or create conditions.

[0008] A first aspect of the present invention is to provide a method for preparing an adsorption material.

[0009] A second aspect of the present invention is to provide an adsorption material.

[0010] The third aspect of the present invention is to provide use of the adsorption material according to the second aspect of the present invention.

[0011] The material preparation method described in the first aspect of the present invention comprises the following steps: A hydrophobic porous carrier is provided, comprising styrene-divinylbenzene copolymer microspheres, wherein a coordination group is introduced on the surface of the hydrophobic porous carrier by chemical modification. The hydrophobic porous carrier has an average pore size of 2 to 50 nm and a specific surface area of ​​≥500 m 2 / g; the coordination group is selected from at least one of iminodiacetic acid (IDA), ethylenediaminetetraacetic acid (EDTA), and N,N,N-tricarboxymethylethylenediamine (TED).

[0012] In further application embodiments, the chemical modification can be achieved through three synthetic pathways.

[0013] The first synthetic route: 1-1) epoxidizing the double bonds of the copolymer microspheres to form epoxy groups; 1-2) reacting the epoxy group with an aminating agent at 50-90° C., wherein the aminating agent is selected from at least one of iminodiacetic acid and N,N,N-tricarboxymethylethylenediamine.

[0014] The second synthetic route: 2-1) epoxidizing the double bonds of the copolymer microspheres to form epoxy groups; 2-2) reacting the epoxy group with an aminating agent at 50-90° C. to form a first intermediate containing amino groups on the surface, wherein the aminating agent is selected from polyethyleneimine, polylysine, C2-C 12 at least one of an alkyl diamine; 2-3) performing a coupling reaction on the carboxylic acid ligand and the first intermediate in the presence of a condensing agent, wherein the carboxylic acid ligand is selected from at least one of iminodiacetic acid and ethylenediaminetetraacetic acid, and the condensing agent is selected from at least one of carbodiimides and N-hydroxysuccinimide.

[0015] The third synthetic route: 3-1) Chloromethylating the benzene rings of the copolymer microspheres to form a chloromethylated intermediate; 3-2) reacting the chloromethylated intermediate with an aminating agent at 50-90° C. to form a second intermediate containing amino groups on the surface, wherein the aminating agent is selected from polyethyleneimine, polylysine, C2-C 12 at least one of an alkyl diamine; 3-3) reacting the second intermediate with a carboxylic acid ligand under alkaline conditions, wherein the carboxylic acid ligand is at least one selected from the group consisting of iminodiacetic acid and N,N,N-tricarboxymethylethylenediamine.

[0016] In a further application embodiment, the C2-C 12 The alkyldiamine is at least one selected from ethylenediamine and hexamethylenediamine.

[0017] In a further application embodiment, the cross-linking degree of the copolymer microspheres is 1-20%, preferably 5-15%.

[0018] In a further application embodiment, the average particle size of the copolymer microspheres is 10-1000 μm, preferably 300-800 μm.

[0019] In a further application embodiment, the chemical modification further comprises adding a phase transfer catalyst into the reaction system, wherein the phase transfer catalyst is selected from at least one of quaternary ammonium salts and crown ethers.

[0020] In a further application embodiment, the loading amount of the ligand group is 0.1-5 mmol / g carrier.

[0021] The adsorption material of the second aspect of the present invention is prepared by the above-mentioned preparation method, and the adsorption capacity of the adsorption material for transition metal ions is ≥5 mg / g. The adsorption material can remove cytokines and Aβ by utilizing the van der Waals force of the copolymer microspheres, and the obtained coordination group has electrostatic and chelating effects, which can remove Fe 2+ 、Cu 2+ ion.

[0022] The third aspect of the present invention relates to the use of the adsorbent material in the preparation of a hemoperfusion device. Hemoperfusion technology is commonly used clinically to remove toxins and pathogenic substances from the blood, thereby purifying the blood and alleviating and treating diseases. During treatment, the removal of cytokines (primarily IL-6), Aβ, and metal ions is a research challenge and an important evaluation criterion for hemoperfusion devices. The adsorbent material of the present invention has a high Aβ removal capacity, a moderate heavy metal ion removal capacity, good blood compatibility, and excellent anticoagulant properties, making it an ideal material for the preparation of hemoperfusion devices. DETAILED DESCRIPTION

[0023] In the description of the present invention, if there is a description of first, second, etc., it is only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0024] The terms "preferably," "more preferably," and the like, used herein refer to embodiments of the present invention that may provide certain benefits under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, nor is it intended to exclude other embodiments from the scope of the present invention.

