A protein-bound toxin adsorbent, and a method for preparing and using the same

Abstract

The application provides a protein-bound toxin adsorbent and a preparation method and application thereof, the protein-bound toxin adsorbent takes super-high crosslinking styrene-divinylbenzene resin as a carrier, the carrier is loaded with polyethyleneimine, the polyethyleneimine is connected with an adsorption ligand, and the adsorption ligand comprises at least one of alpha-methyl-4-(2-methylpropyl) phenylacetic acid, oleic acid and linoleic acid. The protein-bound toxin adsorbent of the application makes the protein-bound toxin in a free state through the action of the adsorption ligand, and simultaneously removes the protein-bound toxin in the free state through the adsorption action of the super-high crosslinking styrene-divinylbenzene resin and the electrostatic combination of the polyethyleneimine, so that the removal rate of the protein-bound toxin is improved.

Description

Technical Field

[0001] This invention relates to the field of blood purification technology, and more specifically, to a protein-bound toxin adsorbent, its preparation method, and its application. Background Technology

[0002] Uremia, also known as end-stage renal disease, is a clinical syndrome of the end stage of chronic renal failure. Uremic patients have higher concentrations of approximately two hundred substances in their bodies than normal individuals. Based on their biochemical properties, these substances are mainly classified into three categories: small-molecule water-soluble solutes, medium- and large-molecule toxins, and protein-bound toxins.

[0003] Protein-bound toxins (PBUTs) typically have a relative molecular mass below 500. Although their molecular weight is small, PBUTs readily bind to serum proteins, forming larger protein-bound structures that accumulate in uremia patients. This accumulation can impair the function of various systems in uremia patients, leading to complications. Numerous studies have shown that PBUTs are involved in the progression of chronic renal failure and are closely related to renal interstitial fibrosis and cardiovascular complications of CKD. Therefore, it is necessary to eliminate PBUTs from the bodies of uremia patients.

[0004] Traditional techniques such as dialysis, filtration, dialysis-filtration, and high-flux dialysis have limited ability to remove protein-bound toxins. Existing blood purification systems that combine adsorbents and dialyzers have significantly improved the removal of protein-bound toxins compared to traditional dialysis methods, but the removal efficiency of these systems is still not ideal. Furthermore, the adsorbents in existing blood purification systems lack selectivity in adsorbing protein-bound toxins. Summary of the Invention

[0005] The present invention aims to solve the problem that the existing technology has poor clearance effect on protein-bound toxins.

[0006] To address the aforementioned problems, the first aspect of this invention provides a protein-bound toxin adsorbent, wherein the protein-bound toxin adsorbent uses ultra-high cross-linked styrene-divinylbenzene resin as a carrier, and polyethyleneimine is immobilized on the carrier. The polyethyleneimine is connected to an adsorption ligand, and the adsorption ligand includes at least one selected from α-methyl-4-(2-methylpropyl)phenylacetic acid, oleic acid, and linoleic acid.

[0007] Furthermore, the ultra-high crosslinked styrene-divinylbenzene resin is prepared by crosslinking a low crosslinked styrene-divinylbenzene copolymer, and the polyethyleneimine is introduced during the crosslinking reaction of the low crosslinked styrene-divinylbenzene copolymer. The adsorption ligand is attached to the polyethyleneimine by electrostatic adsorption, and the loading of the polyethyleneimine ranges from 0.1 mmol / g to 1 mmol / g.

[0008] A second aspect of the present invention provides a method for preparing a protein-binding toxin adsorbent, used to prepare the protein-binding toxin adsorbent described in the first aspect, the method comprising:

[0009] Preparation of low-crosslinked styrene-divinylbenzene copolymer;

[0010] The low-crosslinked styrene-divinylbenzene copolymer undergoes a crosslinking reaction to prepare a highly crosslinked styrene-divinylbenzene resin, wherein polyethyleneimine is immobilized on the highly crosslinked styrene-divinylbenzene resin, and the polyethyleneimine is immobilized onto the highly crosslinked styrene-divinylbenzene resin during the crosslinking reaction of the low-crosslinked styrene-divinylbenzene copolymer.

[0011] The ultra-high cross-linked styrene-divinylbenzene resin was immersed in an adsorption ligand solution. After immersion, the adsorption ligand was attached to the polyethyleneimine to obtain a protein-bound toxin adsorbent.

[0012] Further, the low-crosslinked styrene-divinylbenzene copolymer undergoes a crosslinking reaction to prepare a highly crosslinked styrene-divinylbenzene resin, comprising:

[0013] The low crosslinked styrene-divinylbenzene copolymer was subjected to a chloromethylation reaction, and polyethyleneimine was added during the chloromethylation reaction to obtain a low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

[0014] The low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine was subjected to a Friedel-Crafts reaction to obtain a highly crosslinked styrene-divinylbenzene resin.

[0015] Further, the step of subjecting the low-crosslinked styrene-divinylbenzene copolymer to a chloromethylation reaction, and adding polyethyleneimine during the chloromethylation reaction to obtain a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine, comprises:

[0016] After swelling in dichloroethane, the low-crosslinked styrene-divinylbenzene copolymer, chloromethyl ether, and anhydrous ferric chloride are refluxed at 50°C to 60°C for 2 to 4 hours. Then, the temperature is raised to 80°C to 100°C and refluxed for another 5 to 24 hours. Simultaneously, polyethyleneimine is added during the heating process to introduce the polyethyleneimine onto the low-crosslinked styrene-divinylbenzene copolymer, thus obtaining a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

[0017] Further, the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the chloromethyl ether is 1:4 to 1:6, the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the anhydrous ferric chloride is 1:0.5 to 1:1.5, and the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the polyethyleneimine is 20:5 to 20:20.

[0018] Further, the step of subjecting the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine to a Friedel-Crafts reaction to obtain a highly crosslinked styrene-divinylbenzene resin comprises:

[0019] The low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine is mixed with nitrobenzene and allowed to stand at 35°C to 45°C for 4 to 5 hours. Zinc chloride is then added under stirring to carry out a Friedel-Crafts reaction to form a highly crosslinked styrene-divinylbenzene resin. The Friedel-Crafts reaction is carried out at a temperature of 110°C to 130°C for 8 to 16 hours.

