Preparation method of hypercrosslinked macroporous adsorption resin and application of hypercrosslinked macroporous adsorption resin in blood purification products

Abstract

The invention discloses a preparation method of a hyper-crosslinked macroporous adsorption resin capable of simultaneously adsorbing bilirubin, bile acid and cytokines in blood purification. The preparation method comprises the following steps: preparing macroporous styrene-divinylbenzene resin with low crosslinking degree from a styrene monomer and a polyethylene phenyl crosslinking agent; then carrying out secondary crosslinking on macroporous styrene-divinylbenzene resin with low crosslinking degree by utilizing a Friedel-Grafts method to prepare hypercrosslinked adsorption resin, and further reacting residual chloromethyl with an amination reagent to improve the adsorption performance of bilirubin and bile acid. The prepared adsorbent can adsorb bilirubin, bile acid and cell factors at the same time, the adsorption rate of the adsorbent to bilirubin is superior to that of an anion resin bilirubin perfusion device used clinically, in combined artificial liver treatment, one perfusion device can replace two perfusion devices, the treatment cost and the operation risk are greatly reduced, and the treatment effect is good. Meanwhile, the biocompatibility is superior to that of anion resin, and the good market application value is achieved.

Description

Technical Field

[0001] This invention relates to the field of biomedical engineering technology, specifically to the preparation of an ultra-high cross-linked macroporous adsorption resin and its application in blood purification products, particularly for the simultaneous adsorption of bilirubin, bile acids and cytokines in blood. Background Technology

[0002] Bilirubin is a bile pigment, a metabolic product of aging red blood cells. In the body, bilirubin generally combines with glucuronic acid to form a water-soluble complex, which is transported by the blood to the liver for metabolism and excreted through the bile ducts. Liver damage, bile duct obstruction, and other factors can cause bilirubin to accumulate in the body, leading to hyperbilirubinemia. Excessively high levels of bilirubin can also cross the blood-brain barrier and bind to nerve nuclei, interfering with the normal function of brain cells and causing irreversible neurological damage to the brain. In severe cases, it can even be life-threatening.

[0003] Meanwhile, in patients with hyperbilirubinemia, bacteria and endotoxins in the intestines can cross the epithelial barrier and enter the mesenteric lymph nodes, thereby entering the blood and organs, promoting the formation of endotoxemia, and in turn, a large number of cytokines such as TNF-α and IL-6, exacerbating the development of the disease.

[0004] Currently, the main clinical treatments for hyperbilirubinemia include phototherapy, drug therapy, plasma exchange, and blood purification. Phototherapy is primarily for patients with mild symptoms; drug therapy has limited clinical efficacy, and patients' symptoms often do not improve promptly; plasma exchange requires large amounts of fresh plasma, which is expensive and has limited availability, and also carries the risk of cross-infection; blood purification has significant advantages in clearing bilirubin, bile acids, and cytokines, and is receiving increasing clinical attention. There are currently three main clinical modalities for blood purification therapy of hyperbilirubinemia: Baxter's MARS system (USA), Fresenius' Prometheus system (Germany), and Jianfan's DPMAS system (my country). The MARS system uses one anion exchange resin perfusion unit (diaMARS IE250) and one activated carbon perfusion unit (diaMARS AC250) to adsorb bilirubin, bile acids, and cytokines from human serum albumin, respectively. The Prometheus system uses one anion exchange resin perfusion unit (Prometh02) and one macroporous adsorption resin perfusion unit (Prometh 01) to adsorb bilirubin, bile acids, and cytokines from plasma, respectively. The DPMAS system also uses one anion exchange resin perfusion unit (BS330) and one macroporous adsorption resin perfusion unit (HA330-Ⅱ) to adsorb bilirubin, bile acids, and cytokines from plasma, respectively. These mainstream clinical treatment modalities all require two perfusion units to simultaneously adsorb bilirubin, bile acids, and cytokines, significantly increasing clinical operational risks and treatment costs.