[0025] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein are understood to include any and all subranges subsumed therein.

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts are within the scope of the present invention.

[0027] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0028] Example 1 The adsorption material is prepared by a first synthetic route.

[0029] 1-1) Epoxidation: Take an appropriate amount of polystyrene divinylbenzene resin (prepared by suspension polymerization, pore size 2~50 nm, specific surface area>500m 2 / g), add 40 mL of dichloroethane and soak overnight. Then add 20 mL of a 1% solution of m-chloroperbenzoic acid in dichloroethane. Stir in an ice-water bath for 24 hours at 150-250 rpm. After the reaction is complete, rinse with alcohol and purified water.

[0030] 1-2) Fixed IDA: Add 40 mL of 1% IDA solution to the epoxidized resin and shake at 60-80°C for 24 hours. After the reaction, wash with alcohol and purified water.

[0031] The adsorption material S1 is obtained.

[0032] Example 2 The adsorption material is prepared by a first synthetic route.

[0033] 1-1) Epoxidation: Take an appropriate amount of polystyrene divinylbenzene resin (prepared by suspension polymerization, pore size 2~50 nm, specific surface area>500m 2 / g (hereinafter referred to as "adsorbent material S0"), add 40 mL of dichloroethane and soak overnight. Then add 20 mL of a 1% solution of m-chloroperbenzoic acid in dichloroethane. Stir in an ice-water bath for 24 hours at 150-250 rpm. After the reaction is complete, rinse with alcohol and purified water.

[0034] 1-2) Immobilized TDA: Add 40 mL of 1% TDA solution to the epoxidized resin and shake at 60-80°C for 24 hours. After the reaction, wash with alcohol and purified water.

[0035] The adsorption material S2 is obtained.

[0036] Example 3 The adsorption material was prepared by a second synthetic route.

[0037] 3-1) Epoxidation: Take the adsorbent material S0 and soak it in 40 mL of dichloroethane overnight. Then add 20 mL of a 1% solution of m-chloroperbenzoic acid in dichloroethane. Stir in an ice-water bath for 24 hours at 150-250 rpm. After the reaction is complete, rinse with alcohol and purified water.

[0038] 3-2) Amination: The epoxidized resin was added to 40 mL of a 2% aqueous polyethyleneimine solution and shaken at 60-80°C for 24 hours. After the reaction, the mixture was washed with alcohol and purified water to obtain the first intermediate.

[0039] 3-3) Condensed IDA: Add 40 mL of a 1% IDA solution to the first intermediate, adjust the pH to 4.8, add 0.6 g of N-hydroxysuccinimide (NHS), and then slowly add 0.4 g of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC). Shake and react at 25°C for 5 hours. After the reaction, sterilize the solution with alcohol and water for injection.

[0040] The adsorption material S3 is obtained.

[0041] Example 4 The adsorption material was prepared by a second synthetic route.

[0042] 4-1) Epoxidation: Take the adsorbent material S0 and soak it in 40 mL of dichloroethane overnight. Then add 20 mL of a 1% solution of m-chloroperbenzoic acid in dichloroethane. Stir in an ice-water bath for 24 hours at 150-250 rpm. After the reaction is complete, rinse with alcohol and purified water.

[0043] 4-2) Amination: The epoxidized resin was added to 40 mL of a 2% aqueous polyethyleneimine solution and shaken at 60-80°C for 24 hours. After the reaction, the mixture was washed with alcohol and purified water to obtain the first intermediate.

[0044] 4-3) Condensation of EDTA: Add 40 mL of 2.2% EDTA solution to the first intermediate, adjust the pH to 4.8, add 0.6 g of NHS, and then slowly add 0.4 g of EDC. Shake and react at 25°C for 5 hours. After the reaction is complete, sterilize with alcohol and water for injection.

[0045] The adsorption material S4 is obtained.

[0046] Example 5 The adsorption material was prepared by the third synthetic route.