[0020] Further, the process involves immersing the ultra-highly cross-linked styrene-divinylbenzene resin in an adsorption ligand solution. After immersion, the adsorption ligand is attached to the polyethyleneimine to obtain a protein-binding toxin adsorbent, comprising:

[0021] The ultra-highly crosslinked styrene-divinylbenzene resin is immersed in an alkaline solution until the pH value of the alkaline solution remains unchanged to obtain the transformed ultra-highly crosslinked styrene-divinylbenzene resin. After the transformed ultra-highly crosslinked styrene-divinylbenzene resin is cleaned and dried, it is immersed in an adsorbent ligand solution. After immersion, it is cleaned to obtain a protein-binding toxin adsorbent, wherein the mass percentage of adsorbent ligand in the adsorbent ligand solution is 10% to 40%.

[0022] Furthermore, the transformed ultra-high crosslinked styrene-divinylbenzene resin is cleaned with deionized water after being soaked in the adsorbent ligand.

[0023] A third aspect of the present invention provides a blood perfusion device comprising a protein-binding toxin adsorbent as described in the first aspect, or a protein-binding toxin adsorbent prepared by the preparation method described in the second aspect.

[0024] The protein-binding toxin adsorbent of this invention uses ultra-high cross-linked styrene-divinylbenzene resin as a carrier to immobilize polyethyleneimine. The polyethyleneimine is linked to adsorption ligands, which carry a negative charge. When this protein-binding toxin adsorbent comes into contact with blood, the adsorption ligands detach from the adsorbent and enter the bloodstream. The adsorption ligands have the same albumin-binding sites as p-cresol sulfate and indophenol sulfate, enabling them to compete with protein-binding toxins for adsorption onto albumin. Furthermore, the binding force between the adsorption ligands and albumin is stronger than that between the protein-binding toxin and albumin, causing the protein-binding toxin to detach from the albumin and thus preventing the protein-binding toxin from entering the bloodstream. Protein-bound toxins exist in a free state, facilitating adsorption by ultra-highly cross-linked styrene-divinylbenzene resin and the polyethyleneimine immobilized thereon, thereby improving the clearance rate of protein-bound toxins. Simultaneously, using ultra-highly cross-linked styrene-divinylbenzene resin as a carrier, its abundant porous structure allows for the adsorption of medium-to-large molecules and small-molecule protein-bound toxins from uremic toxins. The positively charged polyethyleneimine immobilized on the ultra-highly cross-linked styrene-divinylbenzene resin can electrostatically bind to free protein-bound toxins in the blood, thus clearing them. The protein-bound toxin adsorbent of this invention, through the adsorption of ligands, keeps protein-bound toxins in a free state, while simultaneously clearing these free-state protein-bound toxins through the adsorption of ultra-highly cross-linked styrene-divinylbenzene resin and the electrostatic binding of polyethyleneimine, thereby improving the clearance rate of protein-bound toxins.

[0025] The method for preparing the protein-bound toxin adsorbent of this invention involves first preparing a low-crosslinked styrene-divinylbenzene copolymer via suspension polymerization, and then preparing a highly crosslinked ethylene-divinylbenzene resin from the low-crosslinked styrene-divinylbenzene copolymer. This highly crosslinked ethylene-divinylbenzene resin possesses a rich porous structure, enabling it to adsorb medium- and large molecular weight substances and small protein-bound toxins in uremic toxins. Polyethyleneimine is added during the crosslinking reaction of the low-crosslinked styrene-divinylbenzene copolymer, and the chloromethyl groups generated during the reaction immobilize the polyethyleneimine onto the highly crosslinked styrene-divinylbenzene resin. This avoids the introduction of other reagents or substances, simplifying the reaction process, improving reaction efficiency, and preventing any impact on the adsorption performance of the highly crosslinked styrene-divinylbenzene resin. Subsequently, immersing the highly crosslinked styrene-divinylbenzene resin in an adsorption ligand solution allows the adsorption ligand to enter the porous structure of the resin, facilitating sufficient contact and binding between the adsorption ligand and the polyethyleneimine immobilized on the resin, thus promoting the binding of polyethyleneimine to the adsorption ligand. The preparation method provided by this invention simplifies the reaction steps, is simple to prepare, has mild reaction conditions, high safety during the reaction process, and low production cost, which is conducive to large-scale industrial production. Attached Figure Description

[0026] Figure 1 This is a process flow diagram for preparing protein-binding toxin adsorbents according to an embodiment of the present invention. Detailed Implementation

[0027] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0028] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0029] Furthermore, the terms "comprising," "including," "containing," and "having" are non-restrictive and can refer to the addition of other steps and components that do not affect the results. Unless otherwise specified, all materials, equipment, and reagents are commercially available.

[0030] Furthermore, although the present invention describes each step in the preparation process in the form of S110, S120, S130, etc., this description is only for ease of understanding. The forms such as S110, S120, S130 do not indicate a limitation on the order of the steps.

[0031] The first aspect of the embodiments of this application provides a protein-binding toxin adsorbent, which uses ultra-high cross-linked styrene-divinylbenzene resin as a carrier, on which polyethyleneimine is immobilized, and the polyethyleneimine is connected to an adsorption ligand, the adsorption ligand including at least one of α-methyl-4-(2-methylpropyl)phenylacetic acid, oleic acid and linoleic acid.

[0032] The protein-bound toxins include hippuric acid (HA), indole-3-acetic acid (IAA), indophenol sulfate (IS), p-cresol sulfate (PCS), and 3-carboxy-4-methyl-5-propyl-2-furanopropionic acid (CMPF). The protein-bound toxin adsorbent in this embodiment can adsorb the above-mentioned protein-bound toxins, especially IAA, IS, and PCS.

