Blood purification device

Abstract

The invention provides a blood purification device, which comprises a blood perfusion device, the blood perfusion device is filled with adsorption resin, and the adsorption resin is prepared by the following steps: mixing a reaction monomer and an epoxy group-containing compound, and directly carrying out suspension polymerization reaction on the epoxy group-containing compound and the reaction monomer through carbon-carbon double bonds to obtain the adsorption resin. The macroporous adsorption resin containing the epoxy group is prepared; the method comprises the following steps: preparing a cationic ligand solution from a cationic ligand, carrying out a first-stage reaction on macroporous adsorption resin containing epoxy groups and the cationic ligand solution at a first preset temperature under an alkaline condition for a first time, heating to a second preset temperature, carrying out a second-stage reaction for a second time, and carrying out a second-stage reaction for a second time, and reacting amido in the cationic ligand with the epoxy group, and immobilizing the reaction product onto macroporous adsorption resin containing the epoxy group to prepare the adsorption resin. The blood purification device can adsorb endotoxin and cell factors at the same time.

Description

Technical Field

[0001] The present invention relates to the technical field of blood purification, and in particular to a blood purification device. Background Art

[0002] Sepsis typically begins with infection by bacteria, viruses, fungi, or other microorganisms. When these pathogens invade the body and begin to multiply, they release endotoxins, which stimulate and activate the patient's immune system, leading to the production and release of excessive cytokines, including tumor necrosis factor α (TNF-α) and various interleukins (such as IL-1 and IL-6), a phenomenon known as a "cytokine storm." Cytokines trigger a systemic inflammatory response, increase vascular permeability, and cause plasma to efflux into the interstitial space, leading to edema and reduced circulating blood volume. This damages endothelial cells, activates coagulation mechanisms, and forms microthrombi, further impairing organ function. Both endotoxins and cytokines play a key role in the development and progression of sepsis. In addition to sepsis, the pathogenic mechanisms of endotoxemia, septic shock, multiple organ dysfunction syndrome (MODS), severe trauma, liver disease, and acute pancreatitis are also closely related to endotoxins and cytokines. These critical illnesses require timely and effective medical treatment to save the patient's life. Therefore, the simultaneous elimination of excess endotoxins and cytokines in the patient's body is of great clinical significance.

[0003] Currently, there are no medical devices, drugs, or other products in clinical practice that can simultaneously adsorb endotoxins and cytokines. Existing adsorbent products generally adsorb only endotoxins or only cytokines. For example, Toray (Japan), Alteco (Sweden), Pocard (Russia), and Efferon (Russia) have developed products that adsorb only endotoxins, while CytoSorbents (USA) has developed products that adsorb only cytokines. Summary of the Invention

[0004] The present invention aims to provide a blood purification device, wherein the adsorption resin in the blood purification device can simultaneously adsorb endotoxins and cytokines, and the residual substances in the adsorption resin are relatively small.

[0005] To solve the above problems, the present invention provides a blood purification device, including a blood perfusion device, wherein the blood perfusion device is filled with an adsorption resin, and the adsorption resin is prepared by the following method:

[0006] Mixing a reactive monomer and an epoxy-containing compound, wherein the reactive monomer is a mixture of a styrene monomer and a polyvinyl crosslinking agent, or the reactive monomer is a polyvinyl crosslinking agent, and the epoxy-containing compound has a carbon-carbon double bond, and the epoxy-containing compound directly reacts with the reactive monomer via the carbon-carbon double bond to undergo a suspension polymerization reaction to prepare a macroporous adsorption resin containing an epoxy group;

[0007] The cationic ligand is prepared into a cationic ligand solution, wherein the cationic ligand is a monomer having a hydrophobic fatty chain at one end and an amine group at at least one end;

[0008] Under alkaline conditions, the macroporous adsorption resin containing epoxy groups and the cationic ligand solution are subjected to a first-stage reaction at a first preset temperature. After the first reaction period, the temperature is raised to a second preset temperature for a second-stage reaction. The reaction period is second, so that the amino groups in the cationic ligand react with the epoxy groups and are immobilized on the macroporous adsorption resin containing epoxy groups to obtain the adsorption resin.

[0009] The blood purification device described in the present invention first directly carries out a suspension polymerization reaction between the reactive monomer and the epoxy-containing compound, so that the epoxy groups are embedded in the skeleton structure of the macroporous adsorption resin. Compared with the scheme of grafting epoxy groups on the macroporous adsorption resin, the pore structure and skeleton are less affected. Not only can the influence of the epoxy groups on the pore structure of the macroporous adsorption resin be reduced, but the hydrophobic characteristics of the skeleton can be maintained by controlling the amount of epoxy groups grafted, so that the macroporous adsorption resin can have a good adsorption effect on various cytokines and inflammatory mediators represented by IL-6 in the blood through molecular sieving and lipophilic-hydrophobic effects. In addition, the reactive monomer and the epoxy-containing compound are directly subjected to a suspension polymerization reaction, without utilizing the residual double bonds of the macroporous adsorption resin to graft the epoxy-containing compound, so that the epoxy group content on the macroporous adsorption resin can be controlled, and the fixed loading amount of epoxy groups on the macroporous adsorption resin can be accurately controlled, so that cationic ligands can be selected and grafted to achieve good adsorption performance for both cytokines and endotoxins. Subsequently, the cationic ligand is formulated into a cationic ligand solution, which can enable the cationic ligand and the macroporous adsorption resin containing epoxy groups to fully react, and the cationic ligand and the macroporous adsorption resin containing epoxy groups first undergo a first-stage reaction at a first preset temperature, and then the temperature is raised to a second preset temperature for a second-stage reaction. Through the staged temperature increase reaction, the amino group in the cationic ligand can gradually react with the epoxy group, so that the cationic ligand is fixed on the macroporous adsorption resin, which not only improves the connection tightness between the cationic ligand and the macroporous adsorption resin, but also improves the grafting rate of the cationic ligand. After the cationic ligand is grafted onto the skeleton of the macroporous adsorption resin, a secondary amine structure is formed, which has a strong electrostatic adsorption effect in the solution and can form a strong electrostatic force with the phosphate group on the endotoxin lipid A, and adsorb endotoxin through electrostatic action. At the same time The hydrophobic fatty chains in the cationic ligand can be adsorbed together with the long fatty chains of the lipid A part of the endotoxin through hydrophobic interaction. Through the form of dual adsorption sites of electrostatic adsorption and hydrophobic adsorption, the endotoxin can be captured or anchored, thereby improving the adsorption capacity of the adsorption resin used for blood perfusion for endotoxin; in addition, after the macroporous adsorption resin is grafted with the cationic ligand, the active hydrophobic fatty chains are distributed on the outside of the macroporous adsorption resin and the inner surface of the pores, which can improve the hydrophobic properties of the macroporous adsorption resin. The hydrophobic fatty chains on the cationic ligand can move lipophilic and hydrophobic macromolecules such as cytokines toward the macroporous adsorption resin through lipophilic and hydrophobic interactions, and then through the combination of the hydrophobic fatty chains on the cationic ligand and the hydrophobic interaction of the macroporous adsorption resin itself, the adsorption of lipophilic and hydrophobic macromolecules such as cytokines and endotoxins containing hydrophobic lipid A can be greatly improved.