[0005] Therefore, providing an adsorbent for blood purification that can simultaneously adsorb bilirubin, bile acids, and cytokines is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0006] The main objective of this invention is to provide a method for preparing an ultra-high cross-linked macroporous adsorption resin and its application. The resin prepared by this invention can simultaneously adsorb bilirubin, bile acids and cytokines, and the adsorption effect is significant.

[0007] To achieve the above objectives, the present invention provides a method for preparing a highly cross-linked macroporous adsorption resin, specifically comprising the following steps: 1) Suspension polymerization: A dispersion system, crosslinking agent, pore-forming agent, and initiator are added to a reactor, and a low-crosslinking degree macroporous styrene-divinylbenzene resin is obtained by suspension polymerization. The monomer is a styrene monomer; The crosslinking agent is a polyvinyl group; The amount of dispersant in the dispersion system is 0.1%-2.0% of the mass of the dispersion medium; and the volume ratio of the dispersion medium to the oil phase is 0.5-5:1. The amount of the initiator is 0.5%-2.0% of the total mass of the monomer and crosslinking agent; The amount of the porogen used is 100%-300% of the weight of the monomer; The low-crosslinking macroporous styrene-divinylbenzene resin has a particle size of 0.1-1.5 cm; 2) Chloromethylation reaction: Chlorinated methyl ether was added to the macroporous styrene-divinylbenzene resin, and after standing for a certain period of time, the mixture was stirred, and a catalyst was added to carry out the reaction to prepare chlorinated spheres. The mass of the added chloromethyl ether is 3-9 times the mass of the macroporous styrene-divinylbenzene resin; the mass of the added catalyst is 0.5-1.5 times the mass of the macroporous styrene-divinylbenzene resin. Furthermore, the catalyst can be added multiple times, preferably in two separate additions; 3) Post-crosslinking reaction: A solvent was added to the chlorine spheres, and after standing and swelling, the mixture was stirred. A catalyst was then added to carry out the reaction, thereby preparing an ultra-highly cross-linked macroporous adsorption resin. The mass of the solvent added is 3-9 times the mass of the chlorine sphere, and the mass of the catalyst added is 0.1-0.5 times the mass of the chlorine sphere. 4) Amination reaction: The highly cross-linked macroporous adsorption resin was cleaned with alcohol, and the residual alcohol was cleaned with purified water. Then, purified water and sodium hydroxide solution were added to the highly cross-linked macroporous adsorption resin at a mass ratio of 1:1-5. After heating to a certain temperature, an amination reagent was added dropwise over a certain period of time. After the reaction was completed, the resin was cleaned to obtain an amino-group-containing highly cross-linked macroporous adsorption resin. The concentration of the sodium hydroxide solution is 0.3-3 mol / L; the mass of the added sodium hydroxide is 5-20% of the mass of the ultra-high cross-linked macroporous adsorption resin, and the mass of the added amination reagent is 0.3-3 times the mass of the ultra-high cross-linked macroporous adsorption resin.

[0008] The beneficial effects of adopting the above technical solution include at least the following: the ultra-high cross-linked macroporous adsorption resin prepared by this invention is used for blood purification, especially for the simultaneous adsorption of bilirubin, bile acids, and cytokines. This invention uses styrene monomers and polyvinyl groups as cross-linking agents, obtaining a low-cross-linked macroporous styrene-divinylbenzene resin through suspension polymerization, then preparing an ultra-high cross-linked macroporous adsorption resin through chlorination and Friedel-Crafts post-cross-linking, and finally preparing an ultra-high cross-linked macroporous adsorption resin with amino groups by reacting an amination reagent with residual chloromethyl groups. This invention exhibits ultra-high adsorption performance for bilirubin, bile acids, and cytokines in patients with hyperbilirubinemia, and its bilirubin adsorption rate is superior to that of clinically used anion exchange resin bilirubin perfusion devices. In combined artificial liver treatment, one perfusion device can replace two, significantly reducing treatment costs and operational risks. It also has excellent biocompatibility, superior to anion exchange resins, and possesses significant market application value.