[0033] Specifically, α-methyl-4-(2-methylpropyl)phenylacetic acid, also known as ibuprofen, has a π-electron conjugated structure and a carboxyl group. Its carboxyl group dissociates under neutral pH conditions, carrying a single negative charge. This negatively charged carboxyl group can interact electrostatically with positively charged groups, and its benzene ring forms a hydrophobic interaction with the four leucine residues arranged in a ring in the center of the cavity. Furthermore, α-methyl-4-(2-methylpropyl)phenylacetic acid has the same albumin binding site as p-cresol sulfate and indophenol sulfate. With the aid of hydrophobic and electrostatic interactions, α-methyl-4-(2-methylpropyl)phenylacetic acid can compete with protein-bound toxins for adsorption onto albumin. Moreover, the binding force between α-methyl-4-(2-methylpropyl)phenylacetic acid and albumin is stronger than that between protein-bound toxins and albumin, thereby causing the protein-bound toxins to detach from albumin and become free, which facilitates the adsorption of protein-bound toxins by the adsorbent.

[0034] Oleic acid and linoleic acid both have carboxyl groups. The carboxyl groups dissociate under neutral pH conditions, carrying a single negative charge. The negatively charged carboxyl groups can interact electrostatically with positively charged groups. Both substances are hydrophobic and have the same albumin binding sites as cresol sulfate and indophenol sulfate. By attaching oleic acid and / or linoleic acid to the protein-bound toxin adsorbent, the hydrophobic and electrostatic interactions of oleic acid and / or linoleic acid can help the protein-bound toxin detach from albumin, making the protein-bound toxin free and facilitating its adsorption by the adsorbent.

[0035] Furthermore, α-methyl-4-(2-methylpropyl)phenylacetic acid is highly safe for human use, is metabolizable, and has relatively few toxic side effects; oleic acid has cis-trans isomers, which have a certain effect on softening blood vessels and play an important role in the metabolism of humans and animals; linoleic acid is an essential fatty acid for the human body, which can lower blood cholesterol and prevent atherosclerosis. Therefore, α-methyl-4-(2-methylpropyl)phenylacetic acid, oleic acid, and linoleic acid are relatively safe for use as protein-bound toxin adsorbents.

[0036] In this embodiment, the ultra-high crosslinked styrene-divinylbenzene resin is obtained by crosslinking a low crosslinked styrene-divinylbenzene copolymer. Polyethyleneimine is introduced during the crosslinking reaction of the low crosslinked styrene-divinylbenzene copolymer, and the adsorption ligand is attached to the polyethyleneimine by electrostatic adsorption.

[0037] Specifically, the ultra-highly crosslinked styrene-divinylbenzene resin is prepared by chloromethylation and Friedel-Crafts crosslinking of low-crosslinked styrene-divinylbenzene copolymer. Polyethyleneimine is introduced during the chloromethylation reaction of the low-crosslinked styrene-divinylbenzene copolymer. Polyethyleneimine dissociates and carries a positive charge, while the adsorption ligand carries a negative charge. The positive charge of polyethyleneimine and the negative charge of the adsorption ligand are combined through electrostatic interaction, thus attaching the adsorption ligand to the polyethyleneimine. Therefore, the chloromethyl covalent grafting of polyethyleneimine generated during the reaction of the low-crosslinked styrene-divinylbenzene copolymer avoids the introduction of other reagents or substances. Furthermore, the covalent bonding of polyethyleneimine to the ultra-highly crosslinked styrene-divinylbenzene resin enhances the binding strength of polyethyleneimine on the resin, preventing polyethyleneimine detachment and ensuring the resin's adsorption performance. In addition, the electrostatic adsorption of the adsorption ligand to the polyethyleneimine facilitates its detachment from the protein-bound toxin adsorbent and its entry into the bloodstream during subsequent use, thus enabling the adsorption ligand to bind to the protein. The toxins compete for adsorption, causing protein-bound toxins to detach from albumin, increasing the free protein-bound toxins. On the other hand, the negatively charged adsorption ligands connect with polyethyleneimine through electrostatic interactions. When the adsorption ligands detach and enter the bloodstream, they bind to albumin under electrostatic interactions, reducing the number of free adsorption ligands in the blood. To maintain the stability of electrostatic interactions, the adsorption ligands are promoted to detach from the protein-bound toxins, so that electrostatic interactions and the detachment of adsorption ligands reach a balance. This further increases the free protein-bound toxins and enhances the adsorption capacity of the protein-bound toxin adsorbent for free protein-bound toxins.

[0038] In this embodiment, the loading of polyethyleneimine ranges from 0.1 mmol / g to 1 mmol / g, meaning that the loading of polyethyleneimine on each 1g of protein-binding toxin adsorbent is 0.1 mmol to 1 mmol. This allows for the electrostatic adsorption of an appropriate amount of adsorbent ligand, further improving the clearance rate of protein-binding toxins by the adsorbent. Simultaneously, it avoids excessive adsorbent ligand release into the bloodstream in a short time, which could cause discomfort and reduce the safety of the protein-binding toxin adsorbent.

[0039] It should be noted that, in this embodiment, the specific numerical ranges of the crosslinking degree of the ultra-high crosslinked styrene-divinylbenzene resin and the low crosslinked styrene-divinylbenzene copolymer are not further limited. Those skilled in the art can make adjustments according to the actual situation, as long as the crosslinking degree of the ultra-high crosslinked styrene-divinylbenzene resin is higher than that of the low crosslinked styrene-divinylbenzene copolymer.

[0040] Most protein-binding toxins contain aromatic rings and / or indole rings, and also possess ionic functional groups, making them readily bind to serum albumin and form large protein-bound morphologies. This results in a low free fraction of the toxin, and the distribution volume of the toxin in the body is much larger than the plasma volume, making it difficult to eliminate. Based on the above characteristics of protein-binding toxins, the inventors of this application have analyzed and provided a protein-binding toxin adsorbent. This adsorbent uses ultra-high cross-linked styrene-divinylbenzene resin as a carrier, immobilized with polyethyleneimine. The polyethyleneimine is connected to an adsorbent ligand, which carries a negative charge. When this adsorbent comes into contact with blood, the adsorbent ligand detaches from the adsorbent and enters the bloodstream. The adsorbent ligand has the same albumin-binding sites as p-cresol sulfate and indolephenol sulfate, and can compete with the protein-binding toxin for adsorption onto albumin. Furthermore, the binding force between the adsorbent ligand and albumin is stronger than the binding force between the protein-binding toxin and albumin. This process causes protein-bound toxins to detach from albumin, rendering them free and facilitating adsorption by the hypercrosslinked styrene-divinylbenzene resin and the polyethyleneimine immobilized on it, thus improving the clearance rate. Simultaneously, the hypercrosslinked styrene-divinylbenzene resin, with its abundant porous structure, can adsorb medium-to-large molecules and small-molecule protein-bound toxins in uremic toxins. The positively charged polyethyleneimine immobilized on the resin can electrostatically bind to free protein-bound toxins in the blood, thus clearing them. In this embodiment, the protein-bound toxins are rendered free through the action of the adsorption ligand, and simultaneously cleared through the adsorption of the hypercrosslinked styrene-divinylbenzene resin and the electrostatic binding of polyethyleneimine, thereby improving the clearance rate.