[0047] 5-1) Chloromethylation: Take 10 g of adsorbent material S0 and add 40 mL of chloromethyl ether to swell for 2 hours. Then add 6 g of zinc chloride, raise the temperature to 42°C, and react for 16 hours to obtain the chloromethylated resin. After the reaction is complete, wash with alcohol, purified water, or a similar method.

[0048] 5-2) Amination: Take the chloromethylated resin, add 40 mL of a 2% polylysine aqueous solution, and shake at 60-80°C for 24 hours. After the reaction is complete, wash with alcohol and purified water to obtain the second intermediate.

[0049] 5-3) Fixed IDA: The second intermediate was added to 40 mL of alkaline solution and 0.4 g of IDA. After the reaction was complete, the mixture was sterilized by washing with alcohol and water for injection.

[0050] The adsorption material S5 is obtained.

[0051] Example 6 The adsorption material was prepared by the third synthetic route.

[0052] 6-1) Chloromethylation: Take 10 g of adsorbent material S0 and add 40 mL of chloromethyl ether to swell for 2 hours. Then add 6 g of zinc chloride, raise the temperature to 42°C, and react for 16 hours to obtain the chloromethylated resin. After the reaction is complete, wash with alcohol, purified water, or a similar method.

[0053] 6-2) Amination: Take the chloromethylated resin, add 40 mL of a 2% polylysine aqueous solution, and shake at 60-80°C for 24 hours. After the reaction is complete, wash with alcohol and purified water to obtain the second intermediate.

[0054] 6-3) Immobilized TDA: The second intermediate was added to 40 mL of alkaline solution and 0.4 g of TDA. After the reaction was complete, the mixture was sterilized by washing with alcohol and water for injection.

[0055] The adsorption material S6 is obtained.

[0056] Example 7: Heavy metal ion adsorption experiment Take a test tube, add a certain amount of adsorbent material to each tube, then add 1.5 mL of a solution containing heavy metal ions. Oscillating adsorption is carried out at 37°C and a rate of 100 ± 10 rpm for 2 hours. The concentration of the corresponding ion is then measured, and the removal rate of the adsorbent material is calculated. The results are shown in Table 1.

[0057] Table 1. Adsorption of heavy metal Fe by adsorption materials 2+ 、Cu 2+ Adsorption performance and removal rate

[0058] It can be seen from Table 1 that the adsorbent material has a certain ability to remove heavy metal ions. After being immobilized with ligands IDA, EDTA or TED, the adsorption of heavy metal Fe 2+ 、Cu 2+ The removal rates of the adsorbent materials S2 and S6 were improved, but in comparison, the removal efficiencies of the adsorbent materials S2 and S6 loaded with TDA ligands were lower than those of the adsorbent materials loaded with IDA ligands and EDTA ligands, so they were not involved in subsequent experiments.

[0059] Example 8: Cytokine adsorption experiment Take a pyrogen-free test tube and add 0.25 mL of the prepared adsorbent material. Then add 2.5 mL of plasma containing 478.00 pg / mL of cytokines interleukin-6 (IL-6) and 716.85 pg / mL of tumor necrosis factor α (TNF-α). Oscillating adsorption was carried out at 37°C and a shaking rate of 100±10 rpm for 2 hours. The concentrations of IL-6 and TNF-α were then measured, and the clearance rate of the adsorbent material was calculated.

[0060] Table 2. Clearance rate of cytokines IL-6 and TNF-α by adsorption materials

[0061] It can be found that the adsorption material has a certain ability to clear cytokines IL-6 and TNF-α.

[0062] Example 9: Adsorption of β-amyloid protein Take a test tube, add a certain amount of adsorption material, then add 1.5 mL of Aβ solution, and shake and adsorb for 2 hours at a temperature of 37°C and a shaking rate of 100±10 rpm. Then detect the concentration of Aβ and calculate its clearance rate by the adsorption material.

[0063] Table 3. Adsorption performance and clearance rate of adsorbent materials for Aβ

[0064] Table 3 shows that polystyrene divinylbenzene resin itself has a certain ability to clear Aβ. After modification, the clearance rate of Aβ-amyloid protein is improved.