[0010] Compared with the existing preparation method of modifying and grafting on styrene resin microspheres, the method for preparing the adsorption resin of the present invention is simple and efficient, has mild reaction conditions, is suitable for industrial production, and only requires two steps to complete the preparation of a cross-linked adsorption resin. There is no need to modify the adsorption resin through a multi-step intermediate grafting process, nor is there any need for further cross-linking to further improve the performance of the adsorption resin. It avoids the introduction of new initiators or catalysts and other substances that lead to excessive residues or excessive side reaction products. This preparation method is specifically designed according to the molecular structural characteristics of endotoxin, greatly improving the adsorption capacity for endotoxin, so that the prepared adsorption resin has a good adsorption effect on both endotoxin and cytokines. The adsorption resin used for blood perfusion has an adsorption rate of IL-6 and endotoxin of more than 85%, an adsorption rate of TNF-α of more than 58%, and an adsorption rate of β2 microglobulin of up to 95%. Compared with the existing adsorption resin with an adsorption rate of IL-6 below 75%, an adsorption rate of TNF-α below 45%, an adsorption rate of β2 microglobulin below 74%, and an adsorption rate of endotoxin below 64%, the adsorption performance of the adsorption resin prepared by this preparation method for endotoxin and cytokines can be effectively improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 A process flow chart for preparing an adsorption resin for hemoperfusion provided in an embodiment of the present invention;

[0012] Figure 2 This is a schematic structural diagram of a hemoperfusion device provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0013] Currently, there are no medical devices, drugs, or other products that can adsorb endotoxins and cytokines simultaneously in the clinic. Existing adsorbent products can basically only adsorb endotoxins or only adsorb cytokines. For example, Japan's Toray, Sweden's Alteco, Russia's Pocard, and Efferon have developed products that can only adsorb endotoxins, and the United States' CytoSorbents has developed products that can only adsorb cytokines. At the same time, although the existing technology provides many adsorbent preparation schemes, the adsorbents prepared by the existing technology either have a single function and cannot adsorb endotoxins and cytokines at the same time; or the adsorbents prepared by the existing technology can adsorb both substances at the same time, but there are problems such as complex preparation methods, a large amount of residual substances, and the safety and effectiveness cannot be guaranteed or the cost is high. In addition, the adsorbents prepared by the existing technology that can adsorb both substances at the same time mainly adsorb one of the two substances, and the other substance is only adsorbed incidentally, and can only be adsorbed in small amounts, with weak adsorption capacity.

[0014] In order to solve the above problems existing in the prior art, this embodiment provides an adsorption resin having good adsorption properties for both cytokines and endotoxins. The adsorption resin can be used in a blood purification device and filled in a blood perfusion device.

[0015] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0016] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0017] In addition, the terms "comprising", "including", "containing", and "having" are not restrictive, and other steps and other components that do not affect the results can be added. Unless otherwise specified, materials, equipment, and reagents are commercially available.

[0018] In addition, although the present invention describes the steps in the preparation in the form of S110 and S120, this description is only for ease of understanding, and the form of S110 and S120 does not limit the sequence of the steps.

[0019] Figure 1 The process flow chart of preparing adsorption resin for blood perfusion provided in the examples of this application. Figure 1 As shown, a first aspect of the embodiments of the present application provides a method for preparing an adsorption resin for blood perfusion, comprising:

[0020] Step S110, subjecting the reaction monomer and the epoxy-containing compound to a suspension polymerization reaction to obtain a macroporous adsorption resin containing epoxy groups; wherein the reaction monomer is a mixture of a styrene monomer and a polyvinyl crosslinking agent, or the reaction monomer is a polyvinyl crosslinking agent; and the epoxy-containing compound has a carbon-carbon double bond.

[0021] Specifically, the reaction monomer and the epoxy-containing compound are subjected to a suspension polymerization reaction in a dispersion medium under the action of a porogen and an initiator. After the suspension polymerization reaction is completed, a macroporous adsorption resin containing epoxy groups is obtained; wherein the suspension polymerization reaction temperature is 70°C to 90°C, and the suspension polymerization reaction time is 4h to 12h.

[0022] In this embodiment, by subjecting the reactive monomer and the epoxy-containing compound to a suspension polymerization reaction in a dispersion medium under the action of a porogen and an initiator, the epoxy-containing compound can directly undergo a suspension polymerization reaction with the reactive monomer through a carbon-carbon double bond, so that the epoxy-containing compound can be directly polymerized onto a macroporous adsorption resin. This method is used to prepare a macroporous adsorption resin containing epoxy groups, and the epoxy groups can be embedded in the skeleton structure of the macroporous adsorption resin through a suspension polymerization reaction. The epoxy groups have little effect on the pore structure of the macroporous adsorption resin, so that the macroporous adsorption resin still has the characteristics of a porous structure and a hydrophobic skeleton. The macroporous adsorption resin has better adsorption performance through molecular sieving and lipophilic-hydrophobic effects, which is beneficial to reducing the levels of various cytokines and inflammatory mediators represented by IL-6 in the blood. In addition, this method does not require the use of residual double bonds of the macroporous adsorption resin to graft epoxy-containing compounds. It not only improves the grafting rate of epoxy-containing compounds, but also realizes the controllability of the epoxy group content on the macroporous adsorption resin. It can accurately control the immobilization amount of epoxy groups on the macroporous adsorption resin, so that cationic ligands can be grafted in a targeted manner according to the structural characteristics of cytokines and endotoxins to achieve better adsorption performance for both cytokines and endotoxins.

[0023] Based on the above example, as an optional embodiment, the epoxy group content of the epoxy-containing macroporous adsorption resin is greater than 5 μmol / mL. That is, the immobilized epoxy group content per mL of the epoxy-containing macroporous adsorption resin is greater than 5 μmol. Having an epoxy group content within this range on the epoxy-containing macroporous adsorption resin facilitates the grafting of an appropriate amount of cationic ligands, thereby enhancing the simultaneous adsorption of cytokines and endotoxins.

[0024] Based on the above embodiment, as an optional embodiment, the styrene monomer is a mixture of one or more of styrene, methyl styrene, and ethyl styrene. If the styrene monomer is a mixture of the above, these substances can be mixed in any proportion. Of course, the styrene monomer can also be other compounds containing a single vinyl group, and those skilled in the art can make the selection based on actual conditions. The polyvinyl crosslinker is a mixture of one or more of divinylbenzene (DVB), divinyltoluene, divinylxylene, vinylethylbenzene, and ethylene glycol dimethacrylate (EGDMA). If the polyvinyl crosslinker is a mixture of the above, these substances can be mixed in any proportion. Of course, the polyvinyl crosslinker can also be a combination of one or more of other olefin organic compounds containing two or more carbon-carbon double bonds, and those skilled in the art can make the selection based on actual conditions. The amount of styrene monomer used is 0% to 50% of the mass of the reaction monomer, that is, the reaction monomer may not contain styrene monomer and only contain polyvinyl crosslinker. If the reaction monomer contains styrene monomer, the amount of styrene monomer used does not exceed 50% of the total mass of the styrene monomer and polyvinyl crosslinker. Preferably, the amount of styrene monomer used is 0% to 20% by mass of the reaction monomer. Thus, by controlling the amount of styrene monomer in the reaction monomer within the above range, on the one hand, it is beneficial to adjust the crosslinking degree of the macroporous adsorption resin containing epoxy groups, so that the crosslinking degree of the macroporous adsorption resin containing epoxy groups is within an appropriate range, which can ensure that the macroporous adsorption resin containing epoxy groups forms balls and avoid cracking caused by excessive rigidity. On the other hand, it is beneficial for the carbon-carbon double bond of the epoxy-containing compound to directly react with the reaction monomer, so that the epoxy-containing compound can be directly polymerized on the macroporous adsorption resin, thereby improving the stability of the epoxy group immobilization.