[0041] Figure 1 This is a process flow diagram for preparing protein-binding toxin adsorbents provided in the embodiments of this application. Figure 1 As shown, a second aspect of this application provides a method for preparing the protein-binding toxin adsorbent described in the first aspect, comprising the following steps:

[0042] Step S110: Prepare a low-crosslinked styrene-divinylbenzene copolymer.

[0043] Specifically, a low-crosslinked styrene-divinylbenzene copolymer is prepared by suspension polymerization of styrene monomers, polyvinyl crosslinking agents, pore-forming agents, and initiators in a dispersion medium.

[0044] More specifically, styrene monomers, polyvinyl crosslinking agents, porogens, and initiators are mixed to form an oil phase, while a dispersion medium, sodium chloride, and magnesium chloride are mixed to form an aqueous phase. The oil phase is added to the aqueous phase, and a suspension polymerization reaction is carried out at 50°C to 100°C with stirring. After reacting for 10 to 20 hours, the suspension polymerization product is washed clean with water and ethanol, dried, and a low-crosslinked styrene-divinylbenzene copolymer is obtained. During the suspension polymerization reaction, the temperature can be gradually increased in stages. For example, after the oil phase forms uniform droplets of a certain size in the aqueous phase, the temperature can be raised to 75°C for 5 hours, then raised to 80°C for 5 hours, then raised to 95°C to 98°C, and the reaction can continue for 6 to 8 hours before stopping the reaction.

[0045] The styrene monomer is selected from at least one of styrene, methylstyrene, and ethylstyrene; preferably, the styrene monomer is styrene. The polyvinyl crosslinking agent is selected from at least one of divinylbenzene, divinyltoluene, divinylxylene, and divinylethylbenzene; preferably, the polyvinyl crosslinking agent is divinylbenzene. The styrene monomer accounts for 20% to 90% of the total mass of the styrene monomer and the polyvinyl crosslinking agent, and the polyvinyl crosslinking agent accounts for 10% to 80% of the total mass of the styrene monomer and the polyvinyl crosslinking agent. Therefore, using monovinyl styrene monomers and polyvinyl crosslinking agents as reactive monomers allows for partial crosslinking of the styrene-divinylbenzene copolymer, which is beneficial for improving the mechanical strength and structural stability of the low-crosslinked styrene-divinylbenzene copolymer in organic solvents.

[0046] The porogen is a mixture of at least two substances selected from aromatics, alkanes, higher alcohols, higher ketones, and esters; wherein the aromatics are selected from toluene and xylene; the alkanes are selected from n-heptane, 200# gasoline, and solid paraffin; the higher alcohols are selected from butanol, hexanol, cyclohexanol, isooctanol, n-octanol, and methyl isobutyl methanol; the higher ketones are selected from methyl isobutyl ketone, 2-hexanone, diisobutyl ketone, and methyl tert-butyl ketone; and the esters are selected from butyl acetate, ethyl acetate, and butyl butyrate; preferably, the porogen is toluene and methyl isobutyl methanol. As an optional embodiment, the mass of the porogen can be 70% to 230% of the total mass of the styrene monomer and the polyvinyl crosslinking agent.

[0047] The initiator is selected from at least one of benzoyl peroxide, tert-butyl peroxide-2-ethylhexanoate, and tert-amyl peroxide-2-ethylhexanoate; preferably, the initiator is benzoyl peroxide. As an optional embodiment, the mass of the initiator is 0.5% to 1.5% of the total mass of the styrene monomer and the polyvinyl crosslinking agent. Therefore, the above-mentioned initiator can effectively initiate polymerization, and the initiator is inexpensive.

[0048] The dispersion medium is water, and the dispersion medium contains a dispersant selected from at least one of gelatin, polyvinyl alcohol, and carboxymethyl cellulose. Preferably, the dispersion medium is gelatin. As an optional embodiment, the mass of the dispersant is 0.5% to 2% of the mass of the dispersion medium. Thus, the above-mentioned dispersion medium is safe and environmentally friendly, and can achieve suspension polymerization of low-crosslinked styrene-divinylbenzene copolymers.

[0049] As an optional implementation, the mass of sodium chloride is 3% to 5% of the mass of the aqueous phase, and the mass of magnesium chloride is 2% to 4% of the mass of the aqueous phase. The mass of the aqueous phase is the sum of the masses of the dispersion medium, dispersant, sodium chloride, and magnesium chloride.

[0050] Step S120: The low-crosslinked styrene-divinylbenzene copolymer undergoes a crosslinking reaction to prepare a high-crosslinked styrene-divinylbenzene resin. Polyethyleneimine is immobilized on the high-crosslinked styrene-divinylbenzene resin during the crosslinking reaction of the low-crosslinked styrene-divinylbenzene copolymer.

[0051] Specifically, a low-crosslinked styrene-divinylbenzene copolymer is subjected to a chloromethylation reaction, and polyethyleneimine is added during the chloromethylation reaction to obtain a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine; the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine is subjected to a Friedel-Crafts reaction to obtain an ultra-high crosslinked styrene-divinylbenzene resin.