[0065] Example 10, blood compatibility experiment One gram of each adsorbent material was soaked in saline for 10 hours before being loaded into a small cartridge. Then, 10 mL of heparinized rabbit whole blood was injected via syringe at a flow rate of 50 mL / minute for 2 hours. An empty cartridge was also added for a control experiment. Changes in blood components before and after perfusion were measured. The results showed that the concentrations of major blood components, such as red blood cells, remained minimal before and after perfusion, with the percentage change within 15%. This demonstrates that this series of adsorbents has good hemocompatibility and has promising applications in whole blood perfusion.

[0066] Example 11: Anticoagulant performance evaluation experiment Recalcification time is used to evaluate anticoagulant performance. A longer recalcification time indicates a better anticoagulant effect. Comparing the recalcification time of the adsorbent material before and after modification, the table below shows that the adsorbent material S4 obtained in Example 4 exhibits better anticoagulant efficacy than conventional adsorbents.

[0067] Table 4. Results of the recalcification time experiment

[0068] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

Claims

1. A method for preparing an adsorption material, characterized in that: Including steps: A hydrophobic porous carrier is provided, wherein the hydrophobic porous carrier comprises styrene-divinylbenzene copolymer microspheres, wherein the hydrophobic porous carrier has an average pore size of 2 to 50 nm and a specific surface area of ​​≥500 m 2 / g; A coordination group is introduced on the surface of the hydrophobic porous carrier by chemical modification, wherein the coordination group is selected from at least one of iminodiacetic acid, ethylenediaminetetraacetic acid, and N,N,N-tricarboxymethylethylenediamine.

2. The preparation method according to claim 1, characterized in that The chemical modification includes: 1-1) epoxidizing the double bonds of the copolymer microspheres to form epoxy groups; 1-2) reacting the epoxy group with an aminating agent at 50-90° C., wherein the aminating agent is selected from at least one of iminodiacetic acid and N,N,N-tricarboxymethylethylenediamine.

3. The preparation method according to claim 1, characterized in that The chemical modification includes: 2-1) epoxidizing the double bonds of the copolymer microspheres to form epoxy groups; 2-2) reacting the epoxy group with an aminating agent at 50-90° C. to form a first intermediate containing amino groups on the surface, wherein the aminating agent is selected from polyethyleneimine, polylysine, C2-C 12 at least one of an alkyl diamine; 2-3) performing a coupling reaction on the carboxylic acid ligand and the first intermediate in the presence of a condensing agent, wherein the carboxylic acid ligand is selected from at least one of iminodiacetic acid and ethylenediaminetetraacetic acid, and the condensing agent is selected from at least one of carbodiimides and N-hydroxysuccinimide.

4. The preparation method according to claim 1, characterized in that The chemical modification includes: 3-1) Chloromethylating the benzene rings of the copolymer microspheres to form a chloromethylated intermediate; 3-2) reacting the chloromethylated intermediate with an aminating agent at 50-90° C. to form a second intermediate containing amino groups on the surface, wherein the aminating agent is selected from polyethyleneimine, polylysine, C2-C 12 at least one of an alkyl diamine; 3-3) reacting the second intermediate with a carboxylic acid ligand under alkaline conditions, wherein the carboxylic acid ligand is at least one selected from the group consisting of iminodiacetic acid and N,N,N-tricarboxymethylethylenediamine.

5. The preparation method according to claim 3 or 4, characterized in that The C2-C 12 The alkyldiamine is at least one selected from ethylenediamine and hexamethylenediamine.

6. The preparation method according to any one of claims 1 to 4, characterized in that The crosslinking degree of the copolymer microspheres is 1-20%, and the average particle size is 10-1000 μm.

7. The preparation method according to any one of claims 1 to 4, characterized in that The chemical modification further comprises adding a phase transfer catalyst into the reaction system, wherein the phase transfer catalyst is selected from at least one of quaternary ammonium salts and crown ethers.

8. The preparation method according to any one of claims 1 to 4, characterized in that The loading amount of the coordination group is 0.1-5 mmol / g carrier.

9. An adsorption material, characterized in that Prepared by the preparation method according to any one of claims 1 to 8, the adsorption capacity of the adsorption material for transition metal ions is ≥50 mg / g.

10. Use of the adsorption material according to claim 9 in preparing a hemoperfusion product.

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