[0025] Based on the above embodiment, as an optional embodiment, the epoxy-containing compound contains both epoxy groups and carbon-carbon double bonds, and may include a mixture of one or both of glycidyl methacrylate and allyl glycidyl ether. If the epoxy-containing compound is a mixture of the above two substances, the two substances can be mixed in any proportion. Of course, the epoxy-containing compound can also be other compounds containing epoxy groups and carbon-carbon double bonds, wherein the number of epoxy groups and carbon-carbon double bonds can be two or more. The amount of the epoxy-containing compound used is 0.5% to 50% of the mass of the reaction monomers, preferably 1% to 15% of the mass of the reaction monomers. Thus, the epoxy-containing compound containing both epoxy groups and carbon-carbon double bonds can be directly polymerized onto the macroporous adsorption resin and subsequently react with the cationic ligand, thereby immobilizing the cationic ligand on the macroporous adsorption resin. By controlling the amount of the epoxy-containing compound within the above range, the content of epoxy groups immobilized on the macroporous adsorption resin can be precisely controlled, so that the content of epoxy groups on the macroporous adsorption resin is greater than 5 μmol / mL.

[0026] Based on the above embodiment, as an optional embodiment, the porogen is a mixture of two or more of aromatic hydrocarbons, alkanes, higher alcohols, and higher ketones, and the porogen includes a good solvent and a poor solvent. Aromatic hydrocarbons and alkanes are good solvents, with aromatic hydrocarbons including at least one of toluene, ethylbenzene, xylene, and n-propylbenzene, and alkanes including at least one of n-heptane, liquid paraffin, and gasoline. Higher alcohols and higher ketones are poor solvents, with higher alcohols including at least one of butanol, isooctyl alcohol, and methyl isobutyl carbinol, and higher ketones including at least one of methyl isobutyl ketone and 2-hexanone. The amount of the poor solvent used is 30% to 200% of the mass of the good solvent, preferably 50% to 100% of the mass of the good solvent. Thus, through the combination of good and poor solvents, the pore structure and morphology of the macroporous adsorption resin can be adjusted, ensuring that the macroporous adsorption resin has a good adsorption effect on endotoxins and cytokines. Controlling the amount of the poor solvent within the above range can ensure that the macroporous adsorption resin has a better adsorption effect on macromolecular substances such as cytokines and endotoxins. In this embodiment, the amount of the porogen used is 10% to 300% of the total mass of the reaction monomers and the epoxy-containing compound. Preferably, the amount of the porogen used is 100% to 200% of the total mass of the reaction monomers and the epoxy-containing compound, which is beneficial for adjusting the pore structure of the macroporous adsorption resin containing epoxy groups, so that its pore volume and pore diameter are within an appropriate range.

[0027] Based on the above embodiment, as an alternative embodiment, the initiators are organic peroxides and alkylphenyl sulfones. The organic peroxides are selected from one or more of benzoyl peroxide, tert-butyl peroxy-2-hexylhexanoate, tert-amyl peroxy-2-ethylhexanoate, and dodecyldibenzoyl peroxide; and the alkylphenyl sulfones are selected from one or more of dihydroxyphenyl sulfone and tert-butyl phenyl sulfone. If the initiator is a mixture of several substances, the substances can be mixed in any proportion. Preferably, the initiator is tert-butyl peroxy-2-hexylhexanoate. The amount of the initiator is 0.1% to 10% of the total mass of the reaction monomers and the epoxy-containing compound, and more preferably, the amount of the initiator is 0.5% to 5% of the total mass of the reaction monomers and the epoxy-containing compound. Therefore, using the above-mentioned substances as initiators and limiting the initiator amount to the above-mentioned range facilitates initiation of the suspension polymerization reaction of the reaction monomers and the epoxy-containing compound.

[0028] Based on the above embodiment, as an alternative embodiment, the dispersion medium includes a dispersant and water, wherein the dispersant is dissolved in the water to form the dispersion medium. The dispersant comprises one or a mixture of gelatin, polyvinyl alcohol, and methylcellulose, and the amount of the dispersant is 0.01% to 5% by mass of the dispersion medium. If the dispersant is a mixture of several substances, the substances can be mixed in any proportion. Therefore, selecting the above-mentioned substances as the dispersant and limiting the amount of the dispersant to within the above-mentioned range facilitates adjusting the particle size of the epoxy-containing macroporous adsorption resin.

[0029] In this embodiment, the volume ratio of the dispersion medium to the mixed organic phase is 1:2 to 5:1, where the mixed organic phase refers to a mixture of the reactive monomers comprising an epoxy compound, a porogen, and an initiator. By controlling the volume ratio of the dispersion medium to the mixed organic phase, the stability of the suspension polymerization reaction is ensured, and the resin exhibits a smooth, spherical shape after being formed.

[0030] The macroporous adsorption resin containing epoxy groups prepared by the method of this embodiment has a particle size range of 0.15 mm to 1.5 mm and a specific surface area (measured by BET method) range of 250 m 2 / g to 1500m 2 / g, and the average pore size ranges from 5nm to 150nm. Thus, it can be ensured that the subsequently prepared macroporous adsorption resin containing epoxy groups has a good adsorption capacity for cytokines in the blood.

[0031] Step S120: Under alkaline conditions, grafting cationic ligands onto a macroporous adsorption resin containing epoxy groups to prepare an adsorption resin for blood perfusion; wherein the cationic ligand is a monomer containing a hydrophobic fatty chain at one end and an amino group at at least one end.

[0032] Specifically, the cationic ligand is prepared into a cationic ligand solution; under alkaline conditions, the macroporous adsorption resin containing epoxy groups and the cationic ligand solution are subjected to a first-stage reaction at a first preset temperature. After the first reaction period, the temperature is raised to a second preset temperature for a second-stage reaction. After the second reaction period, an adsorption resin for blood perfusion is obtained.

[0033] In this embodiment, by first preparing the cationic ligand into a cationic ligand solution, it is possible to ensure that the cationic ligand and the macroporous adsorption resin containing epoxy groups fully react. The two first undergo a first-stage reaction at a first preset temperature. The first preset temperature is relatively low, which can avoid excessive loss due to the ring opening of all epoxy groups and affect the grafting rate. The temperature is then raised to a second preset temperature for a second-stage reaction. The temperature increase allows the remaining epoxy groups to be further ring-opened and grafted, allowing the remaining epoxy groups to be grafted with more cationic ligands, while also avoiding residual epoxy groups. In this embodiment, by conducting the reaction in stages, the grafting rate of the cationic ligand can be increased and residual epoxy groups can be avoided. In this embodiment, cationic ligands are grafted onto a macroporous adsorption resin containing epoxy groups, and the amino groups in the cationic ligands react with the epoxy groups to be immobilized on the macroporous adsorption resin, thereby improving the stability of the cationic ligands grafted onto the macroporous adsorption resin. The cationic ligands contain amino groups and hydrophobic fatty chains, which enable them to capture and anchor endotoxins under the dual effects of electrostatic adsorption and lipophilic and hydrophobic properties, thereby improving the adsorption performance of endotoxins. In addition, the hydrophobic fatty chains on the cationic ligands can also improve the hydrophobic properties of the macroporous adsorption resin, so that the macroporous adsorption resin will not be too hydrophilic. Therefore, the hydrophobic fatty chains on the cationic ligands are combined with the hydrophobic effect of the macroporous adsorption resin itself to achieve a significant improvement in the adsorption of lipophilic and hydrophobic macromolecules such as cytokines.