[0052] More specifically, the preparation of a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine includes the following steps:

[0053] The low-crosslinked styrene-divinylbenzene copolymer was added to dichloroethane and swollen at room temperature to ensure full swelling. The swollen low-crosslinked styrene-divinylbenzene copolymer, chloromethyl ether, and anhydrous ferric chloride were refluxed at 50°C to 60°C for 2 to 4 hours, and then the temperature was raised to 80°C to 100°C and refluxed for another 5 to 24 hours. Simultaneously, polyethyleneimine was added during the heating process to introduce polyethyleneimine into the low-crosslinked styrene-divinylbenzene copolymer, thus obtaining a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine. Therefore, after the low-crosslinked styrene-divinylbenzene copolymer and chloromethyl ether undergo a chloromethylation reaction at 50°C to 60°C, chloromethyl groups are introduced onto the low-crosslinked styrene-divinylbenzene copolymer. The temperature is then raised to 80°C to 100°C, and polyethyleneimine is added during the heating process. This allows the chloromethyl groups to react with the amino groups of polyethyleneimine, thereby introducing polyethyleneimine onto the low-crosslinked styrene-divinylbenzene copolymer. Simultaneously, the chloromethylation reaction continues during subsequent heating, introducing a sufficient amount of chloromethyl groups onto the low-crosslinked styrene-divinylbenzene copolymer, resulting in an excess of chloromethyl groups over polyethyleneimine. This embodiment... In the example, the chloromethyl covalently grafted polyethyleneimine generated during the reaction of the low-crosslinked styrene-divinylbenzene copolymer can, on the one hand, avoid the introduction of other reagents or substances, and the polyethyleneimine is covalently bonded to the ultra-high crosslinked styrene-divinylbenzene resin, which can improve the bonding strength of polyethyleneimine immobilized on the ultra-high crosslinked styrene-divinylbenzene resin, prevent polyethyleneimine from falling off and affecting the adsorption performance of the subsequently prepared ultra-high crosslinked styrene-divinylbenzene resin. On the other hand, it can ensure the introduction of sufficient chloromethyl groups, which is convenient for subsequent Friedel-Crafts reactions and improves the strength and structural stability of the support.

[0054] The mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to chloromethyl ether is 1:4 to 1:6; the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to anhydrous ferric chloride is 1:0.5 to 1:1.5; and the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to polyethyleneimine is 20:5 to 20:20, with the molecular weight of polyethyleneimine ranging from 200 to 10,000. By limiting the proportions of each substance within these ranges, sufficient chloromethyl groups are introduced into the low-crosslinked styrene-divinylbenzene copolymer, allowing some of the chloromethyl groups to react with polyethyleneimine, while the remaining chloromethyl groups undergo a subsequent Friedel-Crafts reaction to form a highly crosslinked styrene-divinylbenzene resin. In this embodiment, the chlorine content in the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine ranges from 1% to 5%.

[0055] After obtaining a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine, the copolymer was cleaned and then mixed with nitrobenzene. The mixture was allowed to swell at 35°C to 45°C for 4 to 5 hours. Zinc chloride was then added under stirring to induce a Friedel-Crafts reaction of the chloromethyl groups, forming a highly crosslinked styrene-divinylbenzene resin. The Friedel-Crafts reaction temperature was 110°C to 130°C, and the reaction time was 8 to 16 hours. This method allows the residual chloromethyl groups on the low-crosslinked styrene-divinylbenzene copolymer to undergo a Friedel-Crafts reaction, which improves the strength and structural stability of the resulting highly crosslinked styrene-divinylbenzene resin and facilitates the formation of more microporous channels, thereby enhancing the resin's adsorption capacity for small molecule protein-bound toxoids. In this embodiment, polyethyleneimine is immobilized on the ultra-high crosslinked styrene-divinylbenzene resin. The immobilization amount of polyethyleneimine ranges from 0.1 mmol / g to 1 mmol / g, that is, the immobilization amount of polyethyleneimine on each 1g protein-binding toxin adsorbent is 0.1 mmol to 1 mmol.

[0056] The mass of nitrobenzene is 5 to 7 times that of the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine; the mass of zinc chloride is 0.1 to 0.5 times that of the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

[0057] After obtaining the ultra-high crosslinked styrene-divinylbenzene resin, the process further includes: first washing the ultra-high crosslinked styrene-divinylbenzene resin with ethanol until it becomes clear, then washing it with 5% dilute hydrochloric acid, then extracting it with ethanol using a Soxhlet extraction method, and finally drying it to obtain purified ultra-high crosslinked styrene-divinylbenzene resin.

[0058] Step S130: Immerse the ultra-high cross-linked styrene-divinylbenzene resin in the adsorption ligand solution. After immersion, the adsorption ligand is linked to polyethyleneimine to obtain a protein-bound toxin adsorbent.

[0059] Specifically, ultra-highly cross-linked styrene-divinylbenzene resin is immersed in an alkaline solution until the pH value of the alkaline solution remains unchanged to obtain the transformed ultra-highly cross-linked styrene-divinylbenzene resin. After cleaning and drying, the transformed ultra-highly cross-linked styrene-divinylbenzene resin is immersed in an adsorption ligand solution. After immersion, it is cleaned to obtain a protein-binding toxin adsorbent. The mass percentage of adsorption ligand in the adsorption ligand solution is 10% to 40%, and the volume ratio of ultra-highly cross-linked styrene-divinylbenzene resin to adsorption ligand solution is 1:1.2 to 1:1.5. Therefore, by immersing the ultra-highly cross-linked styrene-divinylbenzene resin in an alkaline solution, the chloride ions on the ultra-highly cross-linked styrene-divinylbenzene resin are transformed into hydroxide ions. This facilitates the dissociation of polyethyleneimine on the ultra-highly cross-linked styrene-divinylbenzene resin, making the polyethyleneimine positively charged. This allows the positively charged polyethyleneimine to attract more negatively charged adsorbent ligands when the ultra-highly cross-linked styrene-divinylbenzene resin is immersed in the adsorbent ligand solution. Furthermore, the positive charge of the polyethyleneimine combines with the negatively charged adsorbent ligands through electrostatic interactions, thereby increasing the amount of adsorbent ligands attached to the polyethyleneimine. In addition, immersing the ultra-highly cross-linked styrene-divinylbenzene resin in the adsorbent ligand solution allows the adsorbent ligands to enter the pore structure of the ultra-highly cross-linked styrene-divinylbenzene resin, which is beneficial for the adsorbent ligands to fully contact and combine with the polyethyleneimine immobilized on the ultra-highly cross-linked styrene-divinylbenzene resin, thus increasing the amount of adsorbent ligands on the ultra-highly cross-linked styrene-divinylbenzene resin.