[0009] Preferably, the dispersion medium in step 1) is water or a saline solution with a mass fraction of 10%-30%, preferably water; the dispersant is one or a mixture of gelatin, polyvinyl alcohol or cellulose, preferably gelatin; And / or, the styrene monomer is one or a mixture of several of styrene, methylstyrene, ethylstyrene and p-hydroxystyrene, preferably styrene; And / or, the polyvinyl group is one or a mixture of several of divinylbenzene, diallylbenzene, divinylbenzenetoluene, divinylxylene and trivinylbenzene, preferably divinylbenzene; And / or, the initiator is one or a mixture of several of benzoyl peroxide, tert-butyl peroxide-2-ethylhexanoate, and tert-amyl peroxide-2-ethylhexanoate, preferably benzoyl peroxide; And / or, the pore-forming agent is one or a mixture of several of the following: benzene, toluene, xylene, n-butanol, isobutanol, n-octanol, isooctanol, methyl isobutyl methanol, methyl isobutyl ketone, cyclohexane, methylcyclohexane, n-heptane, 200# gasoline, white oil, liquid paraffin, solid paraffin, butyl acetate, ethyl acetate, butyl butyrate, oligostyrene, oligoethylene glycol, oligomethyl methacrylate, oligomethyl methacrylate, and oligovinyl acetate, preferably one or a mixture of several of the following: oligomethyl methacrylate, toluene, oligostyrene, isooctanol, 200# gasoline, liquid paraffin, and ethyl acetate.

[0010] Preferably, the catalyst in step 2) is one or a mixture of zinc chloride, ferric chloride and aluminum chloride, preferably zinc chloride.

[0011] Preferably, in step 2), the settling temperature is 20-50℃ and the settling time is 3-6 hours; the reaction temperature is 30-50℃ and the reaction time is 10-80 hours.

[0012] Preferably, the solvent in step 3) is dichloroethane and / or nitrobenzene, preferably nitrobenzene; the catalyst is one or a mixture of zinc chloride, ferric chloride and aluminum chloride, preferably zinc chloride.

[0013] Preferably, in step 3), the static swelling temperature is 30-50℃ and the static swelling time is 3-6 hours; the reaction temperature is 70-130℃ and the reaction time is 6-20 hours.

[0014] Preferably, the amination reagent in step 4) is one or a mixture of several of the following: dimethylamine, dimethylamine hydrochloride, dipropylamine, dipropylamine hydrochloride, dibutylamine, dibutylamine hydrochloride, trimethylamine, trimethylamine hydrochloride, triethylamine, triethylamine hydrochloride, tripropylamine, tripropylamine hydrochloride, ethanolamine, ethanolamine hydrochloride, ethylenediamine, ethylenediamine hydrochloride, propylenediamine, propylenediamine hydrochloride, butylenediamine, and butylenediamine hydrochloride, preferably dimethylamine, ethylenediamine, or ethanolamine.

[0015] Preferably, in step 4), the specific temperature is 20-80℃, the specific time is 2-5 hours, and the reaction time is 5-50 hours.

[0016] This invention also provides the application of the above-mentioned ultra-high cross-linked macroporous adsorption resin in blood purification products.