[0034] Based on the above embodiments, as an optional implementation, the cationic ligand can be a monomer containing an amine group (primary amine) at one end and a hydrophobic fatty chain at the other end. For example, monomers such as butylamine, hexylamine, and hexadecylamine can be selected; the cationic ligand can also be a monomer containing an amine group (primary amine) at both ends and a hydrophobic fatty chain at the middle end. For example, monomers such as octanediamine, decanediamine, and lysine can be selected. By selecting a cationic ligand containing both an amine group and a hydrophobic fatty chain, a strong adsorption effect can be produced on the lipid A portion of the endotoxin. Since lipid A contains a negatively charged phosphate group, after the cationic ligand is grafted onto the macroporous adsorption resin skeleton, a secondary amine structure is formed, which has a strong electrostatic adsorption effect in the solution and can form a strong electrostatic force with the phosphate group on the endotoxin lipid A, thereby adsorbing endotoxin through electrostatic action. At the same time, lipid A also contains a hydrophobic fatty long chain. The hydrophobic fatty chain in the cationic ligand can be adsorbed together with the fatty long chain of the endotoxin lipid A portion through hydrophobic action, and the endotoxin is adsorbed through the hydrophobic force, realizing the form of dual adsorption sites of electrostatic adsorption and hydrophobic adsorption, which can improve the adsorption capacity of the adsorption resin used for blood perfusion for endotoxin.

[0035] Based on the above embodiment, as an optional embodiment, the hydrophobic fatty chain in the cationic ligand contains 4 to 30 carbon atoms. This can avoid the situation where the number of carbon atoms in the hydrophobic fatty chain is too small, which makes the hydrophobic adsorption force of the cationic ligand weaker and affects the adsorption performance of endotoxins. It can also avoid the situation where the number of carbon atoms in the hydrophobic fatty chain is too large, which leads to diffusion obstruction during the grafting of the cationic ligand onto the macroporous adsorption resin due to the excessive molecular weight of the cationic ligand, resulting in a low grafting rate. At the same time, the excessive molecular weight of the grafted cationic ligand may change the structure of the macroporous adsorption resin, adversely affecting the adsorption of cytokines. Therefore, limiting the number of carbon atoms in the hydrophobic fatty chain in the cationic ligand to the above range can ensure that the final adsorption resin has a good adsorption effect on both cytokines and endotoxins. Preferably, the hydrophobic fatty chain in the cationic ligand contains 5 to 16 carbon atoms.

[0036] Based on the above embodiment, as an optional implementation, the first preset temperature is 25°C to 60°C, the first duration is 1 to 24 hours, and the second preset temperature is 60°C to 85°C, the second duration is 1 to 24 hours. Setting the first and second preset temperatures within these ranges further improves the grafting rate of the cationic ligand and avoids residual epoxy groups.

[0037] Based on the above embodiment, as an optional embodiment, the volume ratio of the cationic ligand solution to the epoxy-containing macroporous adsorption resin is 1:3 to 5:1, and the mass fraction of the cationic ligand in the cationic ligand solution is 1% to 100%. This ensures that the cationic ligand solution and the epoxy-containing macroporous adsorption resin fully react. In this embodiment, the cationic ligand can be dissolved in water or an organic solvent to prepare the cationic ligand solution. If water is used to dissolve the cationic ligand, an alkaline substance such as sodium hydroxide or potassium hydroxide can be used to adjust the pH of the cationic ligand solution to accelerate the ring-opening reaction and achieve complete reaction. If an organic solvent is used to dissolve the cationic ligand, the organic solvent can be an ethanol solution, and the concentration of the ethanol solution can be greater than 0 and less than 100%.

[0038] Based on the above embodiment, as an optional implementation, in step S120, after the first stage reaction for the first time period and before the temperature is raised to the second preset temperature for the second stage reaction, the following steps are further included:

[0039] An amino acid compound is added to the reaction product of the first stage and mixed evenly to obtain a grafting mixture; wherein the amino acid compound is a neutral amino acid compound or a basic amino acid compound.

[0040] In this embodiment, by adding an amino acid compound to the first-stage reaction product, the biocompatibility of the adsorption resin used for blood perfusion is improved. The amino acid compound is a neutral amino acid compound or a basic amino acid compound, which can avoid the acidic amino acids (aspartic acid or glutamic acid) causing the electrostatic adsorption force of the adsorption resin used for blood perfusion to be weakened, thereby affecting the adsorption capacity of endotoxins.

[0041] Based on the above example, as an alternative embodiment, the mass fraction of the amino acid compound in the grafting mixture does not exceed 5%. This prevents excessive addition of the amino acid compound, which could alter the hydrophobic properties of the adsorption resin used for hemoperfusion, affecting the lipophilic and hydrophobic interactions of the adsorption resin with respect to large molecular weight endotoxins and cytokines, and thus affecting the adsorption capacity for endotoxins and cytokines.

[0042] Based on the above embodiment, as an optional embodiment, the amino acid compound is a mixture of one or more of serine, threonine, arginine, and histidine. If the amino acid compound is a mixture of several substances, these substances can be mixed in any proportion. Of course, those skilled in the art can also select other neutral amino acid compounds or basic amino acid compounds according to actual conditions.

[0043] The method for preparing an adsorbent resin for blood perfusion provided in this embodiment first directly conducts a suspension polymerization reaction between a reactive monomer and an epoxy-containing compound, so that the epoxy groups can be embedded in the backbone structure of the macroporous adsorbent resin. Compared with grafting epoxy groups onto the macroporous adsorbent resin, this method not only reduces the impact of the epoxy groups on the pore structure of the macroporous adsorbent resin, but also allows the macroporous adsorbent resin to retain the characteristics of a porous structure and a hydrophobic backbone. The macroporous adsorbent resin can effectively adsorb various cytokines and inflammatory mediators represented by IL-6 in the blood through molecular sieving and lipophilic-hydrophobic interactions. In addition, the direct suspension polymerization reaction between the reactive monomer and the epoxy-containing compound eliminates the need to utilize the residual double bonds of the macroporous adsorbent resin to graft the epoxy-containing compound. This allows for controllable epoxy group content on the macroporous adsorbent resin and precise control of the epoxy group loading on the macroporous adsorbent resin, thereby enabling targeted selection and grafting of cationic ligands to achieve good adsorption performance for both cytokines and endotoxins. Subsequently, the cationic ligand is formulated into a cationic ligand solution, which can enable the cationic ligand and the macroporous adsorption resin containing epoxy groups to fully react, and the cationic ligand and the macroporous adsorption resin containing epoxy groups first undergo a first-stage reaction at a first preset temperature, and then the temperature is raised to a second preset temperature for a second-stage reaction. Through the staged temperature increase reaction, the amino group in the cationic ligand can gradually react with the epoxy group, so that the cationic ligand is fixed on the macroporous adsorption resin, which not only improves the connection tightness between the cationic ligand and the macroporous adsorption resin, but also improves the grafting rate of the cationic ligand. After the cationic ligand is grafted onto the skeleton of the macroporous adsorption resin, a secondary amine structure is formed, which has a strong electrostatic adsorption effect in the solution and can form a strong electrostatic force with the phosphate group on the endotoxin lipid A, and adsorb endotoxin through electrostatic action. At the same time The hydrophobic fatty chains in the cationic ligand can be adsorbed together with the long fatty chains of the lipid A part of the endotoxin through hydrophobic interaction. Through the form of dual adsorption sites of electrostatic adsorption and hydrophobic adsorption, the endotoxin can be captured or anchored, thereby improving the adsorption capacity of the adsorption resin used for blood perfusion for endotoxin; in addition, after the macroporous adsorption resin is grafted with the cationic ligand, the active hydrophobic fatty chains are distributed on the outside of the macroporous adsorption resin and the inner surface of the pores, which can improve the hydrophobic properties of the macroporous adsorption resin. The hydrophobic fatty chains on the cationic ligand can move lipophilic and hydrophobic macromolecules such as cytokines toward the macroporous adsorption resin through lipophilic and hydrophobic interactions, and then through the combination of the hydrophobic fatty chains on the cationic ligand and the hydrophobic interaction of the macroporous adsorption resin itself, the adsorption of lipophilic and hydrophobic macromolecules such as cytokines and endotoxins containing hydrophobic lipid A can be greatly improved.