[0060] The alkaline solution is a NaOH solution with a concentration of 0.1 mol / L. The transformed hypercrosslinked styrene-divinylbenzene resin is immersed in the adsorbent ligand solution for 24 to 30 hours to ensure sufficient contact between the adsorbent ligand and the polyethyleneimine immobilized on the hypercrosslinked styrene-divinylbenzene resin. Preferably, when immersing the transformed hypercrosslinked styrene-divinylbenzene resin in the adsorbent ligand solution, the solution can be stirred. This method of stirring while immersing promotes the binding of the adsorbent ligand and the polyethyleneimine on the hypercrosslinked styrene-divinylbenzene resin.

[0061] In this embodiment, the transformed ultra-high crosslinked styrene-divinylbenzene resin is cleaned with deionized water, which avoids affecting the activity of the groups on the ultra-high crosslinked styrene-divinylbenzene resin and avoids changes in acidity and alkalinity that could affect the dissociation of polyethyleneimine on the ultra-high crosslinked styrene-divinylbenzene resin.

[0062] After the ultra-high crosslinked styrene-divinylbenzene resin is soaked in the adsorption ligand solution, it is washed with deionized water to obtain a protein-bound toxin adsorbent. Washing with deionized water can avoid affecting the binding between the adsorption ligand and polyethyleneimine, and prevent the adsorption ligand from falling off the polyethyleneimine, thus helping to ensure the structural stability of the protein-bound toxin adsorbent.

[0063] The method for preparing the protein-bound toxin adsorbent provided in this embodiment first prepares a low-crosslinked styrene-divinylbenzene copolymer through suspension polymerization, and then prepares a highly crosslinked ethylene-divinylbenzene resin from the low-crosslinked styrene-divinylbenzene copolymer. This highly crosslinked ethylene-divinylbenzene resin has a rich pore structure, which can adsorb medium and large molecular weight substances and small molecule protein-bound toxins in uremic toxins. Polyethyleneimine is added during the crosslinking reaction of the low-crosslinked styrene-divinylbenzene copolymer. The chloromethyl group generated during the reaction immobilizes the polyethyleneimine onto the highly crosslinked styrene-divinylbenzene resin, which avoids the introduction of other reagents or other substances. This not only simplifies the reaction process and improves the reaction efficiency, but also avoids affecting the adsorption performance of the highly crosslinked styrene-divinylbenzene resin. Subsequently, by immersing the highly crosslinked styrene-divinylbenzene resin in an adsorption ligand solution, the adsorption ligand can enter the pore structure of the highly crosslinked styrene-divinylbenzene resin, which is conducive to the full contact and binding of the adsorption ligand with the polyethyleneimine immobilized on the highly crosslinked styrene-divinylbenzene resin, and promotes the binding of polyethyleneimine with the adsorption ligand. The preparation method provided in this embodiment simplifies the reaction steps, makes the preparation process simple, the reaction conditions are mild, the safety during the reaction process is high, and the production cost is low, which is conducive to large-scale industrial production.

[0064] A third aspect of this application provides a hemoperfusion device comprising the protein-binding toxin adsorbent described above, or a protein-binding toxin adsorbent prepared by the method described above. Using the protein-binding toxin adsorbent provided in this application as the adsorbent material of the hemoperfusion device can effectively remove protein-binding toxins from the blood of uremia patients during hemoperfusion. Furthermore, this protein-binding toxin adsorbent is used externally and does not enter the body, thus posing minimal harm to the human body.

[0065] To provide a more detailed description of the present invention, specific embodiments will be used to further illustrate the invention. Unless otherwise specified, the experimental methods used in the embodiments of the present invention are conventional methods; unless otherwise specified, the materials and reagents used in the embodiments of the present invention are commercially available.

[0066] Example 1

[0067] This embodiment provides a method for preparing a protein-bound toxin adsorbent, comprising the following steps:

[0068] (1) Add 600 mL of an aqueous solution containing 2 wt% gelatin to a 1000 mL three-necked flask and stir at 50 °C for 2 h until the dispersant gelatin is completely dissolved. Then add 15 g of sodium chloride and 10 g of magnesium chloride and stir until completely dissolved to obtain an aqueous phase. Mix 42 g of styrene, 8 g of divinylbenzene (DVB), 50 g of toluene, 0.5 g of benzoyl peroxide, and 56 g of methyl isobutyl methanol (MIBC) to form an oil phase. Slowly add the oil phase to the aqueous phase and, under mechanical stirring, heat to 75 °C for 5 h, then heat to 80 °C for 5 h, then heat to 95 °C to 98 °C and hold for 6 h to 8 h to carry out a suspension polymerization reaction to crosslink and solidify the styrene-divinylbenzene copolymer and distill off the toluene. After the reaction is complete, wash the reaction product several times with water and ethanol, filter, dry, and sieve. Select resin with a particle size of 0.6 mm to 1.2 mm to obtain a low-crosslinked styrene-divinylbenzene copolymer.

[0069] (2) Add 20g of low crosslinked styrene-divinylbenzene copolymer to a 500mL three-necked flask, add 500mL of dichloroethane, and let stand at room temperature for 12h to fully swell; add 100g of chloromethyl ether and 10g of anhydrous ferric chloride to the swollen low crosslinked styrene-divinylbenzene copolymer in sequence, start the stirrer, heat to 50℃ and reflux for 3h, then heat to 80℃ and continue to react for 7h, then add 5g of polyethyleneimine, and continue to react at 80℃ for 14h. After the reaction is completed, cool to room temperature, filter out the mother liquor, extract with methanol for 12h, wash with water until there is no methanol odor, filter, and dry to obtain low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

[0070] (3) Take 20g of low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, add 8g of zinc chloride under mechanical stirring, heat at 120℃ for 8h, chloromethyl undergoes Friedel-Crafts reaction to form a super-high crosslinked network, after the reaction is completed, cool to room temperature, filter out the mother liquor, wash with ethanol until clear, wash with 5% dilute hydrochloric acid for 8h, extract with ethanol for 8h, and dry to obtain super-high crosslinked styrene-divinylbenzene resin, on which polyethyleneimine is immobilized.