[0017] Furthermore, the application of the ultra-highly cross-linked macroporous adsorption resin in products that simultaneously adsorb bilirubin, bile acids, and cytokines is discussed. The cytokines are preferably TNF-α and IL-6. Detailed Implementation

[0018] The technical solution of the present invention will be further described below through embodiments. Example 1

[0019] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: Suspension polymerization Add 500 mL of a 1.0 wt% gelatin aqueous solution to a 1000 mL three-necked flask, then add 21 g of diethyl ether. A mixed oil phase consisting of divinylbenzene, 79g styrene, 60g oligomeric methyl methacrylate (degree of polymerization 100-1000), 40g toluene, and 1.0g benzoyl peroxide was reacted at 78℃ for 4 hours under mechanical stirring, then heated to 83℃ for 5 hours, and then heated to 87℃ for 6 hours. After the reaction was completed, the pore-forming agent was extracted, and the mixture was washed with water until the washing liquid was clear. The mixture was then filtered, dried, and sieved. Resin with a particle size of 0.2-0.8mm was selected to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0020] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 40℃ and react for 80h; after the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0021] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0022] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of dimethylamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain ultra-high cross-linked macroporous adsorption resin with amine groups. Example 2

[0023] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of an aqueous solution containing 1.0 wt% gelatin to a 1000 mL three-necked flask, then add a mixed oil phase consisting of 21 g divinylbenzene, 79 g styrene, 60 g oligostyrene (degree of polymerization 100-1000), 40 g toluene, and 1.0 g benzoyl peroxide. Under mechanical stirring, the mixture is heated to 78 °C for 4 h, then heated to 83 °C for 5 h to cure, and then heated to 87 °C for 6 h. After the reaction is complete, the pore-forming agent is extracted, and the mixture is washed with water until the washing liquid is clear. The mixture is then filtered, dried, and sieved. Resin with a particle size of 0.2-0.8 mm is selected to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0024] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 40℃ and react for 80h. After the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0025] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0026] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of dimethylamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain ultra-high cross-linked macroporous adsorption resin with amine groups. Example 3

[0027] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of an aqueous solution containing 1.0 wt% gelatin to a 1000 mL three-necked flask, then add a mixed oil phase consisting of 21 g divinylbenzene, 79 g styrene, 125 g isooctyl alcohol, 45 g toluene, and 1.0 g benzoyl peroxide. Under mechanical stirring, heat to 78 °C for 4 h, then heat to 83 °C for 5 h, and then heat to 87 °C for 6 h. After the reaction is complete, extract the pore-forming agent, wash with water until the washing liquid is clear, filter, dry, and sieve. Select resin with a particle size of 0.2-0.8 mm to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0028] 2) Chloromethylation reaction Add 20g of the aforementioned low-crosslinking macroporous styrene-divinylbenzene resin to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, then start the stirrer and add 20g of anhydrous zinc chloride in two portions. Increase the temperature to 40℃ and react for 80h. After the reaction is complete, cool to room temperature, filter out the mother liquor, wash three times with methanol, wash with water until no methanol odor remains, filter under vacuum, and dry.

[0029] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0030] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of ethanolamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin. Example 4

[0031] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of an aqueous solution containing 1.0 wt% gelatin to a 1000 mL three-necked flask, then add a mixed oil phase consisting of 21 g divinylbenzene, 79 g styrene, 125 g 200# gasoline, 45 g toluene, and 1.0 g benzoyl peroxide. Under mechanical stirring, the mixture is heated to 78 °C for 4 h, then heated to 83 °C for 5 h to cure, and then heated to 87 °C for 6 h. After the reaction is complete, the pore-forming agent is extracted, washed with water until the washing liquid is clear, filtered, dried, and sieved. Resin with a particle size of 0.2-0.8 mm is selected to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0032] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 40℃ and react for 80h. After the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0033] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0034] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of ethanolamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin. Example 5

[0035] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of an aqueous solution containing 1.0 wt% gelatin to a 1000 mL three-necked flask, then add a mixed oil phase consisting of 21 g divinylbenzene, 79 g styrene, 125 g liquid paraffin, 45 g toluene, and 1.0 g benzoyl peroxide. Under mechanical stirring, heat to 78 °C and react for 4 h, then heat to 83 °C and cure for 5 h, then heat to 87 °C and react for 6 h. After the reaction is complete, extract the pore-forming agent, wash with water until the washing liquid is clear, filter, dry, and sieve. Select resin with a particle size of 0.2-0.8 mm to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0036] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 40℃ and react for 80h. After the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0037] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0038] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of ethylenediamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin. Example 6