[0044] Compared to existing preparation methods involving modified grafting on styrene resin microspheres, the preparation method for the adsorption resin for hemoperfusion provided in this embodiment is simple and efficient, with mild reaction conditions, suitable for industrial production. It requires only two steps to complete the preparation of a cross-linked adsorption resin, eliminating the need for multi-step intermediate grafting processes to modify the adsorption resin and the need for further cross-linking to further improve the performance of the adsorption resin. This avoids the introduction of new initiators or catalysts that can lead to excessive residues or excessive side reaction products. This preparation method is specifically designed based on the molecular structural characteristics of endotoxins, significantly improving the adsorption capacity for endotoxins, and ensuring that the prepared adsorption resin has good adsorption effects on both endotoxins and cytokines.

[0045] The second aspect of the embodiment of the present application provides an adsorption resin for blood perfusion, which is prepared using the preparation method described in the first aspect. The adsorption resin provided in this embodiment can simultaneously reduce the levels of endotoxins and cytokines in the blood.

[0046] A third aspect of an embodiment of the present application provides an application of an adsorption resin for blood perfusion. The adsorption resin can be used in a blood purification device, which includes a blood perfusion device. The adsorption resin can be filled in the blood perfusion device.

[0047] Combine Figure 2 As shown, this embodiment provides a hemoperfusion device, which includes a cartridge 10 and end caps 20. The cartridge 10 has openings at both ends along the axial direction. The cartridge 10 is hollow and filled with a storage fluid and an adsorption resin 30. The end caps 20 are used to seal the openings of the cartridge 10. Two end caps 20 are respectively installed at the two axial openings of the cartridge 10. The end caps 20 include end caps and locking nuts. The end caps are used to seal the openings of the cartridge 10 and are connected to the cartridge 10 via the locking nuts. The end cap 20 also includes a cap, a grid and a sealing ring, wherein the end cap is provided with a blood nozzle and a cover body protruding from the cylinder 10 in the axial direction, the cover body is arranged around the blood nozzle, the cover body and the blood nozzle are integrally formed, and the blood nozzle is connected to the inner ring space of the cover body, the cover body extends into the inner cavity of the cylinder 10, and part of the outer peripheral wall of the cap body contacts part of the inner cylinder wall of the cylinder 10, wherein the grid is arranged between the cover body and the end of the cylinder 10, and a filter is arranged on the grid, and the filter covers the end opening of the cylinder 10, and the sealing ring is arranged in the space enclosed by the outer peripheral wall of the cover body, the grid and the inner cylinder wall of the cylinder 10 to ensure the sealing of the end cap, and the cap is used to cover the blood nozzle to achieve sealing of the blood nozzle.

[0048] The adsorption resin provided in this embodiment can be used in a blood purification device and filled in a blood perfusion device. The use of a blood perfusion device product containing the adsorption resin has important clinical significance for the treatment of acute and critical diseases such as sepsis, endotoxemia, septic shock, multiple organ dysfunction syndrome (MODS), severe trauma, liver disease, acute pancreatitis, etc., thereby providing a more comprehensive and effective treatment effect.

[0049] In order to further explain the present invention in detail, the present invention will be further described below with reference to specific examples. Unless otherwise specified, the experimental methods used in the examples of the present invention are conventional methods; unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention are all purchased from the market.

[0050] Example 1

[0051] This embodiment provides an adsorption resin for blood perfusion, comprising the following steps:

[0052] (1) Preparation of macroporous adsorption resin containing epoxy groups:

[0053] In a three-necked flask, 1 L of a 0.2% by mass methylcellulose aqueous solution was prepared as an aqueous phase solution. 15 g of monomer styrene, 5 g of allyl glycidyl ether, 80 g of a cross-linking agent divinylbenzene, 80 g of a porogen toluene, 80 g of isooctyl alcohol, and 1 g of an initiator tert-butyl peroxy-2-hexylhexanoate were mixed uniformly to form a mixed oil phase. The mixed oil phase was added to the aqueous phase solution. Under mechanical stirring, the droplet size was controlled, the temperature was raised to 75° C. and the reaction was continued for 5 h. The temperature was then raised to 85° C. and the reaction was continued for 3 h to fully solidify the resin. The resin was washed with water and sieved to obtain a macroporous adsorption resin with epoxy groups uniformly embedded in the resin skeleton. The specific surface area of the macroporous adsorption resin containing epoxy groups (measured by the BET method) was 643 m 2 / g, the average pore diameter is 12.7nm, and the content of epoxy groups is 59μmol / mL.

[0054] (2) Preparation of adsorption resin for blood perfusion:

[0055] 100 mL of the macroporous adsorption resin containing epoxy groups prepared in step (1) was measured and added to 100 mL of an ethanol aqueous solution (50% ethanol) containing 10% (mass fraction) hexylamine. 2 g of sodium hydroxide was added and stirred evenly. The mixture was heated to 60° C. and reacted for 10 h. Stirring was continued during the reaction. The mixture was then heated to 70° C. and reacted for 4 h. After the reaction, the mixture was rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. The mixture was then washed with a large amount of ethanol to remove organic residues from the macroporous adsorption resin. A macroporous adsorption resin modified with a cationic ligand grafted thereon was obtained, i.e., an adsorption resin for hemoperfusion. The immobilized hexylamine content on the adsorption resin for hemoperfusion was determined to be 51 μmol / mL.

[0056] Example 2

[0057] This embodiment provides an adsorption resin for blood perfusion, comprising the following steps:

[0058] (1) Preparation of macroporous adsorption resin containing epoxy groups:

[0059] In a three-necked flask, 1 L of a 0.5% by mass methyl cellulose aqueous solution was prepared as an aqueous phase solution. 10 g of a monomer, glycidyl methacrylate, 90 g of a cross-linking agent, 60 g of toluene, 20 g of n-heptane, 80 g of methyl isobutyl carbinol, and 1 g of an initiator, tert-butyl peroxy-2-hexylhexanoate, were mixed uniformly to form a mixed oil phase. The mixed oil phase was added to the aqueous phase solution. Under mechanical stirring, the droplet size was controlled, the temperature was raised to 75° C. and the reaction was continued for 5 h. The temperature was then raised to 85° C. and the reaction was continued for 3 h to fully solidify the resin. The resin was washed with water and sieved to obtain a macroporous adsorption resin having epoxy groups uniformly embedded in the resin skeleton. The specific surface area (measured by the BET method) of the macroporous adsorption resin containing epoxy groups was 702 m 2 / g, the average pore diameter is 12.1nm, and the content of epoxy groups is 105μmol / mL.

[0060] (2) Preparation of adsorption resin for blood perfusion:

[0061] 100 mL of the macroporous adsorption resin containing epoxy groups prepared in step (1) was measured and added to 100 mL of an ethanol aqueous solution (50% ethanol) containing 12% (mass fraction) hexadecylamine. 2 g of sodium hydroxide was added and stirred evenly. The mixture was heated to 55° C. and reacted for 8 h. Stirring was continued during the reaction. The mixture was then heated to 65° C. and reacted for 8 h. After the reaction was completed, the mixture was rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. The mixture was then washed with a large amount of ethanol to remove organic residues in the macroporous adsorption resin. A macroporous adsorption resin modified by grafting of cationic ligands was obtained, i.e., an adsorption resin for blood perfusion. The immobilized amount of hexadecylamine on the adsorption resin for blood perfusion was tested to be 90 μmol / mL.