[0071] (4) Take 20g of purified ultra-high crosslinked styrene-divinylbenzene resin and soak it in 100ml of 0.1mol / L NaOH solution. Replace the NaOH solution every 2h until the pH value of the solution remains unchanged to obtain the transformed ultra-high crosslinked styrene-divinylbenzene resin. Wash the transformed ultra-high crosslinked styrene-divinylbenzene resin thoroughly with deionized water, drain it, and then soak the transformed ultra-high crosslinked styrene-divinylbenzene resin in 20wt% ibuprofen solution for 24h. After soaking, wash it 5 times with deionized water to obtain the protein-bound toxin adsorbent.

[0072] Example 2

[0073] This embodiment provides a method for preparing a protein-bound toxin adsorbent, comprising the following steps:

[0074] (1) Add 600 mL of an aqueous solution containing 2 wt% gelatin to a 1000 mL three-necked flask and stir at 50 °C for 2 h until the dispersant gelatin is completely dissolved. Then add 15 g of sodium chloride and 10 g of magnesium chloride and stir until completely dissolved to obtain an aqueous phase. Mix 40 g of styrene, 10 g of divinylbenzene (DVB), 50 g of toluene, 0.5 g of benzoyl peroxide, and 56 g of methyl isobutyl methanol (MIBC) to form an oil phase. Slowly add the oil phase to the aqueous phase and, under mechanical stirring, heat to 75 °C for 5 h, then heat to 80 °C for 5 h, then heat to 95 °C to 98 °C and hold for 6 h to 8 h to carry out a suspension polymerization reaction to crosslink and solidify the styrene-divinylbenzene copolymer and distill off the toluene. After the reaction is complete, wash the reaction product several times with water and ethanol, filter, dry, and sieve. Select resin with a particle size of 0.6 mm to 1.2 mm to obtain a low-crosslinked styrene-divinylbenzene copolymer.

[0075] (2) Add 20g of low crosslinked styrene-divinylbenzene copolymer to a 500mL three-necked flask, add 500mL of dichloroethane, and let stand at room temperature for 12h to fully swell; add 100g of chloromethyl ether and 10g of anhydrous ferric chloride to the swollen low crosslinked styrene-divinylbenzene copolymer in sequence, start the stirrer, heat to 50℃ and reflux for 3h, then heat to 80℃ and continue to react for 7h, then add 10g of polyethyleneimine, and continue to react at 80℃ for 14h. After the reaction is completed, cool to room temperature, filter out the mother liquor, extract with methanol for 12h, wash with water until there is no methanol odor, filter, and dry to obtain low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

[0076] (3) Take 20g of low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, add 8g of zinc chloride under mechanical stirring, heat at 120℃ for 8h, chloromethyl undergoes Friedel-Crafts reaction to form a super-high crosslinked network, after the reaction is completed, cool to room temperature, filter out the mother liquor, wash with ethanol until clear, wash with 5% dilute hydrochloric acid for 8h, extract with ethanol for 8h, and dry to obtain super-high crosslinked styrene-divinylbenzene resin, on which polyethyleneimine is immobilized.

[0077] (4) Take 20g of purified ultra-high crosslinked styrene-divinylbenzene resin and soak it in 100ml of 0.1mol / L NaOH solution. Replace the NaOH solution every 2h until the pH value of the solution remains unchanged to obtain the transformed ultra-high crosslinked styrene-divinylbenzene resin. Wash the transformed ultra-high crosslinked styrene-divinylbenzene resin thoroughly with deionized water, drain it, and then soak the transformed ultra-high crosslinked styrene-divinylbenzene resin in 30wt% ibuprofen solution for 24h. After soaking, wash it 5 times with deionized water to obtain the protein-binding toxin adsorbent.

[0078] Comparative Example 1

[0079] The ultra-high crosslinked styrene-divinylbenzene resin loaded with polyethyleneimine prepared in step (3) of Example 1 was used as the adsorbent in Comparative Example 1.

[0080] Comparative Example 2

[0081] The adsorbent in the commercially available HA130 hemoperfusion device manufactured by Jianfan Biotechnology Group Co., Ltd. was used as the adsorbent in Comparative Example 2.

[0082] Adsorption performance test: Protein-bound toxin-plasma solutions with an initial concentration of IS of 2.5 mg / L, PCS of 2.5 mg / L, and IAA of 0.5 mg / L were prepared by external addition method. 1 mL of the protein-bound toxin adsorbent prepared in Examples 1 and 2, and 1 mL of the adsorbent from Comparative Examples 1 and 2 were added to 10 mL of each of the three protein-bound toxin-plasma solutions. The solutions were then shaken at 110 r / min for 2 h in a constant-temperature shaker at 37°C. Subsequently, the concentrations of different protein-bound toxins before and after adsorption were measured, and the adsorption rates of different adsorbents for different protein-bound toxins were calculated (for example, when calculating the adsorption rate for IS, 1 mL of the protein-bound toxin adsorbent prepared in Examples 1 and 2, and 1 mL of the adsorbent in Comparative Examples 1 and 2 were added to 10 mL of a protein-bound toxin-plasma solution containing an initial IS concentration of 2.5 mg / L, and the mixture was shaken at 110 r / min for 2 h in a constant temperature shaker at 37°C. The concentrations of IS before and after adsorption were then measured, and the adsorption rates of different adsorbents for IS were calculated). The concentrations of IS, PCS, and IAA were determined by HPLC analysis.

[0083] The adsorption rate is calculated using the formula: Cr(%) = (C0 - Ct) / C0 × 100%, where Cr is the rate of decrease in the concentration of the target protein-bound toxin, C0 is the detection concentration of the target protein-bound toxin before adsorption, and Ct is the detection concentration of the target protein-bound toxin after 2 hours of adsorption. The adsorption rates of the adsorbents for each protein-bound toxin in each embodiment, Comparative Example 1, and Comparative Example 2 are shown in Table 1.