[0039] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of an aqueous solution containing 1.0 wt% gelatin to a 1000 mL three-necked flask, then add a mixed oil phase consisting of 21 g divinylbenzene, 79 g styrene, 125 g ethyl acetate, 45 g toluene, and 1.0 g benzoyl peroxide. Under mechanical stirring, heat to 78 °C for 4 h, then heat to 83 °C for 5 h, and then heat to 87 °C for 6 h. After the reaction is complete, extract the pore-forming agent, wash with water until the washing liquid is clear, filter, dry, and sieve. Select resin with a particle size of 0.2-0.8 mm to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0040] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 4h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 40℃ and react for 80h. After the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0041] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 40℃ for 4h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 120℃, keep the reaction at this temperature for 12h, then cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0042] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of ethylenediamine aqueous solution dropwise over 3 hours, and react for 10 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin. Example 7

[0043] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of a 2.0 wt% gelatin aqueous solution to a 1000 mL three-necked flask, then add 21 g of diethyl ether. A mixed oil phase consisting of alkenylbenzene toluene, 79g ethylstyrene, 60g n-butanol, 40g toluene, and 1.0g tert-butyl peroxide-2-ethylhexanoate was reacted at 78℃ for 4 hours under mechanical stirring, then heated to 83℃ for 5 hours, and then heated to 87℃ for 6 hours. After the reaction was completed, the pore-forming agent was extracted, and the mixture was washed with water until the washing liquid was clear. The mixture was then filtered, dried, and sieved. Resin with a particle size of 0.1-0.9mm was selected to obtain macroporous styrene-divinylbenzene resin with low crosslinking degree.

[0044] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 30℃ for 5h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 45℃ and react for 75h; after the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0045] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let it stand at 30℃ for 6 hours to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 130℃, keep the temperature for 17 hours, cool to room temperature, filter out the mother liquor, wash with alcohol 3 times and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0046] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of dipropylamine aqueous solution dropwise over 3 hours, and react for 15 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin. Example 8

[0047] This embodiment provides a method for preparing an ultra-highly cross-linked macroporous adsorption resin, specifically including: 1) Suspension polymerization Add 500 mL of a 2.0 wt% cellulose aqueous solution to a 1000 mL three-necked flask, along with a mixed oil phase consisting of 35 g divinylxylene, 115 g p-hydroxystyrene, 100 g polyvinyl acetate (degree of polymerization 100-1000), 60 g xylene, and 1.5 g tert-amyl peroxide-2-ethylhexanoate. Under mechanical stirring, the mixture is heated to 78 °C for 4 h, then heated to 83 °C for 5 h to cure, and then heated to 87 °C for 6 h. After the reaction, the pore-forming agent is extracted, and the mixture is washed with water until the washing liquid is clear. The mixture is then filtered, dried, and sieved. Resin with a particle size of 0.4-1.2 mm is selected to obtain a low-crosslinking macroporous styrene-divinylbenzene resin.

[0048] 2) Chloromethylation reaction Add 20g of the low-crosslinking macroporous styrene-divinylbenzene resin obtained above to a 500mL three-necked flask, add 100g of chloromethyl ether, let stand at 20℃ for 5h, start the stirrer, add 20g of anhydrous zinc chloride in two portions, heat to 50℃ and react for 75h; after the reaction is completed, cool to room temperature, filter out the mother liquor, wash 3 times with methanol, wash with water until there is no methanol odor, filter under vacuum and dry.

[0049] 3) Post-crosslinking reaction Add 20g of the obtained chlorine beads to a 500mL three-necked flask, add 140g of nitrobenzene, let stand at 45℃ for 5h to swell, then start mechanical stirring, add 5g of zinc chloride, heat to 125℃, keep the temperature for 16h, cool to room temperature, filter out the mother liquor, wash 3 times with alcohol and then wash with water until there is no alcohol odor, filter by suction, and dry to obtain ultra-high cross-linked macroporous adsorption resin.