[0062] Example 3

[0063] This embodiment provides an adsorption resin for blood perfusion, comprising the following steps:

[0064] (1) Preparation of macroporous adsorption resin containing epoxy groups: This step is the same as step (1) in Example 2, in which a macroporous adsorption resin containing epoxy groups is prepared.

[0065] (2) Preparation of adsorption resin for blood perfusion:

[0066] 100 mL of the macroporous adsorption resin containing epoxy groups prepared in step (1) was measured and added to 100 mL of an ethanol aqueous solution (50% ethanol) containing 12% (mass fraction) hexadecylamine. 2 g of sodium hydroxide was added and stirred evenly. The temperature was raised to 55° C. and the reaction was continued for 8 h. 1 g of serine was added and stirred evenly. The reaction was continued at 65° C. for 8 h. Stirring was continued throughout the reaction process. After the reaction was completed, the mixture was rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. The mixture was then washed with a large amount of ethanol to remove organic residues in the macroporous adsorption resin. A macroporous adsorption resin modified with cationic ligand grafting was obtained, i.e., an adsorption resin for blood perfusion. The total amount of hexadecylamine and serine immobilized on the adsorption resin for blood perfusion was tested to be 93 μmol / mL.

[0067] Example 4

[0068] This embodiment provides an adsorption resin for blood perfusion, comprising the following steps:

[0069] (1) Preparation of macroporous adsorption resin containing epoxy groups: This step is the same as step (1) in Example 2, in which a macroporous adsorption resin containing epoxy groups is prepared.

[0070] (2) Preparation of adsorption resin for blood perfusion:

[0071] 100 mL of the epoxy-containing macroporous adsorption resin prepared in step (1) was measured and added to 100 mL of an aqueous solution containing 10% (mass fraction) lysine. 2 g of sodium hydroxide was added and stirred evenly. The mixture was heated to 65° C. and reacted for 16 h. Stirring was continued throughout the reaction. After the reaction, the mixture was rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. The mixture was then washed with a large amount of ethanol to remove organic residues in the macroporous adsorption resin. A cationic ligand-grafted macroporous adsorption resin, i.e., an adsorption resin for hemoperfusion, was obtained. The lysine loading on the adsorption resin for hemoperfusion was tested to be 98 μmol / mL.

[0072] Comparative Example 1

[0073] The macroporous adsorption resin containing epoxy groups prepared in step (1) of Example 1 was cleaned by washing with water and a large amount of anhydrous ethanol and used as Comparative Example 1.

[0074] Comparative Example 2

[0075] The macroporous adsorption resin containing epoxy groups prepared in step (1) of Example 2 was cleaned by washing with water and a large amount of anhydrous ethanol and used as Comparative Example 2.

[0076] Comparative Example 3

[0077] The adsorbent in the commercially available CytoSorb adsorption column was used as Comparative Example 3.

[0078] Comparative Example 4

[0079] This comparative example provides an adsorption resin for blood perfusion, comprising the following steps:

[0080] (1) Preparation of macroporous adsorption resin containing epoxy groups: This step is the same as step (1) in Example 2, in which a macroporous adsorption resin containing epoxy groups is prepared.

[0081] (2) Preparation of adsorption resin for blood perfusion:

[0082] 100 mL of the epoxy group-containing macroporous adsorption resin prepared in step (1) was measured and added to 100 mL of an aqueous solution containing 10% (mass fraction) polyethyleneimine (average molecular weight 4500), 2 g of sodium hydroxide was added and stirred evenly, and the mixture was heated to 50° C. and reacted for 16 h. Stirring was continued throughout the reaction process. After the reaction was completed, the mixture was first rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. Subsequently, the mixture was washed with a large amount of ethanol to remove organic residues in the macroporous adsorption resin. A macroporous adsorption resin modified with amino cationic ligand grafting was obtained, i.e., an adsorption resin for hemoperfusion. The immobilized polyethyleneimine content on the adsorption resin for hemoperfusion was tested to be 55 μmol / mL.

[0083] Comparative Example 5

[0084] This comparative example provides an adsorption resin for blood perfusion, comprising the following steps:

[0085] (1) Preparation of macroporous adsorption resin containing epoxy groups: This step is the same as step (1) in Example 2, in which a macroporous adsorption resin containing epoxy groups is prepared.

[0086] (2) Preparation of adsorption resin for blood perfusion:

[0087] 100 mL of the macroporous adsorption resin containing epoxy groups prepared in step (1) was measured and added to 100 mL of a solution containing 10% (mass fraction) aspartic acid. 2 g of sodium hydroxide was added and stirred evenly. The mixture was heated to 65° C. and reacted for 16 h. Stirring was continued throughout the reaction. After the reaction was completed, the mixture was rinsed with water for 2 h, then washed with 1% hydrochloric acid solution for 2 h, and then washed with water until neutral. The mixture was then washed with a large amount of ethanol to remove organic residues in the macroporous adsorption resin. A macroporous adsorption resin modified with aspartic acid ligand grafting was obtained, i.e., an adsorption resin for hemoperfusion. The immobilized amount of aspartic acid on the adsorption resin for hemoperfusion was tested to be 93 μmol / mL.

[0088] Comparative Example 6

[0089] This comparative example provides an adsorption resin for blood perfusion, comprising the following steps:

[0090] (1) Epoxidation modification of polystyrene-based macroporous resin:

[0091] In a 1000mL three-necked flask, 600mL of an aqueous solution containing 1.5wt% gelatin was added, and a mixed organic phase consisting of 40g of divinylbenzene, 10g of ethylstyrene, 25g of porogen toluene, 10g of n-heptane, 25g of n-octanol, 20g of diisobutyl ketone, and 0.5g of initiator tert-butyl peroxy-2-hexylhexanoate was added. Under mechanical stirring, the oil droplet size was controlled, and the temperature was raised to 70℃ for reaction for 2h. After sampling and observing the shape of the oil droplets, 1g of the mixture was added dropwise to the three-necked flask. 0g allyl glycidyl ether, and keep the reaction at 70℃ for 5 hours, then raise the reaction temperature to 80℃ and continue the reaction for 4 hours to fully crosslink and cure. After the reaction is completed, cool, filter out the mother liquor, extract with acetone for 12 hours, wash with water until there is no acetone smell, filter, dry, sieve, and select the resin with a particle size of 0.6mm to 1.0mm to obtain the desired macroporous resin containing epoxy groups in the outer layer skeleton. The specific surface area (BET) of the macroporous resin containing epoxy groups is 722m 2 / g, the average pore diameter is 9.2nm, and the content of grafted epoxy groups in the outer skeleton of the macroporous resin containing epoxy groups is measured to be 1.2μmol / mL.

[0092] (2) Preparation of an adsorption resin for hemoperfusion: This step was performed using the same method as step (2) in Example 2 to prepare a macroporous adsorption resin containing epoxy groups. Testing revealed that the immobilized capacity of the cationic ligand (hexadecylamine) on the adsorption resin for hemoperfusion was 1.0 μmol / mL.