[0084] Table 1

[0085]

[0086] As shown in Table 1, the protein-bound toxin adsorbents in Examples 1 and 2 exhibit significantly improved adsorption performance for IS, PCS, and IAA compared to the adsorbents in Comparative Examples 1 and 2. This indicates that the protein-bound toxin adsorbent provided in this invention has a superior removal effect on protein-bound toxins, especially for IS, PCS, and IAA. Furthermore, the protein-bound toxin adsorbent provided in this invention also possesses good safety performance and excellent mechanical strength, meeting the requirements for whole blood perfusion.

[0087] While the disclosure is as stated above, its scope of protection is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of this disclosure, and all such changes and modifications will fall within the protection scope of this invention.

Claims

1. A protein-bound toxin adsorbent, characterized in that, The protein-binding toxin adsorbent uses ultra-high cross-linked styrene-divinylbenzene resin as a carrier, on which polyethyleneimine is immobilized. The polyethyleneimine is connected to an adsorption ligand, which includes at least one of α-methyl-4-(2-methylpropyl)phenylacetic acid, oleic acid and linoleic acid. The ultra-high crosslinked styrene-divinylbenzene resin is prepared by crosslinking a low crosslinked styrene-divinylbenzene copolymer. The polyethyleneimine is introduced during the crosslinking reaction of the low crosslinked styrene-divinylbenzene copolymer. The adsorption ligand is attached to the polyethyleneimine by electrostatic adsorption.

2. The protein-binding toxin adsorbent according to claim 1, characterized in that, The loading of the polyethyleneimine ranges from 0.1 mmol / g to 1 mmol / g.

3. A method for preparing a protein-bound toxin adsorbent, characterized in that, The method for preparing the protein-binding toxin adsorbent as described in claim 1 or 2 comprises: Preparation of low-crosslinked styrene-divinylbenzene copolymer; The low-crosslinked styrene-divinylbenzene copolymer undergoes a crosslinking reaction to prepare a highly crosslinked styrene-divinylbenzene resin, wherein polyethyleneimine is immobilized on the highly crosslinked styrene-divinylbenzene resin, and the polyethyleneimine is immobilized onto the highly crosslinked styrene-divinylbenzene resin during the crosslinking reaction of the low-crosslinked styrene-divinylbenzene copolymer. The ultra-high cross-linked styrene-divinylbenzene resin was immersed in an adsorption ligand solution. After immersion, the adsorption ligand was attached to the polyethyleneimine to obtain a protein-bound toxin adsorbent.

4. The method for preparing the protein-binding toxin adsorbent according to claim 3, characterized in that, The low-crosslinked styrene-divinylbenzene copolymer undergoes a crosslinking reaction to prepare a highly crosslinked styrene-divinylbenzene resin, comprising: The low crosslinked styrene-divinylbenzene copolymer was subjected to a chloromethylation reaction, and polyethyleneimine was added during the chloromethylation reaction to obtain a low crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine. The low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine was subjected to a Friedel-Crafts reaction to obtain a highly crosslinked styrene-divinylbenzene resin.

5. The method for preparing the protein-binding toxin adsorbent according to claim 4, characterized in that, The step of subjecting the low-crosslinked styrene-divinylbenzene copolymer to a chloromethylation reaction, wherein polyethyleneimine is added during the chloromethylation reaction, to obtain a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine, comprises: After swelling in dichloroethane, the low-crosslinked styrene-divinylbenzene copolymer, chloromethyl ether, and anhydrous ferric chloride are refluxed at 50°C to 60°C for 2 to 4 hours. Then, the temperature is raised to 80°C to 100°C and refluxed for another 5 to 24 hours. Simultaneously, polyethyleneimine is added during the heating process to introduce the polyethyleneimine onto the low-crosslinked styrene-divinylbenzene copolymer, thus obtaining a low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine.

6. The method for preparing the protein-binding toxin adsorbent according to claim 5, characterized in that, The mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the chloromethyl ether is 1:4 to 1:6; the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the anhydrous ferric chloride is 1:0.5 to 1:1.5; and the mass ratio of the low-crosslinked styrene-divinylbenzene copolymer to the polyethyleneimine is 20:5 to 20:

20.

7. The method for preparing the protein-binding toxin adsorbent according to claim 4, characterized in that, The process of subjecting the low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine to a Friedel-Crafts reaction to obtain a highly crosslinked styrene-divinylbenzene resin comprises: The low-crosslinked styrene-divinylbenzene copolymer containing polyethyleneimine is mixed with nitrobenzene and allowed to stand at 35°C to 45°C for 4 to 5 hours. Zinc chloride is then added under stirring to carry out a Friedel-Crafts reaction to form a highly crosslinked styrene-divinylbenzene resin. The Friedel-Crafts reaction is carried out at a temperature of 110°C to 130°C for 8 to 16 hours.

8. The method for preparing the protein-binding toxin adsorbent according to claim 3, characterized in that, The process involves immersing the ultra-highly cross-linked styrene-divinylbenzene resin in an adsorption ligand solution. After immersion, the adsorption ligand is attached to the polyethyleneimine to prepare a protein-binding toxin adsorbent, comprising: The ultra-highly crosslinked styrene-divinylbenzene resin is immersed in an alkaline solution until the pH value of the alkaline solution remains unchanged to obtain the transformed ultra-highly crosslinked styrene-divinylbenzene resin. After the transformed ultra-highly crosslinked styrene-divinylbenzene resin is cleaned and dried, it is immersed in an adsorbent ligand solution. After immersion, it is cleaned to obtain a protein-binding toxin adsorbent, wherein the mass percentage of the adsorbent ligand in the adsorbent ligand solution is 10% to 40%.

9. The method for preparing the protein-bound toxin adsorbent according to claim 8, characterized in that, The transformed ultra-high crosslinked styrene-divinylbenzene resin was cleaned with deionized water after being soaked in the adsorbent ligand.

10. A blood perfusion device, characterized in that, Includes the protein-binding toxin adsorbent as described in claim 1 or 2, or the protein-binding toxin adsorbent prepared by the preparation method described in any one of claims 3 to 9.

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