[0050] 4) Amination reaction Take 20g of ultra-high cross-linked macroporous adsorption resin, clean it with alcohol, then clean the remaining alcohol with purified water, add 60g of purified water, then add 2g of 1mol / L sodium hydroxide, heat to 45℃, add 40g of triethylamine aqueous solution dropwise over 3 hours, and react for 15 hours to obtain an amino-group ultra-high cross-linked macroporous adsorption resin.

[0051] Adsorption performance evaluation The resins obtained in Examples 1-8 were used as the same type of blood perfusion adsorbent, and the adsorbents were used in the same way. At the same time, the adsorbents in commercially available Jianfan BS adsorption columns and commercially available Boxin DX adsorption columns were used as control examples. Plasma from the same liver disease patient was used to conduct in vitro plasma adsorption experiments on the above examples and control examples respectively.

[0052] The method is as follows: 1 ml of wet resin (Examples 1-8) was measured and placed in a 50 ml stoppered conical flask. 10 ml of plasma from the aforementioned liver disease patients was added. The flask was then placed in a reciprocating constant-temperature water bath shaker at 37±1℃ and a rate of 120±10 times / min for 2 hours for adsorption. The concentration of the target substance before and after adsorption was measured and calculated. The results are shown in Table 1.

[0053] Table 1

[0054] As can be seen from Table 1, the ultra-high cross-linked macroporous adsorption resin prepared in the embodiments of the present invention has a strong adsorption performance for bilirubin and bile acids, which is superior to the anion exchange resin bilirubin perfusion device used in clinical practice, and it also has a very high adsorption rate for cytokines represented by TNF-α and IL-6.

[0055] The above description is merely a preferred embodiment of the present invention and does not constitute any limitation on the technical scope of the present invention. Therefore, any minor modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention shall still fall within the scope of the technical solution of the present invention.

Claims

1. A method for preparing an ultra-highly cross-linked macroporous adsorption resin, characterized in that, Specifically, the steps include the following: 1) Suspension polymerization: A dispersion system, monomer, crosslinking agent, pore-forming agent and initiator are added to a reactor, and a low-crosslinking degree macroporous styrene-divinylbenzene resin is obtained by suspension polymerization. The monomer is a styrene-based monomer. The crosslinking agent is a polyvinyl group; The amount of dispersant in the dispersion system is 0.1%-2.0% of the mass of the dispersion medium; and the volume ratio of the dispersion medium to the oil phase is 0.5-5:

1. The amount of the initiator is 0.5%-2.0% of the total mass of the monomer and crosslinking agent; The amount of the porogen used is 100%-300% of the weight of the monomer; The low-crosslinking macroporous styrene-divinylbenzene resin has a particle size of 0.1-1.5 cm; Chloromethylation reaction: Chloromethyl ether was added to the macroporous styrene-divinylbenzene resin, and after standing for a certain period of time, the mixture was stirred, and a catalyst was added to carry out the reaction to prepare chlorine spheres. The mass of the added chloromethyl ether is 3-9 times the mass of the macroporous styrene-divinylbenzene resin; the mass of the added catalyst is 0.5-1.5 times the mass of the macroporous styrene-divinylbenzene resin. 3) Post-crosslinking reaction: A solvent was added to the chlorine spheres, and after standing and swelling, the mixture was stirred. A catalyst was then added to carry out the reaction, thereby preparing an ultra-highly cross-linked macroporous adsorption resin. The mass of the solvent added is 3-9 times the mass of the chlorine sphere, and the mass of the catalyst added is 0.1-0.5 times the mass of the chlorine sphere. 4) Amination reaction: The highly cross-linked macroporous adsorption resin was cleaned with alcohol, and the residual alcohol was cleaned with purified water. Then, purified water and sodium hydroxide solution were added to the highly cross-linked macroporous adsorption resin at a mass ratio of 1:1-5. After heating to a certain temperature, an amination reagent was added dropwise over a certain period of time. After the reaction was completed, the resin was cleaned to obtain an amino-group-containing highly cross-linked macroporous adsorption resin. The concentration of the sodium hydroxide solution is 0.3-3 mol / L; the mass of the added sodium hydroxide is 5-20% of the mass of the ultra-high cross-linked macroporous adsorption resin, and the mass of the added amination reagent is 0.3-3 times the mass of the ultra-high cross-linked macroporous adsorption resin.