[0093] Comparative Example 7

[0094] This comparative example provides a zwitterion-modified adsorption resin for blood perfusion, comprising the following steps:

[0095] (1) 100 g of divinylbenzene (DVB 80), 5 g of glycidyl methacrylate, 160 g of a porogen (80 g of toluene, 80 g of n-heptane), and 2 g of benzoyl peroxide were mixed to obtain an oil phase, wherein the mass ratio of divinylbenzene, glycidyl methacrylate, porogen, and benzoyl peroxide was 1:0.05:1.6:0.02;

[0096] (2) 400 g of water and 10 g of gelatin were mixed to obtain an aqueous phase, wherein the mass ratio of water to gelatin was 1:0.025;

[0097] (3) adding the oil phase obtained in step (1) to the water phase obtained in step (2) (the mass ratio of the oil phase to the water phase is 1:1.5), and performing suspension polymerization at 80° C. for 12 h. After the reaction is completed, the mixture is stirred and washed with a 95% by volume ethanol solution five times for 3 h each time, and then dried at 70° C. to obtain a cross-linked porous resin containing epoxy groups;

[0098] (4) 100 g of the epoxy group-containing cross-linked porous resin prepared in step (3) was mixed with 400 g of water, 10 g of N, N-dimethyl-1, 3-propylenediamine was added, and a ring-opening addition reaction was carried out at 80° C. for 1 hour. After the reaction was completed, the mixture was stirred and washed with pure water for 3 times, each time for 3 hours, and then stirred and washed with 100% volume concentration ethanol for 3 times, each time for 3 hours, to obtain a cross-linked porous resin containing tertiary amine groups, wherein the mass ratio of the epoxy group-containing cross-linked porous resin, water and N, N-dimethyl-1, 3-propylenediamine was 1:4:0.1;

[0099] (5) The cross-linked porous resin containing tertiary amine groups obtained in step (4) was mixed with 600 g of solvent (300 g of ethanol and 300 g of acetone), 12 g of 1,3-propane sultone was added, and a quaternization reaction was carried out at 60° C. for 12 h. The mixture was stirred and washed 5 times with a 75% volume concentration ethanol solution for 3 h each time, and then dried at 80° C. to obtain a cross-linked zwitterion modified adsorbent, wherein the mass ratio of the cross-linked porous resin containing tertiary amine groups, the solvent and the 1,3-propane sultone was 1:6:0.12.

[0100] Test Example 1

[0101] The adsorption performance of the adsorption resins in the examples and comparative examples on cytokines IL-6, TNF-α and endotoxin was tested. The specific test methods are as follows:

[0102] The adsorption resins in Examples 1 to 4 and Comparative Examples 1 to 7 were used as adsorbents, and 1 mL of each group of adsorbents was accurately measured and added to 10 mL of plasma containing 800 pg / mL IL-6, 800 pg / mL TNF-α, 5 μg / mL β2 microglobulin, and 2 EU / mL endotoxin. After shaking at 37° C. for 2 hours, the adsorption was completed, and each plasma was taken for detection. The concentrations of IL-6, TNF-α, and β2 microglobulin were tested by chemiluminescence, and the content of endotoxin was tested by Limulus amebocyte lysate. The adsorption performance results of the adsorbents in the examples and the comparative examples for cytokines IL-6, TNF-α, β2 microglobulin, and endotoxin are shown in Table 1.

[0103] Table 1

[0104]

[0105] As can be seen from Table 1, the adsorption resins in Examples 1 to 4 all have good adsorption effects on endotoxins, and also have good adsorption effects on IL-6, TNF-α and β2 microglobulin. The IL-6 adsorption rate and the endotoxin adsorption rate can reach more than 85%, the TNF-α adsorption rate can reach more than 58%, and the β2 microglobulin adsorption rate is significantly improved, up to 95%. This shows that in this embodiment, by specifically grafting cationic ligands on the macroporous adsorption resin containing epoxy groups, the primary amine on the cationic ligand reacts with the epoxy group to form a secondary amine structure, and a hydrophobic fatty chain is introduced to achieve a dual adsorption effect of efficient electrostatic adsorption and hydrophobic adsorption of endotoxin molecules, and the hydrophobic fatty chain is also beneficial to further enhance the adsorption of IL-6, TNF-α, and β2 microglobulin.

[0106] Compared to the macroporous adsorption resin containing epoxy groups without cationic ligand grafted in Comparative Examples 1 and 2, and the macroporous adsorption resin containing epoxy groups modified with polyethyleneimine in Comparative Example 4, the adsorption performance of the adsorption resins in Examples 1 to 4 for cytokines and endotoxins can be effectively improved. Compared to the commercially available CytoSorb adsorbent in Comparative Example 3, the adsorption resins in Examples 1 to 4 have better adsorption effects on cytokines, exhibit more excellent cytokine adsorption capacity, and significantly improve the adsorption performance for endotoxins; and compared to the adsorbent in the commercially available CytoSorb adsorption column (which is polystyrene divinylbenzene resin), the macroporous adsorption resin containing epoxy groups prepared in this example does not affect the adsorption capacity of the adsorption resin for cytokines after epoxidation modification, and can significantly improve the adsorption capacity for cytokines after grafting cationic ligands. Compared to the grafting of polyethyleneimine onto the macroporous adsorption resin containing epoxy groups in Comparative Example 4, the grafting of cationic ligands in Examples 1 to 4 can significantly improve the adsorption capacity of cytokines and endotoxins. This also illustrates that the grafting of polyamine site polymers (such as polyethyleneimine and polylysine, etc.) onto the macroporous adsorption resin containing epoxy groups not only changes the structure of the adsorption resin and blocks the pores, but also enhances the hydrophilicity of the adsorption resin, resulting in a decrease in its adsorption capacity for cytokines and a weaker adsorption capacity for endotoxins, and also poor biocompatibility. Compared to Comparative Example 5, Example 4 can avoid affecting the adsorption effect of endotoxins while improving the biocompatibility of the adsorption resin by grafting lysine, while in Comparative Example 5, by grafting aspartic acid, a negatively charged and fat-free long-chain ligand, the synergistic effect of electrostatic adsorption and hydrophobic adsorption of the adsorption resin cannot be achieved, which is not conducive to further improving the adsorption performance of cytokines and endotoxins. Furthermore, Comparative Examples 4 and 5 also demonstrate that single grafted amino acids or ligands without hydrophobic fatty chains are not conducive to the adsorption of macromolecular toxins, particularly β2 microglobulin and endotoxins. Comparative Example 6 illustrates a conventional method of grafting ligands after resin surface modification. This preparation method suffers from poor grafting stability and difficulty, and can only modify epoxy groups on the adsorption resin surface. The amount of epoxy groups is small and difficult to control, resulting in a final grafted cationic ligand loading of only 1.0 μmol / mL. While this adsorption resin exhibits a certain adsorption effect on cytokines and endotoxins, its adsorption performance is far inferior to that of the adsorption resins described in Examples 1 to 4.Comparative example 7 is quaternary ammonium salt modified resin after existing epoxidation, its first preparation has porous structure and hydrophobic skeleton, then carry out quaternization reaction, although this skeleton with porous structure and hydrophobic has certain adsorption effect to cytokine, but its adsorption capacity to endotoxin is very weak, after being modified by quaternary ammonium salt, the adsorption capacity to endotoxin can be improved to a certain extent, but its hydrophilicity significantly promotes, hydrophobic interaction weakens, and electrostatic force is also weaker, resulting in the comprehensive decline of the adsorption performance of the target substances such as cytokine, endotoxin adsorbed by lipophilic hydrophobic interaction, making the adsorption capacity of the adsorbent resin finally obtained to cytokine and endotoxin weaker.And adopt the adsorption resin in the present embodiment 1 to embodiment 4, not only improve the adsorption effect to IL-6, TNF-α, also there is excellent β2 microglobulin and endotoxin adsorption capacity, realize the comprehensive promotion of adsorption performance.