2. The method for preparing a highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 1) The dispersion medium is water or a saline solution with a mass fraction of 10%-30%; the dispersant is one or a mixture of gelatin, polyvinyl alcohol, or cellulose. And / or, the styrene monomer is one or a mixture of several of styrene, methylstyrene, ethylstyrene and p-hydroxystyrene; And / or, the polyvinyl group is one or a mixture of several of divinylbenzene, diallylbenzene, divinylbenzenetoluene, divinylxylene, and trivinylbenzene; And / or, the initiator is one or a mixture of several of benzoyl peroxide, tert-butyl peroxide-2-ethylhexanoate, and tert-amyl peroxide-2-ethylhexanoate; And / or, the pore-forming agent is one or a mixture of several of the following: benzene, toluene, xylene, n-butanol, isobutanol, n-octanol, isooctanol, methyl isobutyl methanol, methyl isobutyl ketone, cyclohexane, methylcyclohexane, n-heptane, 200# gasoline, white oil, liquid paraffin, solid paraffin, butyl acetate, ethyl acetate, butyl butyrate, oligostyrene, oligoethylene glycol, oligomethyl acrylate, oligomethyl methacrylate, and oligovinyl acetate.

3. The method for preparing a highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 2) The catalyst is one or a mixture of zinc chloride, ferric chloride and aluminum chloride.

4. The method for preparing a highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 2) The settling temperature is 20-50℃ and the settling time is 3-6 hours; the reaction temperature is 30-50℃ and the reaction time is 10-80 hours.

5. The method for preparing an ultra-highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 3) The solvent is dichloroethane and / or nitrobenzene; the catalyst is one or a mixture of zinc chloride, ferric chloride and aluminum chloride.

6. The method for preparing an ultra-highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 3) The static swelling temperature is 30-50℃, and the static swelling time is 3-6 hours; the reaction temperature is 70-130℃, and the reaction time is 6-20 hours.

7. The method for preparing an ultra-highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 4) The amination reagent is one or a mixture of several of the following: dimethylamine, dimethylamine hydrochloride, dipropylamine, dipropylamine hydrochloride, dibutylamine, dibutylamine hydrochloride, trimethylamine, trimethylamine hydrochloride, triethylamine, triethylamine hydrochloride, tripropylamine, tripropylamine hydrochloride, ethanolamine, ethanolamine hydrochloride, ethylenediamine, ethylenediamine hydrochloride, propylenediamine, propylenediamine hydrochloride, butylenediamine, and butylenediamine hydrochloride.

8. The method for preparing a highly cross-linked macroporous adsorption resin according to claim 1, characterized in that, Step 4) The specified temperature is 20-80℃, the specified time is 2-5 hours, and the reaction time is 5-50 hours.

9. The application of the ultra-high cross-linked macroporous adsorption resin obtained by any one of the preparation methods according to claims 1-8 in blood purification products.

10. The application of the ultra-high cross-linked macroporous adsorption resin according to claim 9 in blood purification products, characterized in that, Application of the ultra-highly cross-linked macroporous adsorption resin in products that simultaneously adsorb bilirubin, bile acids, and cytokines. The cytokines are preferably TNF-α and IL-6.

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