[0107] Test Example 2

[0108] The hemolysis rate of the adsorption resin in each embodiment and comparative example was tested. The specific testing method is as follows:

[0109] The adsorbent resins described in Examples 1 to 4 and Comparative Examples 1 to 7 were used as adsorbents. After being moistened, the adsorbents were added with saline solution, allowed to stand at room temperature for 30 seconds, and then removed. The saline solution was then washed five times with saline solution, and then removed. 2.5 g of each adsorbent was placed in a test tube and 5 ml of saline solution was added to form the sample group. A negative control group received no adsorbent but only 5 ml of saline solution, while a positive control group received no adsorbent but only 5 ml of water for injection. The sample group, negative control group, and positive control group were all placed in a constant temperature water bath at (37±1)°C for 30 minutes. Then, 0.1 ml of diluted blood (4 ml rabbit blood + 5 ml of saline solution) was added, mixed, and placed in a (37±1)°C water bath for an additional 60 minutes, mixing every 0.5 hours. The liquid in the tube was removed and centrifuged at 800 rcf for 15 minutes. The supernatant was transferred to a cuvette, and the absorbance was measured at 545 nm using a spectrophotometer. Hemolysis rate = (AB) / (CB) x 100%, where A is the absorbance of the sample group; B is the absorbance of the negative control group; and C is the absorbance of the positive control group. The test results of the examples and comparative examples are shown in Table 2.

[0110] Test Example 3

[0111] The platelet reduction rate of the adsorption resin in each embodiment and comparative example was tested. The specific testing method is as follows:

[0112] The adsorption resins in Examples 1 to 4 and Comparative Examples 1 to 7 were used as adsorbents, and were wetted and then added with physiological saline. After standing at room temperature for 30 seconds, the mixture was removed and transferred to a polypropylene test tube and weighed. In both the Examples and Comparative Examples, an appropriate amount of blood was added at a ratio of 1 mL of blood per 0.2 g of adsorbent. The blank control group was treated with only blood without adsorbent. The test tubes were then placed in a shaking water tank at 37 ± 1 ° C with an oscillation frequency of 60 rpm. After a water bath of 60 ± 5 minutes, EDTA was added to all test tubes to terminate the reaction. 0.01 mL of 500 mM EDTA was added to each 1 mL of blood. The test tubes were then rolled and inverted at least 8 times to mix the blood samples evenly, and the blood was then transferred to a new test tube. This operation was completed in an ice water bath. Platelet count was then tested on a blood analyzer. The percentage of platelet count reduction of the adsorbent in each Example and Comparative Example compared to the blank control group was calculated. The test results of each Example and Comparative Example are shown in Table 2.

[0113] Table 2

[0114]

[0115] As can be seen from Table 2, compared to the adsorption resins of Comparative Examples 1 to 7, the adsorption resins of Examples 1 to 4 all had lower hemolysis rates, indicating that the hemolysis rates of the adsorption resins in these examples decreased to some extent. In particular, the hemolysis rate in Example 3 decreased significantly after the serine grafting. Compared to the adsorption resins of Comparative Examples 1 to 4, the adsorption resins of Examples 1 to 4 all had lower platelet reduction rates, with the platelet reduction rate being reduced by more than half. The hemolysis and platelet reduction data indicate that the adsorption resins of Examples 1 to 4 can improve their biocompatibility to a certain extent after the cationic ligand grafting, especially after the addition of a small amount of amino acids, which further improves their biocompatibility. Furthermore, although the platelet reduction rates of Examples 1 and 2 were not as low as those of Comparative Example 5, the adsorption resins of Examples 1 and 2 exhibited good cytokine and endotoxin adsorption performance and good biocompatibility. Although Comparative Examples 6 and 7 have good hemolysis rates and platelet reduction rates and good biocompatibility, the adsorption resins in Examples 1 to 4 have good biocompatibility and also good cytokine and endotoxin adsorption performance.

[0116] The adsorption resin prepared in this embodiment can simultaneously reduce the levels of endotoxins and cytokines represented by IL-6 and medium and large molecular toxins in the blood, and it also has good biocompatibility. The use of this adsorption resin for blood perfusion has important clinical significance for the treatment of patients with sepsis, endotoxemia, septic shock, multiple organ dysfunction syndrome (MODS), autoimmune diseases, etc., and can provide patients with more comprehensive and effective treatment.

[0117] Although the present disclosure is disclosed as above, the protection scope of the present disclosure is not limited thereto. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present disclosure, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. A blood purification device, characterized in that: The invention comprises a blood perfusion device, wherein the blood perfusion device is filled with an adsorption resin, and the adsorption resin is prepared by the following method: Mixing a reactive monomer and an epoxy-containing compound, wherein the reactive monomer is a mixture of a styrene monomer and a polyvinyl crosslinking agent, or the reactive monomer is a polyvinyl crosslinking agent, and the epoxy-containing compound has a carbon-carbon double bond, and the epoxy-containing compound directly reacts with the reactive monomer via the carbon-carbon double bond to undergo a suspension polymerization reaction to prepare a macroporous adsorption resin containing an epoxy group; The cationic ligand is prepared into a cationic ligand solution, wherein the cationic ligand is a monomer having a hydrophobic fatty chain at one end and an amine group at at least one end; Under alkaline conditions, the macroporous adsorption resin containing epoxy groups and the cationic ligand solution are subjected to a first-stage reaction at a first preset temperature. After the first reaction period, the temperature is raised to a second preset temperature for a second-stage reaction. The reaction period is second, so that the amino groups in the cationic ligand react with the epoxy groups and are immobilized on the macroporous adsorption resin containing epoxy groups to obtain the adsorption resin.

2. The blood purification device according to claim 1, characterized in that The hydrophobic fatty chain in the cationic ligand contains 4 to 30 carbon atoms.

3. The blood purification device according to claim 1, characterized in that The hydrophobic fatty chain in the cationic ligand contains 5 to 16 carbon atoms.

4. The blood purification device according to claim 1, characterized in that: The cationic ligand contains an amine group at one end and a hydrophobic fatty chain monomer at the other end; Alternatively, the cationic ligand is a monomer having amino groups at both ends and a hydrophobic fatty chain at the middle end.

5. The blood purification device according to claim 1, characterized in that: The volume ratio of the cationic ligand solution to the macroporous adsorption resin containing epoxy groups is 1:3 to 5:1, and the mass fraction of the cationic ligand in the cationic ligand solution is 1% to 100%.

6. The blood purification device according to claim 1, characterized in that After the first stage reaction for the first time, and before the temperature is raised to the second preset temperature for the second stage reaction, the method further includes: adding an amino acid compound to the reaction product of the first stage and mixing uniformly to obtain a grafting mixture; Wherein, the amino acid compound is a neutral amino acid compound or a basic amino acid compound.

7. The blood purification device according to claim 6, characterized in that: The mass fraction of the amino acid compound in the graft mixture does not exceed 5%.

8. The blood purification device according to claim 1, characterized in that: The first preset temperature is 25°C to 60°C, and the first time length is 1 hour to 24 hours; the second preset temperature is 60°C to 85°C, and the second time length is 1 hour to 24 hours.

9. The blood purification device according to any one of claims 1 to 3, characterized in that: The epoxy group-containing compound directly undergoes a suspension polymerization reaction with the reactive monomer via a carbon-carbon double bond to prepare a macroporous adsorption resin containing an epoxy group, comprising: The reaction monomer and the epoxy-containing compound are subjected to a suspension polymerization reaction in a dispersion medium under the action of a porogen and an initiator. The temperature of the suspension polymerization reaction is 70° C. to 90° C. After the suspension polymerization reaction is completed, a macroporous adsorption resin containing epoxy groups is obtained; the content of the epoxy groups on the macroporous adsorption resin containing epoxy groups is greater than 5 μmol / mL resin.

10. The blood purification device according to claim 1, characterized in that: Calculated on the basis of amine groups, the immobilized amount of the cationic ligand on each 1 mL of the adsorption resin is greater than 5 μmol.

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